Lactobacillus acidophilus for relieving chemotherapy-induced diarrhea and improving constipation and application thereof

Abstract

The application discloses a lactobacillus acidophilus for relieving chemotherapy-induced diarrhea and improving constipation and application thereof, and relates to the technical field of microorganisms. Lactobacillus acidophilus The lactobacillus acidophilus LIHUO 1978 has good safety, in-vitro probiotic characteristics and in-vitro immune regulation characteristics, and has the effects of relieving chemotherapy-induced diarrhea, improving constipation and regulating intestinal flora, and has a good application prospect in preparation of medicine for relieving chemotherapy-induced diarrhea and / or preparation of products for improving constipation and regulating intestinal flora.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, specifically to a strain of Lactobacillus acidophilus that relieves chemotherapy-induced diarrhea and improves constipation, and its applications. Background Technology

[0002] Chemotherapy remains the primary treatment strategy for most cancer patients, especially those with advanced cancer, distant metastases, and resistance to molecular targeted therapy or immunotherapy. Gastrointestinal toxicity is a common complication induced by chemotherapy, including clinical symptoms such as diarrhea, constipation, nausea, and vomiting. Chemotherapy-induced diarrhea, caused by chemotherapy, severely impacts patients' quality of life. The inducing factors for chemotherapy-induced diarrhea include intestinal epithelial damage, inflammation leading to impaired absorption and secretion in the small intestine, and disruption of the mucosal barrier. Opioid derivatives such as loperamide, deodorant opiate, and octreotide are commonly used to treat chemotherapy-induced diarrhea, but these drugs are often accompanied by numerous side effects such as abdominal pain, rash, goiter, and allergies. Therefore, the development of drugs with good efficacy and high safety for the prevention or treatment of chemotherapy-induced diarrhea is urgently needed.

[0003] Constipation is more common in women than men, and more common in the elderly than in younger people. There are many causes of constipation, including intestinal obstruction, lack of exercise, low fiber intake, or personal factors. Functional constipation is the most common type, and gut microbiota imbalance is a potential trigger for functional constipation, further restricting intestinal motility and immune barrier function. Functional constipation leads to decreased productivity and affects patients' quality of life. Typical methods for preventing or treating constipation include improving diet, lifestyle modifications, and using various types of medications such as bulking agents, osmotic agents, and stimulant laxatives. These treatments have limitations, such as inconsistent efficacy, varying degrees of symptom relief, and potential safety issues. Therefore, there is an urgent need to develop effective and safe medications or dietary supplements for relieving or treating constipation.

[0004] In recent years, the effects of probiotics in regulating gut microbiota, relieving chemotherapy-induced diarrhea, and improving constipation have been confirmed, and they have a long history of human use and are relatively safe. Some probiotics, such as Lactobacillus, Bifidobacterium, and yeast, can prevent or improve chemotherapy-induced diarrhea. The potential mechanisms by which probiotics improve chemotherapy-induced diarrhea include regulating gut microbiota, regulating immunity, and reducing the levels of pro-inflammatory cytokines (Daylia T, Nutthada A, Tippawan S. Effects of Probiotics on Chemotherapy-induced Diarrhea. [J]. Nutrition and Cancer, 2023, 75 (10): 11-11.). Probiotics such as Lactobacillus can relieve constipation by regulating gut microbiota, short-chain fatty acid levels, aquaporin expression levels, neurotransmitter and hormone levels, inflammation levels, and antioxidant levels (Dimidi E, Scott MS, Whelan K. Probiotics and constipation: mechanisms of action, evidence for effectiveness and utilisation by patients and healthcare professionals [J]. Proceedings of the Nutrition Society, 2020, 79 (1): 147-157.).

[0005] The effects of probiotics in relieving chemotherapy-induced diarrhea and improving constipation still exhibit interspecies and interstrain differences (Linlin W, Lujun H, Qi X, et al. Bifidobacteria exert species-specific effects on constipation in BALB / c mice. [J]. Food & function, 2017, 8 (10): 3587-3600.). Therefore, providing a strain of Lactobacillus acidophilus with good safety, in vitro probiotic properties, in vitro immunomodulatory properties, and the ability to relieve chemotherapy-induced diarrhea, improve constipation, and regulate intestinal flora, and its application, is of great value and has considerable application prospects. Summary of the Invention

[0006] The purpose of this invention is to provide a strain of Lactobacillus acidophilus that relieves chemotherapy-induced diarrhea and improves constipation, and its applications. This strain has good safety, in vitro probiotic properties, and in vitro immunomodulatory properties. This strain can relieve chemotherapy-induced diarrhea, improve constipation, and regulate intestinal flora. This strain can be used to prepare drugs for relieving chemotherapy-induced diarrhea, and / or to prepare health foods, animal nutrition products, and drugs for improving constipation and / or regulating intestinal flora, and has good application prospects.

[0007] To achieve the above-mentioned objectives, the technical solution of the present invention is as follows:

[0008] On the one hand, the present invention provides a strain of Lactobacillus acidophilus ( Lactobacillus acidophilus ) LIHUO1978, the preservation number of the Lactobacillus acidophilus LIHUO 1978 is GDMCC No:65514.

[0009] Specifically, the 16S rDNA sequence of Lactobacillus acidophilus LIHUO 1978 is shown in SEQ ID NO:1.

[0010] Furthermore, the *Lactobacillus acidophilus* LIHUO 1978 was isolated from farm-grown light cream in Bayannur City, Inner Mongolia Autonomous Region. The colony morphology of this strain on MRS medium is white, round, moist, opaque, and with neat edges. The bacterial cell morphology is rod-shaped, arranged singly or in pairs. This strain is Gram-positive. This strain exhibits good safety: hemolysis reaction is negative, putrescine, cadaverine, histamine, and tyramine were not detected, and it is non-pathogenic. This strain has excellent in vitro probiotic properties: good growth characteristics, acid production characteristics, skim milk fermentation characteristics, and tolerance in simulated gastrointestinal fluids. This strain has good in vitro immunomodulatory properties: live bacteria of this strain can significantly promote the production of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) by normal macrophages; heat-inactivated bacteria of this strain can significantly promote the production of TNF-α and IL-10 by normal and inflammatory macrophages, and can significantly promote the production of TNF-α in a Caco-2 cell-macrophage co-culture system.

[0011] In another aspect, the present invention provides a preparation of Lactobacillus acidophilus LIHUO 1978, which is prepared by culturing Lactobacillus acidophilus LIHUO 1978 in a culture medium.

[0012] Specifically, the Lactobacillus acidophilus LIHUO 1978 preparation includes any one or more of the following: live bacteria, fermentation broth, fermentation broth precipitate, heat-inactivated bacterial cells, and lyophilized powder.

[0013] Furthermore, the fermentation broth refers to the liquid obtained after inoculating the microbial strain into a culture medium and culturing it.

[0014] Furthermore, the fermentation broth precipitation refers to the liquid precipitate after centrifugation, including free proteins, residual bacterial cells, broken cells, and culture medium residues.

[0015] Furthermore, the method for preparing the heat-inactivated bacterial cells is to sterilize the test bacterial solution of Lactobacillus acidophilus LIHUO 1978 at 121°C for 15 minutes to prepare heat-inactivated bacterial cells.

[0016] Furthermore, the method for preparing the freeze-dried powder can employ methods currently used in the art, or other methods that may emerge in the future to obtain the product in solid form of microorganisms or microbial cultures. It should be clarified that the method for preparing the freeze-dried powder should not be a condition limiting the scope of protection of this invention.

[0017] Furthermore, the culture medium can be one or more of a suitable solid culture medium, semi-solid culture medium, or liquid culture medium.

[0018] Furthermore, the culture media include, but are not limited to, MRS medium, M17 medium, RCM medium, MC medium, TTB medium, BS medium, SC medium, VRBA medium, and Bengal red medium.

[0019] In another aspect, the present invention provides a microbial agent comprising the above-mentioned Lactobacillus acidophilus LIHUO 1978 or the above-mentioned Lactobacillus acidophilus LIHUO 1978 preparation.

[0020] Specifically, the bacterial agent is a powder or liquid preparation.

[0021] Specifically, the microbial agent may be a solid, liquid, semi-solid, or any physical form containing the necessary active ingredients to achieve any of the applications, functions, or effects described in this invention, which can be obtained by those skilled in the art through current or future technologies.

[0022] In another aspect, the present invention provides the use of the above-mentioned Lactobacillus acidophilus LIHUO 1978 or the above-mentioned Lactobacillus acidophilus LIHUO 1978 preparation or the above-mentioned microbial agent in the preparation of a medicament for treating diarrhea.

[0023] Furthermore, the diarrhea described is chemotherapy-induced diarrhea.

[0024] Specifically, the viable count of Lactobacillus acidophilus LIHUO 1978 in the drug is 1.00 × 10⁻⁶. 6 -1.00×10 12CFU / mL or 1.00×10 6 -1.00×10 12 CFU / g.

[0025] Furthermore, the viable count of Lactobacillus acidophilus LIHUO 1978 in the drug is 5.00 × 10⁻⁶. 9 CFU / mL or 5.00×10 9 CFU / g.

[0026] Specifically, the drug has at least one of the following effects:

[0027] (1) Significantly reduced diarrhea scores;

[0028] (2) Significantly increases the length of the small intestine;

[0029] (3) Significantly reduced jejunal tissue pathology score;

[0030] (4) Significantly reduced serum interleukin-1β (IL-1β) concentration;

[0031] (5) Significantly increases serum IL-10 concentration.

[0032] In another aspect, the present invention provides a drug for relieving diarrhea, wherein the drug comprises the above-mentioned Lactobacillus acidophilus LIHUO 1978 or the above-mentioned Lactobacillus acidophilus LIHUO 1978 preparation or the above-mentioned microbial agent.

[0033] Specifically, the diarrhea described is chemotherapy-induced diarrhea.

