Application of probiotic composition in preparation of medicine for treating intestinal inflammation

Abstract

The invention discloses an application of a probiotic composition in preparation of a medicine for treating intestinal inflammation, and the probiotic composition is prepared from a lactobacillus plantarum strain LPSG1, a bifidobacterium lactis strain HH-BA68, and / or a clostridium butyricum strain GIM1.262, and is characterized in that the probiotic composition can be used for preparing the medicine for treating the intestinal inflammation, and the probiotic composition can be used for preparing the medicine for treating the intestinal inflammation, and can be used for preparing the medicine for treating the intestinal inflammation, the medicine for treating the intestinal inflammation, the medicine for treating the intestinal inflammation, the medicine for treating the intestinal inflammation and the medicine for treating the intestinal inflammation, and the medicine for treating the intestinal inflammation, the medicine for treating the intestinal inflammation and the medicine for treating the intestinal inflammation. The invention provides a precise probiotic intervention scheme for the intestinal inflammation induced by the high-salt diet for the first time, and fills the blank that a high-salt pathology scene is not focused in the prior art. By specifically inhibiting NLRP3 inflammasome pathway core genes, the cascade reaction of HSD-inflammatory factor release is blocked, and the vicious circle of'intestinal inflammation-IBD 'is restrained from the source, so that the barrier injury of intestinal inflammation is repaired, and the local inflammatory reaction is relieved.

Description

Technical Field

[0001] The present invention belongs to the field of microbial technology, and in particular relates to an application of a probiotic composition in preparing a medicine for treating intestinal inflammation. Background Art

[0002] Chronic diseases have become a leading cause of death worldwide. Unhealthy diets high in salt, fat, and sugar contribute to increased incidence of conditions such as inflammation, cardiovascular disease, and diabetes. A high-salt diet (HSD) is a common dietary habit. Excessive salt intake can lead to dysbiosis in the gut microbiome, which in turn causes chronic inflammation in the intestinal system. HSD undoubtedly exacerbates intestinal inflammation, ultimately inducing inflammatory bowel disease (IBD). Furthermore, patients with IBD may consume higher amounts of salt than the general population. In a cross-sectional study of 67 patients by Taylor et al., salt intake was higher in both men and women with Crohn's disease than in their respective control groups, and IBD morbidity and mortality rates are also increasing.

[0003] Probiotics, as living microorganisms with clear benefits to the host, play an important role in regulating the host's intestinal flora, enhancing intestinal barrier integrity, and inhibiting the growth of pathogens. Under normal physiological conditions, commensal bacteria influence metabolism, activity, immunity, and other aspects of the body through their genes, intermediates, and metabolic activity. The intestine, as the largest digestive organ in the human body and the natural habitat of a large, dynamic bacterial community, is primarily composed of Firmicutes and Bacteroidetes. These two phyla dominate the normal intestinal flora due to their mechanisms of action. Ulcerative colitis and other colonic diseases are characterized by changes in the healthy gut, and alterations in Firmicutes and Bacteroidetes are hallmarks of intestinal flora disturbances. Furthermore, Lactobacillus plantarum and Bifidobacterium exhibit potent anti-inflammatory properties, while short-chain fatty acids (SCFAs) and butyrate produced by Clostridium butyricum can protect the mucosal barrier, alleviate local inflammation, and have potent anti-cancer activity. Accumulating evidence also demonstrates that probiotics play a key role in stimulating immunity and alleviating inflammatory mechanisms.

[0004] Intestinal inflammation is a pathological condition that is often overlooked. HSD can cause imbalances in the intestinal microbiome, promote the proliferation of pathogens, alter intestinal permeability, and damage the intestinal barrier, thereby inducing intestinal inflammation. Inflammatory bowel disease (IBD), the most devastating disease after exacerbated intestinal inflammatory infiltration, is associated with a significantly increased risk of a transition from intestinal inflammation to IBD due to recurrent inflammatory episodes. Although the broad-spectrum anti-inflammatory effects of probiotics are well-established, current research has significant limitations. First, there is a lack of intervention evaluations targeting HSD-specific inflammatory models. Second, there has been no systematic comparison of the effects of different probiotic strains (such as Lactobacillus plantarum, Bifidobacterium lactis, and Clostridium butyricum) on repairing colonic structural damage (e.g., goblet cell reduction and collagen deposition), inhibiting NLRP3 inflammasome activation, and downregulating key proinflammatory cytokines (e.g., IL-17A). Finally, a comprehensive body of evidence exists regarding the mechanisms by which probiotics regulate the HSD-NLRP3-IL-1β / IL-18 signaling axis. These limitations severely restrict the precise application of probiotics in treating intestinal diseases associated with a high-salt diet. Summary of the Invention

[0005] To solve the above technical problems, the present invention proposes the use of a probiotic composition in the preparation of a drug for treating intestinal inflammation. The present invention uses mice with intestinal inflammation induced by a high-salt diet as a model to observe the improvement effects of three probiotics, Lactobacillus plantarum, Bifidobacterium lactis, and Clostridium butyricum, on the symptoms of intestinal inflammation, providing an experimental basis for the future application of probiotics in enteral nutrition treatment of patients with intestinal inflammation.

[0006] To achieve the above object, the present invention provides a use of a probiotic composition in the preparation of a medicament for treating intestinal inflammation, characterized in that the probiotic composition comprises: Lactobacillus plantarum ( Lactiplantibacillus plantarum ) strain LPSG1, and / or, Bifidobacterium lactis ( Bifidobacterium lactis ) strain HH-BA68, and / or, Clostridium butyricum ( Clostridium butyricum ) strain GIM1.262.