[0034] Specifically, the drug may also include pharmaceutical excipients.

[0035] Furthermore, the pharmaceutical excipients are selected from any one or more of adjuvants, stabilizers, protectants, antibacterial agents, excipients, solubilizers, flavoring agents, diluents, and buffers.

[0036] Specifically, the dosage form of the drug is any one of tablets, capsules, granules, ophthalmic preparations, nasal preparations, suppositories, pills, ointments, creams, sprays, gels, powders, ointments, or lyophilized preparations.

[0037] In another aspect, the present invention provides the use of the above-mentioned Lactobacillus acidophilus LIHUO 1978 or the above-mentioned Lactobacillus acidophilus LIHUO 1978 preparation or the above-mentioned microbial agent in the preparation of products for improving constipation.

[0038] In another aspect, the present invention provides a product for improving constipation, wherein the product comprises the above-mentioned Lactobacillus acidophilus LIHUO 1978 or the above-mentioned Lactobacillus acidophilus LIHUO 1978 preparation or the above-mentioned microbial agent.

[0039] Specifically, the products mentioned include health foods, animal nutrition products, or pharmaceuticals.

[0040] Furthermore, the aforementioned health foods and animal nutrition products also include excipients.

[0041] Furthermore, the excipients include additives.

[0042] Furthermore, the additives are selected from any one or more of the following: acidity regulators, anti-caking agents, defoamers, antioxidants, bleaching agents, leavening agents, colorants, color protectants, emulsifiers, enzyme preparations, flavor enhancers, coating agents, moisture retainers, nutrient fortifiers, preservatives, stabilizers and coagulants, sweeteners, thickeners, food flavorings, and processing aids for the food industry.

[0043] Furthermore, the animal nutrition products include any one or more of animal feed and pet treats.

[0044] Specifically, when the product is a pharmaceutical product, the pharmaceutical product may also include pharmaceutical excipients;

[0045] Furthermore, the pharmaceutical excipients are selected from any one or more of adjuvants, stabilizers, protectants, antibacterial agents, excipients, solubilizers, flavoring agents, diluents, and buffers.

[0046] Specifically, the dosage form of the medicine is any one or more of the following: tablets, capsules, granules, ophthalmic preparations, nasal preparations, suppositories, pills, ointments, creams, sprays, gels, powders, ointments, or lyophilized preparations.

[0047] Specifically, the viable count of Lactobacillus acidophilus LIHUO 1978 in the product is 1.00 × 10⁻⁶. 6 -1.00×10 12 CFU / mL or 1.00×10 6 -1.00×10 12 CFU / g.

[0048] Furthermore, the viable count of Lactobacillus acidophilus LIHUO 1978 in the product is 1.00 × 10⁻⁶. 8 CFU / mL or 1.00×10 8 CFU / g.

[0049] Specifically, the product has at least one of the following functions:

[0050] (1) Significantly increases fecal particle count and fecal moisture content;

[0051] (2) Significantly improves gastrointestinal propulsion rate;

[0052] (3) Significantly reduced serum endothelin-1 (ET-1), serum vasoactive intestinal peptide (VIP), serum somatostatin (SS), and serum nitric oxide synthase (iNOS) concentrations;

[0053] (4) Significantly increased serum motilin (MTL), serum substance P (SP), serum acetylcholine (Ach), colon tissue 5-hydroxytryptamine (5-HT), and colon tissue IL-10 concentrations;

[0054] (5) Significantly increases the content of mucus-secreting goblet cells;

[0055] (6) Significantly increased the concentration of aquaporin 9 (AQP9) in colonic tissue;

[0056] (7) Regulates intestinal flora;

[0057] (8) Significantly increases the concentration of isobutyric acid, isovaleric acid, and valeric acid in colon contents.

[0058] In another aspect, the present invention provides the use of the above-mentioned Lactobacillus acidophilus LIHUO 1978 or the above-mentioned Lactobacillus acidophilus LIHUO 1978 preparation or the above-mentioned microbial agent in the preparation of products for regulating intestinal flora.

[0059] Specifically, the product can improve the β-diversity of the cecal contents microbiome.

[0060] Furthermore, the product can significantly increase the relative abundance of Lactobacillus and Lactobacillus mucosae in the gut.

[0061] In another aspect, the present invention provides a product for regulating intestinal flora, the product comprising the above-mentioned Lactobacillus acidophilus LIHUO 1978 or the above-mentioned Lactobacillus acidophilus LIHUO 1978 preparation or the above-mentioned microbial agent.

[0062] Specifically, the products mentioned include health foods, animal nutrition products, or pharmaceuticals.

[0063] Furthermore, the aforementioned health foods and animal nutrition products also include excipients that can be added to food.

[0064] Furthermore, the additives are selected from any one or more of the following: acidity regulators, anti-caking agents, defoamers, antioxidants, bleaching agents, leavening agents, colorants, color protectants, emulsifiers, enzyme preparations, flavor enhancers, coating agents, moisture retainers, nutrient fortifiers, preservatives, stabilizers and coagulants, sweeteners, thickeners, food flavorings, and processing aids for the food industry.

[0065] Furthermore, the health food includes probiotic products and nutritional supplements, and the animal nutrition products include any one or more of animal feed and pet snacks.

[0066] Specifically, when the product is a pharmaceutical product, the pharmaceutical product may also include pharmaceutical excipients;

[0067] Furthermore, the pharmaceutical excipients are selected from any one or more of adjuvants, stabilizers, protectants, antibacterial agents, excipients, solubilizers, flavoring agents, diluents, and buffers.

[0068] Furthermore, the dosage form of the medicine is any one or more of the following: tablets, capsules, granules, ophthalmic preparations, nasal preparations, suppositories, pills, ointments, creams, sprays, gels, powders, ointments, or lyophilized agents.

[0069] Furthermore, the viable count of Lactobacillus acidophilus LIHUO 1978 in the product is 1.00 × 10⁻⁶. 6 -1.00×10 12 CFU / mL or 1.00×10 6 -1.00×10 12 CFU / g.

[0070] Furthermore, the viable count of Lactobacillus acidophilus LIHUO 1978 in the product is 1.00 × 10⁻⁶. 8 CFU / mL or 1.00×10 8 CFU / g.

[0071] The beneficial effects of this invention are as follows:

[0072] (1) The Lactobacillus acidophilus LIHUO 1978 of the present invention has good safety, in vitro probiotic properties and good in vitro immunomodulatory properties.

[0073] (2) The effect of Lactobacillus acidophilus LIHUO 1978 in relieving chemotherapy-induced diarrhea is specifically manifested in the following ways: live Lactobacillus acidophilus LIHUO 1978 can significantly reduce the diarrhea score of chemotherapy-induced diarrhea mice, significantly increase the small intestine length of chemotherapy-induced diarrhea mice, significantly reduce the jejunal tissue pathology score of chemotherapy-induced diarrhea mice, significantly reduce the serum IL-1β concentration of chemotherapy-induced diarrhea mice, and significantly increase the serum IL-10 concentration of chemotherapy-induced diarrhea mice.

[0074] (3) The specific effects of Lactobacillus acidophilus LIHUO 1978 in improving constipation are as follows: both live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 significantly increase the number of fecal particles, fecal water content, and gastrointestinal propulsion rate in constipated rats; significantly regulate serum hormone levels in constipated rats: significantly reduce ET-1, VIP, and SS concentrations, and significantly increase MTL and SP concentrations; significantly increase neurotransmitter levels in constipated rats: significantly increase serum Ach concentration and colonic tissue 5-HT concentration; increase the content of secretory mucus goblet cells and colonic tissue AQP9 concentration in constipated rats; reduce serum iNOS concentration and significantly increase colonic tissue IL-10 concentration in constipated rats; and regulate intestinal flora. Live Lactobacillus acidophilus LIHUO 1978 can increase the concentrations of isobutyric acid, isovaleric acid, and valeric acid in constipated rats and improve short-chain fatty acid levels to relieve constipation.

[0075] Preservation Instructions

[0076] Accession number: GDMCC No: 65514;

[0077] Category Naming: Lactobacillus acidophilus LIHUO 1978;

[0078] Chinese name of the strain: Lactobacillus acidophilus LIHUO 1978;

[0079] Preservation period: November 20, 2024;

[0080] Preservation institution: Guangdong Provincial Center for Microbial Culture Collection;

[0081] Abbreviation of depositary institution: GDMCC;

[0082] Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Attached Figure Description

[0083] Figure 1 This is a phylogenetic tree constructed based on the 16S rDNA sequence of Lactobacillus acidophilus LIHUO 1978 in Example 1. The scale bar in the figure indicates 0.005, and the superscript "T" indicates the type strain.

[0084] Figure 2 The images show the colony morphology (left) and cell morphology (right) of Lactobacillus acidophilus LIHUO 1978 in Example 1, where the scale bar in the cell morphology image indicates 5 μm.

[0085] Figure 3The growth curves of *Lactobacillus acidophilus* LIHUO 1978 and *Lactobacillus acidophilus* NCFM in MRS liquid medium in Example 3 are shown above (OD). 600 Value curve, below: viable cell count curve).

[0086] Figure 4 The pH curves of Lactobacillus acidophilus LIHUO 1978 and Lactobacillus acidophilus NCFM cultured in MRS liquid medium in Example 3 are shown.

[0087] Figure 5 The acidity values ​​of Lactobacillus acidophilus LIHUO 1978 and Lactobacillus acidophilus NCFM fermented in skim milk in Example 3 are shown.

[0088] Figure 6 The concentrations of TNF-α and IL-10 produced by macrophages under normal conditions in each group of Example 5 are represented by ***. P <0.001.

[0089] Figure 7 The concentrations of TNF-α and IL-10 produced by macrophages in each group under inflammatory conditions in Example 5 are shown in **. P <0.01, *** indicates P <0.001.

[0090] Figure 8 The * indicates the TNF-α concentration in each group of the Caco-2 cell-macrophage co-culture system in Example 5. P <0.05.

[0091] Figure 9 The diarrhea scores of mice in each group in Example 6 are shown in **. P <0.01, *** indicates P <0.001.

[0092] Figure 10 The length of the small intestine in each group of mice in Example 6 is represented by ns. P >0.05, ** indicates P <0.01, *** indicates P <0.001.

[0093] Figure 11 The HE staining results of mouse jejunal tissues in each group in Example 7 are shown. The scale bar represents 200 μm.

[0094] Figure 12 The histopathological scores of the jejunum tissue of mice in each group in Example 7 are represented by ***. P <0.001.

[0095] Figure 13The concentrations of IL-1β and IL-10 in the serum of mice in each group in Example 8 are represented by ns. P >0.05, ** indicates P <0.01, *** indicates P <0.001.

[0096] Figure 14 The number of fecal particles in each group of rats in Example 9 is represented by ns. P >0.05, * indicates P <0.05, ** indicates P <0.01, *** indicates P <0.001.

[0097] Figure 15 The water content of feces in each group of rats in Example 9 is represented by ***. P <0.001.

[0098] Figure 16 ** represents the gastrointestinal propulsion rate of rats in each group in Example 9. P <0.01, *** indicates P <0.001.

[0099] Figure 17 The concentrations of ET-1, VIP, SS, MTL, and SP in the serum of rats in each group in Example 10 are represented by ns. P >0.05, * indicates P <0.05, ** indicates P <0.01, *** indicates P <0.001.

[0100] Figure 18 The values ​​represent the serum ACh concentration and colon tissue 5-HT concentration of rats in each group in Example 11. * indicates... P <0.05, ** indicates P <0.01, *** indicates P <0.001.

[0101] Figure 19 The results of AB staining of rat colon tissue in each group in Example 12 are shown. The scale bar represents 500 μm.

[0102] Figure 20 The concentration of AQP9 in the colon of rats in each group in Example 12 is indicated by **. P <0.01, *** indicates P <0.001.

[0103] Figure 21 The concentrations of iNOS in the serum of rats in each group in Example 13 are represented by ***. P<0.001.

[0104] Figure 22 The concentration of IL-10 in the colon of rats in each group in Example 13 is expressed in ns. P >0.05, * indicates P <0.05, *** indicates P <0.001.

[0105] Figure 23 The Ace index and Chao index of the cecal contents microbiome of each group of rats in Example 14 are represented by ns. P >0.05, * indicates P <0.05.

[0106] Figure 24 PCA and PCoA analyses of the cecal contents of rats in each group in Example 14 were performed at the genus level.

[0107] Figure 25 The relative abundance of microbiome genera in the cecal contents of rats in Example 14.

[0108] Figure 26 This study analyzed the differences in the genus level of the cecal contents of rats in different groups during Example 14.

[0109] Figure 27 The concentrations of isobutyric acid, isovaleric acid, and valerate in the colonic contents of rats in each group in Example 15 are represented by ns. P >0.05, * indicates P <0.05, ** indicates P <0.01. Detailed Implementation

[0110] To make the technical means, creative features, and achieved objectives and effects of this invention easier to understand, the invention is further illustrated below with specific embodiments. However, the following embodiments are merely preferred embodiments of this invention and not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments described herein without creative effort are all within the protection scope of this invention. Unless otherwise specified, the operating methods and equipment used in the following embodiments are conventional operating methods.

[0111] The strain of Lactobacillus acidophilus NCFM in the following examples has been disclosed in patent CN108102960A.

[0112] Example 1: Isolation, purification, identification, and preservation of Lactobacillus acidophilus LIHUO 1978

[0113] Experimental methods:

[0114] The Lactobacillus acidophilus LIHUO 1978 in this embodiment was isolated, purified, and identified in the following manner:

[0115] (1) Isolation and purification of Lactobacillus acidophilus LIHUO 1978: Samples of light cream collected from farms in Bayannur City, Inner Mongolia Autonomous Region, were rapidly transported back to the laboratory at low temperature for strain isolation experiments. The collected light cream samples were serially diluted with sterile physiological saline, and 10... -4 10 -5 Gradually diluted solutions were plated onto MRS solid medium and incubated anaerobically at 37°C for 48 hours. Based on morphological characteristics, dominant colonies were selected and placed in MRS liquid medium, incubated anaerobically at 37°C for 24 hours, and then streaked onto MRS solid medium and incubated anaerobically at 37°C for 48 hours. This streaking process was repeated 5 times until single colonies appeared on the medium.

[0116] (2) Identification of Lactobacillus acidophilus LIHUO 1978: 1.00 mL of the culture medium of the isolated strain was anaerobically cultured at 37℃ for 24 h. After centrifugation and washing with sterile PBS, genomic DNA of the strain was extracted using a DNA extraction kit (manufacturer: Beijing Tiangen Biotech, catalog number: Cat.#DP302-02). PCR amplification of the 16S rDNA sequence of the isolated strain was carried out using universal primers for bacterial 16S rDNA (27F, 1492R). The successfully amplified PCR product was sent to Shanghai Saiheng Biotechnology Co., Ltd. for sequencing. BLAST alignment using the NCBI GeneBank database was performed to obtain sequence homology, and the strain was identified as Lactobacillus acidophilus, named Lactobacillus acidophilus LIHUO 1978. A phylogenetic tree of the strain was constructed using MEGA software and the ortho-ligation method based on the 16S rDNA of Lactobacillus acidophilus LIHUO 1978.

[0117] (3) The colony morphology and cell morphology of Lactobacillus acidophilus LIHUO 1978, as well as the physiological and biochemical characteristics, were observed by CICC. The detection method referred to FMIC-QO01-001-2015 Microbiology Detection Bacterial Multiple Identification Detection Method.

[0118] (4) Preservation of Lactobacillus acidophilus LIHUO 1978: The identified Lactobacillus acidophilus LIHUO 1978 was deposited at the Guangdong Provincial Microbial Culture Collection Center (Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou).

[0119] Experimental results:

[0120] The sequencing results of the PCR product of 16S rDNA from *Lactobacillus acidophilus* LIHUO 1978 were compared with the NCBI GeneBank database using BLAST, showing that the similarity between this strain and the 16S rDNA sequence of *Lactobacillus acidophilus* was 99.73%. The sequence of the 16S rDNA of *Lactobacillus acidophilus* LIHUO 1978 is shown in SEQ ID NO.1:

[0121]

[0122] A phylogenetic tree constructed based on the 16S rDNA sequence of Lactobacillus acidophilus LIHUO 1978 is shown below. Figure 1 .

[0123] The colony morphology of Lactobacillus acidophilus LIHUO 1978 on MRS medium is as follows: white, round, moist, opaque, with regular edges (see...). Figure 2 (See the left image in the image). The bacterial morphology of this strain on MRS medium is that the cells are rod-shaped, arranged singly or in pairs (see image). Figure 2 (See the right image in the image) This strain is a Gram-positive strain.

[0124] The physiological and biochemical characteristics of Lactobacillus acidophilus LIHUO 1978 are shown in Table 1 ("+" indicates positive and "-" indicates negative).

[0125] Table 1. Physiological and biochemical characteristics of Lactobacillus acidophilus LIHUO 1978

[0126]

[0127] Example 2 Safety evaluation of Lactobacillus acidophilus LIHUO 1978

[0128] Experimental methods:

[0129] (1) Hemolytic test of Lactobacillus acidophilus LIHUO 1978: The hemolytic test of Lactobacillus acidophilus LIHUO 1978 was entrusted to CICC. The test method referred to FMIC-QO01-043-2024 Microbiological Detection Method for Hemolytic Detection of Microbial Strains.

[0130] (2) Detection of biogenic amines in Lactobacillus acidophilus LIHUO 1978: The detection of biogenic amines in Lactobacillus acidophilus LIHUO 1978 was entrusted to CICC. The detection method refers to GB 5009.208-2016 National Food Safety Standard Determination of Biogenic Amines in Food (Method I).

[0131] (3) Animal pathogenicity test of Lactobacillus acidophilus LIHUO 1978: The animal pathogenicity test of Lactobacillus acidophilus LIHUO 1978 was entrusted to CICC. The test method refers to Appendix A of the Technical Guidelines for Safety Inspection and Evaluation of Microbial Strains for Health Food Raw Materials (2020 Edition): Methods for Detection of Bacterial Pathogenicity in Health Food Raw Materials.

[0132] The activated strain of *Lactobacillus acidophilus* LIHUO 1978 was transferred to MRS agar plates containing 500 μg / mL cysteine ​​hydrochloride. Colonies on the agar plates were scraped and suspended in sterile physiological saline to prepare a concentration of 5.8 × 10⁻⁶. 7 A bacterial suspension at CFU / mL was administered via intraperitoneal injection to mice. Each mouse received a single injection of 0.2 mL of the suspension, and was observed for 21 consecutive days after the injection. The broth culture was prepared to a concentration of 3.3 × 10⁻⁶ CFU / mL. 8 The bacterial suspension had a concentration of 1.7 × 10⁻⁶ CFU / mL. 9 The 5-fold concentrated bacterial suspension at CFU / mL was administered to the corresponding groups of test mice by gavage at a dose of 20.0 mL / kg·BW once a day for 3 consecutive days. The mice were observed for 21 consecutive days starting from the first day of gavage.

[0133] Experimental results:

[0134] The hemolytic reaction of Lactobacillus acidophilus LIHUO 1978 was negative.

[0135] No putrescine, cadaverine, histamine, or tyramine were detected in Lactobacillus acidophilus LIHUO 1978, indicating that it does not produce biogenic amines.

[0136] Compared with the corresponding control groups, there were no significant differences in the initial and final body weights of the mice in each experimental group of Lactobacillus acidophilus LIHUO 1978. P＞ The concentration of *Lactobacillus acidophilus* LIHUO 1978 was 0.05%, and no adverse effects on the general health of the animals were observed, nor were any toxic reactions or deaths observed in the test mice. Therefore, strain *Lactobacillus acidophilus* LIHUO 1978 is non-pathogenic.

[0137] A good safety profile is a prerequisite for probiotics. The experimental results of this example show that Lactobacillus acidophilus LIHUO 1978 has a negative hemolytic reaction, and no putrescine, cadaverine, histamine, or tyramine were detected. It is also non-pathogenic, indicating that this strain has good safety and can be used for further development.

[0138] Example 3: Determination of growth characteristics, acid production characteristics, and skim milk fermentation characteristics of Lactobacillus acidophilus LIHUO 1978

[0139] Experimental methods:

[0140] (1) Activation of Lactobacillus acidophilus LIHUO 1978 and control strain Lactobacillus acidophilus NCFM

[0141] Lactobacillus acidophilus LIHUO 1978 and the control strain Lactobacillus acidophilus NCFM were inoculated into MRS liquid medium and cultured anaerobically at 37°C for 16 h (this was the first activation of the strain). The strains were activated three times in sequence.

[0142] (2) Determination of growth curves and pH curves of Lactobacillus acidophilus LIHUO 1978 and control strain Lactobacillus acidophilus NCFM: After three activations, Lactobacillus acidophilus LIHUO 1978 and control strain Lactobacillus acidophilus NCFM were inoculated into MRS liquid medium (pH 6.70) at a 3% inoculum and cultured anaerobically at 37℃ for 24 h. During this period, samples were taken every 2 h (3 replicates) to determine the OD of the strain culture medium. 600 The viable count of the bacterial culture medium was determined by plate counting method every 4 hours (3 replicates) to obtain the pH value and the viable count of the bacterial culture medium. The growth curves and pH curves of Lactobacillus acidophilus LIHUO 1978 and the control strain Lactobacillus acidophilus NCFM were plotted.

[0143] (3) Determination of the fermentation characteristics of skim milk by Lactobacillus acidophilus LIHUO 1978 and control strain Lactobacillus acidophilus NCFM: After activation three times, Lactobacillus acidophilus LIHUO 1978 and control strain Lactobacillus acidophilus NCFM were inoculated into 15% skim milk culture medium at an inoculation rate of 3% and cultured anaerobically at 37℃ for 24h. Samples were taken every 2h during the period (3 replicates). The total acid content of the samples was determined according to the national standard method for determination of food acidity (GB5009.239-2016).

[0144] Experimental results:

[0145] From 0h to 24h, with increasing culture time, the OD values ​​of *Lactobacillus acidophilus* LIHUO 1978 and the control strain *Lactobacillus acidophilus* NCFM in MRS liquid medium... 600 Both the value and the number of viable bacteria showed an upward trend, and the OD value... 600 The trend of changes in values ​​and viable cell counts is generally consistent (see...). Figure 3 When the culture time was 24 hours, the viable count of Lactobacillus acidophilus LIHUO 1978 was (2.12±0.08)×10⁻⁶. 9 The CFU / mL concentration was higher than that of the control strain Lactobacillus acidophilus NCFM (viable count was (1.98±0.03)×10⁻⁶). 9 CFU / mL.

[0146] From 0h to 24h, with increasing culture time, the pH value of both *Lactobacillus acidophilus* LIHUO 1978 and the control strain *Lactobacillus acidophilus* NCFM in MRS liquid medium showed a decreasing trend (see...). Figure 4After 12 hours of cultivation, the pH of the Lactobacillus acidophilus LIHUO 1978 culture decreased to 4.42±0.01, which was lower than that of the control strain Lactobacillus acidophilus NCFM (pH 4.57±0.00); after 24 hours of cultivation, the pH of the Lactobacillus acidophilus LIHUO 1978 culture decreased to 3.94±0.01, which was lower than that of the control strain Lactobacillus acidophilus NCFM (pH 4.00±0.02).

[0147] From 0h to 24h, the acidity values ​​of both *Lactobacillus acidophilus* LIHUO 1978 and the control strain *Lactobacillus acidophilus* NCFM showed an increasing trend during fermentation in skim milk; during the fermentation period (0h-24h), the acidity value of *Lactobacillus acidophilus* LIHUO 1978 was consistently higher than that of the control strain *Lactobacillus acidophilus* NCFM (see...). Figure 5 When cultured for 24 hours, the acidity of the skim milk system of Lactobacillus acidophilus LIHUO1978 was (151.63±0.93) °T, which was higher than that of the control strain Lactobacillus acidophilus NCFM (acidity value of (129.13±0.38) °T).

[0148] The experimental results of this embodiment show that Lactobacillus acidophilus LIHUO 1978 has good growth characteristics, acid production characteristics, and skim milk fermentation characteristics. Good growth characteristics and skim milk fermentation are conducive to its industrialization and saving its production energy consumption. Organic acids can inhibit the growth of pathogenic bacteria, and good acid production characteristics are conducive to its inhibition of intestinal pathogenic bacteria in the host, thereby reducing the probability of intestinal diseases such as diarrhea and constipation.

[0149] Example 4: Determination of tolerance of Lactobacillus acidophilus LIHUO 1978 and control strain Lactobacillus acidophilus NCFM in simulated gastrointestinal fluid.

[0150] Experimental methods:

[0151] (1) Preparation of test bacterial suspensions of Lactobacillus acidophilus LIHUO 1978 and control strain Lactobacillus acidophilus NCFM

[0152] The activation of Lactobacillus acidophilus LIHUO 1978 and the control strain Lactobacillus acidophilus NCFM was the same as in Example 3. After activation, the strains were washed three times with sterile physiological saline and the bacterial cells were resuspended in an appropriate amount of sterile physiological saline to prepare a bacterial suspension.

[0153] (2) Determination of tolerance of Lactobacillus acidophilus LIHUO 1978 and control strain Lactobacillus acidophilus NCFM in simulated gastrointestinal fluid.

[0154] 1.00 mL of Lactobacillus acidophilus LIHUO 1978 and the control strain Lactobacillus acidophilus NCFM were inoculated into 9.00 mL of simulated gastric fluid (pH 2.50, containing 1.00% pepsin) and anaerobic at 37°C for 3 h. Samples were taken at 0 h and 3 h, and the viable bacterial count (N0 and N1, respectively) was determined by plate counting method and the survival rate was calculated (the survival rate in simulated gastric fluid was N1 / N0×100%).

[0155] 1.00 mL of Lactobacillus acidophilus LIHUO 1978 and control strain Lactobacillus acidophilus NCFM, respectively, after being treated with simulated gastric fluid for 3 h, were transferred to 9.00 mL of simulated intestinal fluid (pH 8.00, containing 1.00% trypsin and 1.80% bile salts). After treatment at 37℃ for 8 h, samples were taken, and the viable count N2 was determined by plate counting method, and the survival rate was calculated (the survival rate in simulated intestinal fluid is N2 / N1×100%, and the survival rate in simulated gastrointestinal fluid is N2 / N0×100%).

[0156] Experimental results:

[0157] The survival rates of Lactobacillus acidophilus LIHUO 1978 and the control strain Lactobacillus acidophilus NCFM after treatment in simulated gastric fluid for 3 h, simulated intestinal fluid for 8 h, and simulated gastrointestinal fluid for 11 h were 95.18%, 84.71%, and 80.63%, respectively, all higher than the survival rate of the control strain Lactobacillus acidophilus NCFM (84.94%, 82.53%, and 70.10%, see Table 2).

[0158] The experimental results of this embodiment show that Lactobacillus acidophilus LIHUO 1978 has good tolerance in simulated gastrointestinal fluid, which is beneficial for the strain to resist the intestinal environment of low pH gastric acid and high concentration of bile salts, and is conducive to its probiotic effect in the host intestine.

[0159] Table 2. Survival rates of *Lactobacillus acidophilus* LIHUO 1978 and *Lactobacillus acidophilus* NCFM in simulated gastrointestinal fluid.

[0160]

[0161] Example 5 Evaluation of the in vitro immunomodulatory properties of Lactobacillus acidophilus LIHUO 1978

[0162] Experimental methods:

[0163] (1) Preparation of test bacterial suspension and heat-inactivated cells of Lactobacillus acidophilus LIHUO 1978

[0164] The activation of Lactobacillus acidophilus LIHUO 1978 was the same as in Example 3. After activation three times, the strain was washed three times with sterile PBS solution, and the cells were resuspended in an appropriate amount of sterile PBS solution to prepare a viable count of 1.00 × 10⁻⁶ cells / year. 7 CFU / mL of test bacterial solution; Lactobacillus acidophilus LIHUO 1978 test bacterial solution was sterilized at 121℃ for 15 min to prepare heat-inactivated bacterial cells.

[0165] (2) Effects of live Lactobacillus acidophilus LIHUO 1978, heat-inactivated bacteria, and macrophages in normal condition (the experiment was conducted according to the method described in patent CN119570698A): Macrophages were activated, and 1.00 × 10 5 Cells were inoculated into 48-well plates at a rate of 1 cell / well and incubated overnight at 37°C with 5% CO2. After cell adhesion, the supernatant was discarded, and 900 μL of DMEM culture medium was added to each well. For the control group, 100 μL of sterile PBS solution was added to each well; for the LIHUO 1978 group, 100 μL of *Lactobacillus acidophilus* LIHUO 1978 test bacterial culture was added to each well; for the heat-inactivated LIHUO1978 group, 100 μL of heat-inactivated *Lactobacillus acidophilus* LIHUO1978 bacterial cells were added to each well. Each group was repeated three times. After incubation at 37°C with 5% CO2 for 2 hours, the cell culture supernatant was collected from each well. The concentrations of TNF-α and IL-10 in the cell culture supernatant of each group were determined using an ELISA kit (manufacturer: Shanghai ELISA, catalog number: ml002095A) and an ELISA kit (manufacturer: Shanghai ELISA, catalog number: ml037873A).

[0166] (3) Effect of heat-inactivated Lactobacillus acidophilus LIHUO 1978 on macrophages in an inflammatory state (the experiment was conducted according to the method described in patent CN119570698A): Macrophages were activated and then subjected to 1.00×10 5Inoculate cells at a rate of 10 cells / well into 48-well plates and incubate overnight at 37°C with 5% CO2. After cell adhesion, discard the supernatant and add 900 μL of LMEM culture medium to each well. For the control group, add 100 μL of sterile PBS solution to each well; for the model group, add 100 μL of sterile PBS solution to each well; for the heat-inactivated LIHUO1978 group, add 100 μL of heat-inactivated Lactobacillus acidophilus LIHUO 1978 cells to each well. Each group is repeated three times. Incubate at 37°C with 5% CO2 for 2 hours. After incubation, aspirate the supernatant, wash three times with sterile PBS solution, and add 900 μL of LMEM culture medium to each well. For the control group, add 100 μL of sterile PBS solution to each well; for the model group, add 100 μL of LPS solution (10 μg / mL) to each well; for the heat-inactivated LIHUO1978 group, add 100 μL of LPS solution (10 μg / mL) to each well. The cells were incubated at 37℃ and 5% CO2 for 2 hours. The cell culture supernatant was collected from each well, and the concentrations of TNF-α and IL-10 in the cell culture supernatant of each group were determined using an enzyme-linked immunosorbent assay kit (ELISA kit) for TNF-α (manufacturer: Shanghai ELISA, catalog number: ml002095A) and IL-10 (manufacturer: Shanghai ELISA, catalog number: ml037873A).

[0167] (4) Effect of heat-inactivated Lactobacillus acidophilus LIHUO 1978 cells on the co-culture system of Caco-2 cells and macrophages (the experiment was conducted according to the method described in patent CN119570698A): Caco-2 cells were activated, and 1.00×10 5 Inoculate 1.00 × 10⁶ cells / well into a 24-well plate, and inoculate the Transwell chambers top-side down. 5 Caco-2 cells were inserted into 24-well plates with the reverse side of the chambers to establish a co-culture system. 500 μL of DMEM medium was added, and the plates were incubated at 37°C with 5% CO2. Once a dense monolayer of Caco-2 cells formed in the chambers, 200 μL of DMEM medium was added to each well. For the control group, 100 μL of sterile PBS solution was added to each well. For the heat-inactivated LIHUO1978 group, 100 μL of heat-inactivated Lactobacillus acidophilus LIHUO 1978 cells were added to each well, with three replicates per group. The plates were incubated at 37°C with 5% CO2 for 2 hours. After incubation, the lower chamber and cell culture supernatant were aspirated, and the concentration of TNF-α in the cell culture supernatant of each group was determined using a TNF-α (manufacturer: Shanghai Enzyme Linked Immunosorbent Assay, catalog number: ml002095A) ELISA kit.

[0168] Experimental results:

[0169] Compared with the control group, the concentrations of TNF-α and IL-10 produced by macrophages under normal conditions were significantly increased in the LIHUO 1978 group and the heat-inactivated LIHUO1978 group (see...). Figure 6 ).

[0170] Compared with the control group, the concentrations of TNF-α and IL-10 produced by macrophages under inflammatory conditions were significantly increased in the model group; compared with the model group, the concentrations of TNF-α and IL-10 produced by macrophages under inflammatory conditions were significantly increased in the heat-inactivated LIHUO1978 group (see...). Figure 7 ).

[0171] Compared with the control group, the TNF-α concentration in the Caco-2 cell-macrophage co-culture system of the heat-inactivated LIHUO1978 group was significantly increased (see...). Figure 8 ).

[0172] The experimental results of this embodiment show that live Lactobacillus acidophilus LIHUO 1978 can significantly promote the production of TNF-α and IL-10 by normal macrophages; heat-inactivated Lactobacillus acidophilus LIHUO 1978 can significantly promote the production of TNF-α and IL-10 by normal and inflammatory macrophages, and can significantly promote the production of TNF-α in the Caco-2 cell-macrophage co-culture system. Both live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 have good in vitro immunomodulatory properties, which is beneficial for them to exert immunomodulatory effects in the host intestine, thereby exerting their probiotic properties.

[0173] Example 6: Effects of live Lactobacillus acidophilus LIHUO 1978 on symptoms of chemotherapy-induced diarrhea in mice.

[0174] Experimental methods:

[0175] (1) Preparation of Lactobacillus acidophilus LIHUO 1978 bacterial culture

[0176] The activation of Lactobacillus acidophilus LIHUO 1978 was the same as in Example 3. After activation three times, the strain was washed three times with sterile PBS solution, and the cells were resuspended in an appropriate amount of sterile PBS solution to prepare a viable count of 5.00 × 10⁻⁶ cells. 9 CFU / mL bacterial culture

[0177] (2) Animal experiment design

[0178] SPF-grade 6-8 week old male BALB / c mice (weighing 18g-20g) provided by Changsha Tianqin Biotechnology Co., Ltd. were used as research subjects. All mice were placed in a stable environment with a temperature of (23±2)℃ and humidity of (50±5)%. Throughout the experiment, the mice were housed under 12-hour light-dark cycles, with free access to SPF-grade feed and sterile water. After 7 days of acclimatization, all mice were grouped using a weight-based serpentine grouping method (n=10 per group): blank group, model group, drug group, and LIHUO 1978 experimental group. Animal experiments were conducted according to Table 3.

[0179] Table 3. Animal experimental design for the use of Lactobacillus acidophilus LIHUO 1978 to alleviate chemotherapy-induced diarrhea.

[0180]

[0181] (3) Diarrhea score in mice

[0182] After the end of gavage and intraperitoneal injection on day 16, each mouse was placed in a clean container with a clean filter paper at the bottom. The mice were allowed to defecate, and the fecal condition was observed. The mice were then scored for diarrhea according to the scoring rules in Table 4.

[0183] Table 4 Diarrhea Scoring Table

[0184]

[0185] (4) Anatomy of mice and determination of small intestine length

[0186] All mice were fasted and deprived of water for 12 hours after gavage, intraperitoneal injection, and diarrhea scoring on day 16. They were then anesthetized with isoflurane and euthanized. Blood samples were collected from the eyes of all mice, and the abdomens were disinfected with 75% ethanol solution. Dissection experiments were then performed, and the entire small intestine was removed and its total length was measured with a ruler.

[0187] After washing the jejunal tissue of each group of mice with cold physiological saline, about 0.5 cm of distal jejunal tissue was cut and fixed in 4% paraformaldehyde solution.

[0188] After the blood samples from each group of mice were allowed to stand and separate into layers, serum was obtained by centrifugation at 4℃ and 3000r / min for 15min, aliquoted, and stored at -80℃ for testing.

[0189] Experimental results:

[0190] Compared with the control group, the diarrhea scores of mice in the model group were significantly increased; compared with the model group, the diarrhea scores of mice in the drug group and the LIHUO1978 experimental group were significantly decreased by 29.17% and 37.50%, respectively (see...). Figure 9 ).

[0191] Compared with the control group, the small intestine length of mice in the model group was significantly shortened by 16.80%; compared with the model group, the small intestine length of mice in the drug group was not significantly different, while the small intestine length of mice in the LIHUO 1978 experimental group was significantly increased by 9.29% (see...). Figure 10 ).

[0192] The experimental results of this embodiment show that Lactobacillus acidophilus LIHUO 1978 can significantly reduce diarrhea scores and significantly increase small intestinal length, thereby alleviating the symptoms of chemotherapy-induced diarrhea in mice.

[0193] Example 7: Effects of Lactobacillus acidophilus LIHUO 1978 on jejunal histological damage in mice with chemotherapy-induced diarrhea.

[0194] Experimental methods:

[0195] In Example 6, all mouse jejunal tissues fixed in 4% paraformaldehyde solution were dehydrated, embedded in paraffin, sectioned at 5μm, dewaxed, and stained with hematoxylin and eosin (H&E staining). The damage to the mouse jejunal tissues was observed under a microscope, and the pathological scores of the jejunal tissues were calculated according to the following criteria: (1) mononuclear cell infiltration or mixed infiltration of neutrophils in the mucosa; (2) thinning of the mucosa or glands, reduction of goblet cells, and crypts and glands being far from the basement membrane; (3) disappearance of crypts, with neutrophils being dominant; (4) epithelial destruction, increased basal plasma cells, and lymphocyte aggregation. The pathological scoring criteria were that each of the above pathological manifestations was scored as 1 point, and if more than 3 of the above pathological manifestations were present, the total score was 4 points.

[0196] The experimental results are as follows:

[0197] HE staining results of jejunal tissues from each group of mice are shown in the figure. Figure 11 The results showed that the jejunal tissue of mice in the blank control group was rich in intestinal villi, which were large, well-arranged, and regularly arranged. Goblet cells were also abundant, with no obvious histopathological changes. The jejunal tissue of mice in the model group exhibited severe structural abnormalities, with significant atrophy and disappearance of intestinal villi, epithelial cell edema, a significant reduction in the number of goblet cells, extensive destruction and disappearance of numerous intestinal crypt structures, disordered arrangement, and extensive inflammatory cell infiltration. The jejunal tissue of mice in the drug group showed mild structural abnormalities, with slight atrophy and slightly disordered arrangement of intestinal villi, a reduced number of goblet cells, disordered crypt arrangement, moderate mucosal edema, and moderate inflammatory cell infiltration. The LIHUO 1978 experimental group of mice showed mild structural abnormalities in their jejunal tissue, with slight atrophy and slightly disordered arrangement of intestinal villi, abundant goblet cells, and mild inflammatory cell infiltration in the mucosal layer.

[0198] Compared with the control group, the histopathological score of the jejunum tissue of mice in the model group was significantly increased; compared with the model group, the histopathological scores of the intestinal tissue of mice in the drug group and the LIHUO 1978 experimental group were significantly decreased by 30.77% and 53.85%, respectively (see...). Figure 12 ).

[0199] The experimental results of this embodiment show that Lactobacillus acidophilus LIHUO 1978 can significantly improve the degree of jejunal tissue lesions in mice with chemotherapy-induced diarrhea, thereby alleviating chemotherapy-induced diarrhea.

[0200] Example 8: Effects of Lactobacillus acidophilus LIHUO 1978 on serum inflammatory factors in mice with chemotherapy-induced diarrhea.

[0201] Experimental methods:

[0202] The concentrations of IL-1β and IL-10 in the serum of mice in each group in Example 6 were determined using an IL-1β (manufacturer: Shanghai Enzyme Linked, catalog number: ml098416A) and IL-10 (manufacturer: Shanghai Enzyme Linked, catalog number: ml037873A) enzyme-linked immunosorbent assay kit and strictly in accordance with the instructions.

[0203] The experimental results are as follows:

[0204] Compared with the control group, the serum IL-1β concentration in the model group mice increased significantly by 98.41%; compared with the model group, the serum IL-1β concentration in the drug group and the LIHUO 1978 experimental group mice decreased significantly by 27.39% and 35.19%, respectively (see...). Figure 13 ).

[0205] Compared with the blank group, the serum IL-10 concentration in the model group mice decreased significantly by 27.69%; compared with the model group, the serum IL-10 concentration in the drug group mice increased by 19.70%, but the difference was not significant, while the serum IL-10 concentration in the LIHUO 1978 experimental group mice increased significantly by 55.82%. Figure 13 ).

[0206] The experimental results of this embodiment show that Lactobacillus acidophilus LIHUO 1978 can significantly reduce the concentration of the pro-inflammatory factor IL-1β and significantly increase the concentration of the anti-inflammatory factor IL-10 in the serum of mice with chemotherapy-induced diarrhea, thereby regulating the concentration of serum inflammatory factors and alleviating chemotherapy-induced diarrhea.

[0207] Example 9: Effects of live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria on constipation symptoms in rats.

[0208] Experimental methods:

[0209] (1) Preparation of Lactobacillus acidophilus LIHUO 1978 bacterial suspension and heat-inactivated bacterial cells

[0210] The activation of Lactobacillus acidophilus LIHUO 1978 was the same as in Example 3. After activation three times, the strain was washed three times with sterile PBS solution, and the cells were resuspended in an appropriate amount of sterile PBS solution to prepare a viable count of 1.00 × 10⁻⁶ cells. 8 CFU / mL bacterial suspension; Lactobacillus acidophilus LIHUO 1978 bacterial suspension was sterilized at 121℃ for 15 min to prepare heat-inactivated bacterial cells.

[0211] (2) Animal experiment design

[0212] Six- to eight-week-old SPF-grade male SD rats (weighing 180-220g) provided by Changsha Tianqin Biotechnology Co., Ltd. were used as the research subjects. All rats were placed in a stable environment with a temperature of (23±2)℃ and humidity of (50±5)%. Throughout the experiment, the rats were housed under 12-hour light-dark cycles, with free access to SPF-grade feed and sterile water. After 7 days of acclimatization, all rats were grouped using a weight-based serpentine grouping method (n=8 per group): normal group, constipation model group, polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group. Animal experiments were conducted according to Table 5.

[0213] Table 5. Animal experimental design for relieving constipation with live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria.

[0214]

[0215] (3) Determination of rat fecal particle count and water content

[0216] On day 21, feces excreted by each rat in all groups within 1 hour were collected, the number of fecal particles was recorded, the feces were weighed (m1) and dried in an oven at 60℃ to constant weight (m2), and the fecal moisture content of each rat was calculated according to the formula: fecal moisture content (%) = (m1-m2) / m1×100%.

[0217] (4) Rat anatomy and determination of rat gastrointestinal propulsion rate

[0218] All rats were fasted and deprived of water for 12 hours after gavage and intraperitoneal injection on day 21. Each rat was gavaged with 2 mL of Evans blue paste (the preparation method of Evans blue paste was carried out according to the method in patent CN119331785B: Evans blue paste was prepared by aseptically mixing 2.5 g carboxymethyl cellulose, 8 g milk powder, 4 g sucrose, 4 g starch, and 3.5 mL of 1% Evans blue solution to form 150 mL of Evans blue semi-solid paste). Blood samples were collected by puncturing the eye sockets at 15 minutes, and the rats were then sacrificed. A necropsy experiment was performed on the rats, and the complete gastrointestinal tissue was removed. The distance from the pyloric sphincter to the end of the Evans blue paste and the total length of the small intestine were measured and recorded using a ruler. The gastrointestinal propulsion rate (%) was calculated for each rat using the formula: Gastrointestinal propulsion rate (%) = Distance of Evans blue paste to the end of the paste (cm) / Total length of small intestine (cm) × 100%.

[0219] After washing the colon tissues of rats in each group with cold saline, about 1 cm of distal colon tissue was cut off and fixed in 4% paraformaldehyde solution; the remaining colon tissues were collected into sterile enzyme-free cryovials, flash-frozen in liquid nitrogen, and stored at -80℃ for testing.

[0220] After the blood samples from each group of rats were allowed to stand and separate into layers, serum was obtained by centrifugation at 3000 r / min for 15 min at 4℃, aliquoted, and stored at -80℃ for testing.

[0221] Under aseptic conditions, the contents of the cecum and colon of rats in each group were collected, flash-frozen in liquid nitrogen, and stored at -80℃ for analysis.

[0222] The experimental results are as follows:

[0223] Compared with the normal group, the number of fecal particles in the constipation model group rats decreased significantly by 53.85%; compared with the constipation model group, the number of fecal particles in the polyethylene glycol 4000 powder group rats increased by 79.33%, but the difference was not significant; while the number of fecal particles in the LIHUO 1978 group and the heat-inactivated LIHUO 1978 group rats increased significantly by 137.67% and 100.00%, respectively (see...). Figure 14 ).

[0224] Compared with the normal group, the fecal water content of rats in the constipation model group decreased significantly by 29.08%; while compared with the constipation model group, the fecal water content of rats in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group increased significantly by 32.75%, 34.27%, and 33.18%, respectively (see...). Figure 15 ).

[0225] Compared with the normal group, the gastrointestinal propulsion rate of rats in the constipation model group decreased significantly by 42.44%; while compared with the constipation model group, the gastrointestinal propulsion rate of rats in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group increased significantly by 22.73%, 65.19%, and 61.48%, respectively (see...). Figure 16 ).

[0226] The experimental results of this embodiment show that the number of fecal particles, fecal water content, and gastrointestinal propulsion rate of rats in the constipation model group were significantly lower than those in the normal group, indicating that the constipation rat model was successfully established. Both live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria can alleviate constipation symptoms in rats by significantly increasing the number of fecal particles, fecal water content, and gastrointestinal propulsion rate.

[0227] Example 10 Effects of live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria on serum ET-1, VIP, SS, MTL, and SP concentrations in constipated rats

[0228] Experimental methods:

[0229] The concentrations of ET-1, VIP, SS, MTL, and SP in the serum of rats in Example 9 were determined using ET-1 (manufacturer: Shanghai Enzyme Linked, catalog number: ml002890V), VIP (manufacturer: Shanghai Enzyme Linked, catalog number: ml936589V), SS (manufacturer: Shanghai Enzyme Linked, catalog number: ml003102V), MTL (manufacturer: Shanghai Enzyme Linked, catalog number: ml003079V), and SP (manufacturer: Shanghai Enzyme Linked, catalog number: ml003119V) enzyme-linked immunosorbent assay kits, strictly following the instructions.

[0230] Experimental results:

[0231] Compared with the normal group, the serum ET-1 concentration in the constipation model group rats increased significantly by 56.82%; while compared with the constipation model group, the serum ET-1 concentration in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group rats decreased significantly by 12.35%, 16.60%, and 16.21%, respectively (see...). Figure 17 ).

[0232] Compared with the normal group, the serum VIP concentration in the constipation model group rats increased significantly by 150.29%; while compared with the constipation model group, the serum VIP concentration in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group rats decreased significantly by 20.32%, 31.18%, and 26.55%, respectively (see...). Figure 17 ).

[0233] Compared with the normal group, the serum SS concentration in the constipation model group of rats increased significantly by 133.43%; while compared with the constipation model group, the serum SS concentration in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group of rats decreased significantly by 27.28%, 33.78%, and 38.87%, respectively (see...). Figure 17 ).

[0234] Compared with the normal group, the serum MTL concentration of rats in the constipation model group decreased significantly by 40.46%; while compared with the constipation model group, the serum MTL concentration of rats in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group increased significantly by 27.97%, 39.48%, and 28.92%, respectively (see...). Figure 17 ).

[0235] Compared with the normal group, the SP concentration in the constipation model group rats decreased significantly by 44.00%; compared with the constipation model group, the SP concentration in the serum of the polyethylene glycol 4000 powder group rats increased by 16.15%, but the difference was not significant, while the SP concentration in the serum of the LIHUO 1978 group and the heat-inactivated LIHUO 1978 group rats increased significantly by 56.32% and 48.95%, respectively (see...). Figure 17 ).

[0236] ET-1 inhibits intestinal peristalsis by suppressing smooth muscle contraction, VIP by dilating blood vessels, and SS by inhibiting the secretion of gastrointestinal juices; MTL promotes intestinal peristalsis by enhancing gastrointestinal motility and emptying, and SP by stimulating intestinal contraction through activating intestinal smooth muscle. The experimental results of this embodiment show that both live and heat-inactivated *Lactobacillus acidophilus* LIHUO 1978 bacteria can significantly reduce serum ET-1, VIP, and SS concentrations in constipated rats, significantly increase serum MTL and SP concentrations in constipated rats, and regulate serum hormone levels in constipated rats, thereby alleviating constipation.

[0237] Example 11 Effects of live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria on serum acetylcholine concentration and colonic tissue 5-hydroxytryptamine concentration in constipated rats

[0238] Experimental methods:

[0239] (1) Determination of serum Ach concentration in rats

[0240] The concentration of Ach in the serum of rats in each group in Example 9 was determined using an Ach (manufacturer: Shanghai Enzyme Link, catalog number: ml003048V) enzyme-linked immunosorbent assay kit and strictly in accordance with the instructions.

[0241] (2) Determination of protein concentration in rat colon tissue

[0242] The colon tissues of rats in each group in Example 9 were homogenized and centrifuged to obtain supernatant. The protein concentration was measured using the BCA Protein Concentration Assay Kit (Enhanced Version) (manufacturer: Beyotime, catalog number: Cat No. P0010) and strictly in accordance with the instructions.

[0243] (3) Determination of 5-HT concentration in rat colon tissue

[0244] The concentration of 5-HT in the colon tissue of rats in each treatment group in Example 9 was determined using a 5-HT enzyme-linked immunosorbent assay kit (manufacturer: Shanghai Enzyme Link, catalog number: ml059511V) and strictly in accordance with the instructions.

[0245] Experimental results:

[0246] Compared with the normal group, the serum Ach concentration in the constipation model group was significantly decreased by 44.60%; while compared with the constipation model group, the serum Ach concentration in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group was significantly increased by 36.80%, 55.53%, and 47.07%, respectively (see...). Figure 18 ).

[0247] Compared with the normal group, the 5-HT concentration in the colonic tissue of rats in the constipation model group decreased significantly by 21.05%; while compared with the constipation model group, the 5-HT concentration in the colonic tissue of rats in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group increased significantly by 10.00%, 13.33%, and 10.00%, respectively (see...). Figure 18 ).

[0248] Ach can promote the secretion of gastric and intestinal juices, thereby promoting intestinal peristalsis. 5-HT is an important neurotransmitter that reduces the transit time of feces in the colon. The experimental results of this embodiment show that both live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria can significantly increase the serum Ach concentration and colonic tissue 5-HT concentration in constipated rats, thereby regulating the neurotransmitter levels in constipated rats and alleviating constipation.

[0249] Example 12 Effects of live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria on the content of mucus goblet cells and the level of aquaporin 9 in colonic tissue of constipated rats

[0250] Experimental methods:

[0251] (1) AB staining of rat colon tissue

[0252] All rat colon tissues fixed in 4% paraformaldehyde solution in Example 9 were sequentially dehydrated, paraffin-embedded, sectioned at 5 μm, dewaxed, stained with Alcian Blue (AB), and scanned for imaging to observe the distribution and content of secretory mucus goblet cells in the rat colon tissue.

[0253] (2) Determination of AQP9 concentration in rat colon tissue

[0254] The colon tissues of rats in each group in Example 9 were homogenized and centrifuged to obtain supernatant. The protein concentration of the colon tissues of rats in each group was measured in the same way as in Example 11. AQP9 (manufacturer: Shanghai Enzyme Linker, catalog number: ml059561V) was used and the AQP9 concentration of the colon tissues of rats in each treatment group in Example 9 was measured strictly in accordance with the instructions.

[0255] Experimental results:

[0256] The distribution and content of mucus-secreting goblet cells in the colon tissue of rats in each group were detected by AB staining (see AB staining). Figure 19 The results showed that, compared with the normal group, the content of secretory mucus goblet cells in the colonic tissue of rats in the constipation model group decreased; while compared with the constipation model group, the content of secretory mucus goblet cells in the colonic tissue of rats in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group all increased.

[0257] Compared with the normal group, the AQP9 concentration in the colonic tissue of rats in the constipation model group decreased significantly by 49.84%; while compared with the constipation model group, the AQP9 concentration in the colonic tissue of rats in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group increased significantly by 26.56%, 53.36%, and 60.24%, respectively (see...). Figure 20 ).

[0258] Mucus goblet cells in the colon secrete mucus proteins that protect the colonic mucosa and promote defecation. AQP9 in the colon can also promote defecation by participating in the secretion of colonic mucus proteins. The experimental results of this embodiment show that both live and heat-inactivated *Lactobacillus acidophilus* LIHUO 1978 bacteria can significantly increase the concentration of AQP9 in the colonic tissue of constipated rats by increasing the content of mucus-secreting goblet cells, thereby alleviating constipation.

[0259] Example 13 Effects of live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria on serum nitric oxide synthase concentration and colonic tissue IL-10 concentration in constipated rats

[0260] Experimental methods:

[0261] (1) Determination of serum iNOS concentration in rats

[0262] The concentration of iNOS in the serum of rats in each group in Example 9 was determined using an iNOS (manufacturer: Shanghai Enzyme Linker, catalog number: ml059045V) enzyme-linked immunosorbent assay kit and strictly in accordance with the instructions.

[0263] (2) Determination of IL-10 concentration in rat colon tissue

[0264] The rat colon tissues of each group in Example 9 were homogenized and centrifuged to obtain supernatant. The protein concentration of the rat colon tissues of each group was determined in the same way as in Example 11. The IL-10 concentration of the rat colon tissues of each treatment group in Example 9 was determined by using an IL-10 (manufacturer: Shanghai Enzyme Link, catalog number: ml002813V) enzyme-linked immunosorbent assay kit and strictly following the instructions.

[0265] The experimental results are as follows:

[0266] Compared with the normal group, the serum iNOS concentration in the constipation model group rats increased significantly by 521.21%; while compared with the constipation model group, the serum iNOS concentration in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group rats decreased significantly by 50.49%, 59.76%, and 54.63%, respectively (see...). Figure 21 ).

[0267] Compared with the normal group, the IL-10 concentration in the colonic tissue of rats in the constipation model group decreased significantly by 22.07%; compared with the constipation model group, the IL-10 concentration in the colonic tissue of rats in the polyethylene glycol 4000 powder group increased by 4.42%, but the difference was not significant; while the IL-10 concentration in the colonic tissue of rats in the LIHUO 1978 group and the heat-inactivated LIHUO 1978 group increased significantly by 29.20% and 17.70%, respectively (see...). Figure 22 ).

[0268] iNOS is associated with intestinal inflammation and can exacerbate constipation symptoms by producing excessive NO, leading to relaxation of intestinal smooth muscle. IL-10 is an anti-inflammatory factor that can alleviate intestinal inflammation caused by constipation. The experimental results of this embodiment show that both live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria can significantly reduce serum iNOS concentration and significantly increase colonic tissue IL-10 concentration in constipated rats, thereby regulating the levels of inflammation-related factors and relieving constipation.

[0269] Example 14 Effects of live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria on intestinal flora in constipated rats

[0270] Experimental methods:

[0271] In Example 9, cecal contents from six rats were randomly selected from each group, and Shanghai Meiji Biomedical Technology Co., Ltd. was commissioned to conduct microbial diversity detection. (The experimental method was carried out according to the method in patent CN119331785B.) Total DNA was extracted from the rat cecal contents, and the V3-V4 region of 16S rDNA was amplified using 338F and 806R. Based on different similarity levels, all sequences were divided into OTUs, and bioinformatics statistical analysis was performed on the OTUs at a 97% similarity level. Alpha diversity, β diversity, the proportion of different species in different groups, and species differences were analyzed, and plotted using R language.

[0272] The experimental results are as follows:

[0273] Compared with the normal group, there was no significant difference in the genus-level Ace index of the cecal contents microbiome of rats in the constipation model group; compared with the constipation model group, there were no significant differences in the genus-level Ace index of the cecal contents microbiome of rats in the polyethylene glycol 4000 powder group, LIHUO 1978 group, and heat-inactivated LIHUO 1978 group (see...). Figure 23 ).

[0274] Compared with the normal group, there was no significant difference in the Chao index of the cecal contents microbiome at the genus level in the constipation model group; compared with the constipation model group, the Chao index of the cecal contents microbiome at the genus level in the polyethylene glycol 4000 powder group was significantly increased, while there was no significant difference in the Chao index of the cecal contents microbiome at the genus level between the LIHUO 1978 group and the heat-inactivated LIHUO 1978 group (see...). Figure 23 ).

[0275] Principal component analysis (PCA) and principal coordinate analysis (PCoA) of the cecal contents microbiome at the genus level in each group of rats (see...) Figure 24 The results show that the normal group and the constipation model group are far apart, indicating that the composition of the microbiome of the cecal contents of the rats in the normal group and the constipation model group is different. The LIHUO 1978 group and the heat-inactivated LIHUO 1978 group are closer to the normal group and farther apart from the constipation model group, indicating that the composition of the microbiome of the cecal contents of the LIHUO 1978 group and the heat-inactivated LIHUO 1978 group is more similar to that of the normal group. Clostridia _UCG-014, Rombutzella genus ( Romboutsia ), Lactobacillus ( Lactobacillus The relative abundance of the microbiome in the cecal contents of rats in all groups was high, indicating that it was the dominant bacterial species (see...). Figure 25 The relative abundance of microorganisms at the cecal contents of rats in the LIHUO 1978 group and the heat-inactivated LIHUO 1978 group was closer to that in the normal group.

[0276] Compared with the normal group, the genus *Romebuts* in the cecal contents microbiome of the constipation model rats was significantly higher. Romboutsia Staphylococcus spp. Staphylococcus ), Mammalian cocci ( Mammaliicoccus ), Sea cocci ( Jeotgalicoccus Host-associated Lactobacillus genus ( Ligilactobacillus ), Zurich bacillus ( Turicibacter The relative abundance of norank_f_ decreased significantly. Muribaculaceae , Lachnospiraceae _NK4A136_group, Bacteroides genus ( Bacteroides ), [ Eubacterium ]_ siraeum The relative abundance of *Lactobacillus* spp. in the cecal contents of rats in the polyethylene glycol 4000 powder group was significantly increased compared to the constipation model group. Lactobacillus ), genus Rombutz ( Romboutsia Corynebacterium spp. Corynebacterium ), genus *Faecalibacterium* Faecalibaculum ), Mammalian cocci ( Mammaliicoccus ), Zurich bacillus ( Turicibacter Clostridium ( Clostridium The relative abundance of ) increased significantly. Lachnospiraceae _NK4A136_group、norank_f_ Oscillospiraceae Clubblerella ( Colidextribacter The relative abundance of *Romebutsiella* was significantly decreased compared to the constipation model group. In the cecal contents microbiome of the LIHUO 1978 group rats, the abundance of *Romebutsiella* spp. was significantly higher. Romboutsia Lactobacillus ( ) Lactobacillus Corynebacterium spp. Corynebacterium Staphylococcus spp. Staphylococcus ), Lactobacillus spp. Limosilactobacillus ), Zurich bacillus ( Turicibacter ), Sea cocci ( Jeotgalicoccus The relative abundance of norank_f_ increased significantly. Muribaculaceae , Christensenellaceae _R-7_group, Cerobacterium spp. ( Colidextribacter The relative abundance of *Lactobacillus* was significantly decreased in the heat-inactivated LIHUO 1978 group of rats compared to the constipation model group. Lactobacillus ), genus Rombutz ( Romboutsia Staphylococcus spp. Staphylococcus ), Mammalian cocci ( Mammaliicoccus ), Zurich bacillus ( Turicibacter The relative abundance of norank_f_ increased significantly. Muribaculaceae , Lachnospiraceae _NK4A136_group、norank_f_ Oscillospiraceae Rumenococcus ( Ruminococcus ), Clubbacterium ( Colidextribacter The relative abundance of ) decreased significantly (see Figure 26 ).

[0277] The experimental results of this embodiment show that both live and heat-inactivated *Lactobacillus acidophilus* LIHUO 1978 bacteria can improve the β-diversity of the cecal contents microbiome in constipated rats. Live *Lactobacillus acidophilus* LIHUO 1978 can significantly increase the *Lactobacillus* spp. (…) in the cecal contents microbiome of constipated rats. Lactobacillus ), Lactobacillus spp. Limosilactobacillus The relative abundance of *Lactobacillus acidophilus* LIHUO 1978 heat-inactivated cells significantly increased the abundance of *Lactobacillus* spp. in the cecal contents microbiome of constipated rats. Lactobacillus The relative abundance of ) regulates the gut microbiota.

[0278] Example 15 Effects of live and heat-inactivated Lactobacillus acidophilus LIHUO 1978 bacteria on the content of short-chain fatty acids in the intestinal contents of constipated rats

[0279] Experimental methods:

[0280] In Example 9, colon contents of 5 rats were randomly selected from each group, and Shanghai Meiji Biomedical Technology Co., Ltd. was commissioned to conduct the detection of short-chain fatty acids: isobutyric acid, isovaleric acid, and valeric acid using the GC-MC method.

[0281] The specific results are as follows:

[0282] Compared with the normal group, the concentration of isobutyric acid in the colonic contents of rats in the constipation model group decreased significantly by 54.84%. Compared with the constipation model group, the concentration of isobutyric acid in the colonic contents of rats in the polyethylene glycol 4000 powder group and the heat-inactivated LIHUO 1978 group increased by 35.71% and 42.86%, respectively, but the difference was not significant. However, the concentration of isobutyric acid in the colonic contents of rats in the LIHUO 1978 group increased significantly by 92.86% (see...). Figure 27 ).

[0283] Compared with the normal group, the isovaleric acid concentration in the colonic contents of rats in the constipation model group decreased significantly by 63.04%. Compared with the constipation model group, the isovaleric acid concentration in the colonic contents of rats in the polyethylene glycol 4000 powder group and the heat-inactivated LIHUO 1978 group increased by 82.35% and 82.35%, respectively, but the difference was not statistically significant. However, the isovaleric acid concentration in the colonic contents of rats in the LIHUO 1978 group increased significantly by 129.41% (see...). Figure 27 ).

[0284] Compared with the normal group, the valeric acid concentration in the colonic contents of rats in the constipation model group decreased significantly by 43.14%. Compared with the constipation model group, the valeric acid concentration in the colonic contents of rats in the polyethylene glycol 4000 powder group and the heat-inactivated LIHUO 1978 group increased by 31.03% and 48.28%, respectively, but the differences were not statistically significant. However, the valeric acid concentration in the colonic contents of rats in the LIHUO 1978 group increased significantly by 82.76% (see...). Figure 27 ).

[0285] Isobutyric acid, isovaleric acid, and valerate are short-chain fatty acids that can serve as energy for intestinal epithelial cells and promote mucosal growth. In this embodiment, live Lactobacillus acidophilus LIHUO 1978 can increase the concentration of isobutyric acid, isovaleric acid, and valerate in constipated rats, promote the growth of colonic tissue mucosa, reduce fecal transit time, and relieve constipation.

[0286] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A strain of Lactobacillus acidophilus ( Lactobacillus acidophilus )LIHUO 1978, characterized in that, The Lactobacillus acidophilus LIHUO 1978 described herein has the accession number GDMCC No:65514.

2. A preparation of Lactobacillus acidophilus LIHUO 1978, characterized in that, The Lactobacillus acidophilus LIHUO 1978 preparation is obtained by culturing the Lactobacillus acidophilus LIHUO 1978 of claim 1 in a culture medium; the Lactobacillus acidophilus LIHUO 1978 preparation is selected from one or more of live bacteria, fermentation broth, fermentation broth precipitate, heat-inactivated bacterial cells, and lyophilized powder.

3. A microbial inoculant, characterized in that, The microbial agent contains Lactobacillus acidophilus LIHUO 1978 as described in claim 1 or Lactobacillus acidophilus LIHUO 1978 preparation as described in claim 2.

4. The use of Lactobacillus acidophilus LIHUO 1978 as described in claim 1, or the Lactobacillus acidophilus LIHUO 1978 preparation as described in claim 2, or the microbial agent as described in claim 3, in the preparation of a medicament for treating chemotherapy-induced diarrhea.

5. The application according to claim 4, characterized in that, The viable count of Lactobacillus acidophilus LIHUO 1978 in the drug is 1.00 × 10⁻⁶. 6 -1.00×10 12 CFU / mL or 1.00×10 6 -1.00×10 12 CFU / g.

6. The application according to claim 5, characterized in that, The viable count of Lactobacillus acidophilus LIHUO 1978 in the drug is 5.00 × 10⁻⁶. 9 CFU / mL or 5.00×10 9 CFU / g.

7. The application according to any one of claims 4-6, characterized in that, The drug has at least one of the following effects: (1) Significantly reduced diarrhea scores; (2) Significantly increases the length of the small intestine; (3) Significantly reduced jejunal tissue pathology score; (4) Significantly reduced serum interleukin-1β concentration; (5) Significantly increased serum interleukin-10 concentration.

8. A drug for relieving chemotherapy-induced diarrhea, characterized in that, The drug comprises Lactobacillus acidophilus LIHUO 1978 as described in claim 1 or Lactobacillus acidophilus LIHUO as described in claim 2. The preparation of 1978 or the microbial agent as described in claim 3.

9. The use of Lactobacillus acidophilus LIHUO 1978 as described in claim 1, or the Lactobacillus acidophilus LIHUO 1978 preparation as described in claim 2, or the microbial agent as described in claim 3, in the preparation of products for improving constipation.

10. The application according to claim 9, characterized in that, The products mentioned are health foods, animal nutrition products, or pharmaceuticals.

11. The application according to claim 10, characterized in that, When the product is a pharmaceutical product, the pharmaceutical product also includes pharmaceutically acceptable excipients; When the product is a health food, the health food also includes excipients acceptable to food.

12. The application according to claim 9, characterized in that, The viable count of Lactobacillus acidophilus LIHUO 1978 in the product is 1.00 × 10⁻⁶. 6 -1.00×10 12 CFU / mL or 1.00×10 6 -1.00×10 12 CFU / g.

13. The application according to claim 12, characterized in that, The viable count of Lactobacillus acidophilus LIHUO 1978 in the product is 1.00 × 10⁻⁶. 8 CFU / mL or 1.00×10 8 CFU / g.

14. The application according to any one of claims 9-13, characterized in that, The product has at least one of the following functions: (1) Significantly increases fecal particle count and fecal moisture content; (2) Significantly improves gastrointestinal propulsion rate; (3) Significantly reduced serum endothelin-1, serum vasoactive intestinal peptide, serum somatostatin, and serum nitric oxide synthase concentrations; (4) Significantly increased serum motilin, serum substance P, serum acetylcholine, colon tissue 5-hydroxytryptamine, and colon tissue interleukin-10 concentrations; (5) Significantly increases the content of mucus-secreting goblet cells; (6) Significantly increased the concentration of aquaporin 9 in colonic tissue; (7) Regulates the intestinal flora of the cecal contents; (8) Significantly increases the concentration of isobutyric acid, isovaleric acid, and valeric acid in colon contents.

15. A product for improving constipation, characterized in that, The product includes Lactobacillus acidophilus LIHUO 1978 as described in claim 1 or Lactobacillus acidophilus LIHUO as described in claim 2. The preparation of 1978 or the microbial agent as described in claim 3.

16. The use of Lactobacillus acidophilus LIHUO 1978 as described in claim 1, or the Lactobacillus acidophilus LIHUO 1978 preparation as described in claim 2, or the microbial agent as described in claim 4, in the preparation of products for regulating intestinal flora.

17. The application according to claim 16, characterized in that, The product described can significantly increase the relative abundance of Lactobacillus and Lactobacillus mucosae in the intestine.

18. A product for regulating intestinal flora, characterized in that, The product includes Lactobacillus acidophilus LIHUO 1978 as described in claim 1 or Lactobacillus acidophilus LIHUO as described in claim 2. The preparation of 1978 or the microbial agent as described in claim 3.

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