[0007] Furthermore, the drug is used for: Inhibits the expression of genes in the NLRP3 inflammasome pathway, including Nlrp3 、 Asc 、 Caspase1, Nf-κb and Il-1β ; Down-regulating serum inflammatory factor levels, including TNF-α, IL-1β, IL-6, IL-17A, and IL-18; and / or, Repair pathological damage to intestinal tissue, including increasing the number of goblet cells, reducing collagen fiber deposition, and restoring colon length.

[0008] Furthermore, the viable bacterial concentration of each strain in the probiotic composition is 1×10 9 CFU / mL.

[0009] Furthermore, the dosage of the drug is 1×10 9 CFU / kg body weight, and the administration period was 28 days.

[0010] Furthermore, the intestinal inflammation is induced by a high-salt diet.

[0011] The present invention also provides a medicine for treating intestinal inflammation, comprising the above-mentioned probiotic composition and pharmaceutically acceptable excipients.

[0012] Compared with the prior art, the present invention has the following advantages and technical effects: This invention, for the first time, targets high-salt diet-induced intestinal inflammation, providing a precise probiotic intervention consisting of Lactobacillus plantarum LPSG1, Bifidobacterium lactis HH-BA68, and Clostridium butyricum GIM1.262. This approach fills a gap in existing technologies that lack a focus on high-salt pathologies. By specifically inhibiting core genes in the NLRP3 inflammasome pathway, it blocks the cascade from HSD to the release of inflammatory factors, curbing the vicious cycle of intestinal inflammation leading to IBD at the source.

[0013] The three strains in this invention show clear functional complementarity: Bifidobacterium lactis efficiently repairs the physical structure of the colon, Lactobacillus plantarum significantly improves pathological damage, and Clostridium butyricum strongly inhibits cell pyroptosis. 9 Intervention with a CFU / kg dose for 28 days simultaneously achieves intestinal barrier repair and relief of systemic inflammation, providing a targeted and actionable solution for intestinal diseases associated with a high-salt diet. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 The effect of probiotics on the weight change trend of mice induced by HSD (A) and the final weight of mice (B); Figure 2 Effects of probiotics intervention on mouse colon weight (A), mouse colon length (B) and colon index (C); Figure 3 Effects of probiotic intervention on pathological changes in mouse colon tissue (A), colon fibrosis (B), and colon epithelial goblet cells (C); black arrows point to goblet cells, which are stained blue. Figure 4 Effects of probiotics intervention on the levels of serum inflammatory factors TNF-α (A), IL-1β (B), IL-18 (C), IL-6 (D) and IL-17A (E); Figure 5Effects of probiotic intervention on NLRP3 inflammasome-related factors in colon tissue Nlrp3 (A) Asc (B) Caspase1 (C) Nf-κb (D) and Il-1β (E) Effects of mRNA expression levels; In the above figures, different letters indicate significant differences between the two groups ( P <0.05). DETAILED DESCRIPTION

[0015] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0016] All raw materials of the present invention are not particularly limited in their sources and can be purchased from the market or prepared according to conventional methods well known to those skilled in the art.

[0017] There is no particular limitation on the purity of all raw materials in the present invention. The present invention preferably uses analytically pure raw materials or raw materials with a purity commonly used in the field of chemical synthesis.

[0018] 1. Materials and Methods 1.1 Materials and Reagents Lactiplantibacillus plantarum (LPSG1) was isolated and preserved by our research group, with the deposit number CGMCC No. 31425 and the deposit date of July 22, 2024, at the General Microbiology Center of the China Culture Collection Administration Committee of Microorganisms. Bifidobacterium lactis (HH-BA68) was obtained from Zhengzhou Hehe Bioengineering Technology Co., Ltd. Clostridium butyricum (GIM1.262) was obtained from the Culture Collection Center of Guangdong Institute of Microbiology. AIN-93M feed containing 8% NaCl was purchased from Changzhou Shuyi Shuer Biotechnology Co., Ltd. 0.05% saline and ELASA kit were obtained from Nanjing Jiancheng Bioengineering Institute. Alcian blue staining kit (BP-DL241) was purchased from Nanjing Senbeijia Biotechnology Co., Ltd. Hematoxylin and eosin (H&E) staining kit (C0105), BCA protein concentration kit (P0012), and 5X SDS-PAGE protein loading buffer (P0286-15 mL) was purchased from Shanghai Biotech Biotechnology Co., Ltd.; TRIzol total RNA extraction kit (R1200-50T) was purchased from Beijing Solebao Biotechnology Co., Ltd.; GlpBio SYBR Green qPCR master mix (GC26360) was purchased from Shanghai Hongye Biotechnology Co., Ltd.

[0019] 1.2 Instruments and Equipment Electronic analytical balance, Shanghai Tianmei Balance Instrument Co., Ltd., precision: 0.0001 g; Eppendorf 5424R refrigerated centrifuge, Eppendorf, Germany; multifunctional microplate reader, Meigu Molecular Instrument (Shanghai) Co., Ltd.; Leica DM4B microscope, Leica Instrument Co., Ltd., Germany; KZ-II high-speed tissue grinder, Wuhan Sewell Biotechnology Co., Ltd.; Quant StudioTM 6 Flex PCR instrument, Thermo Scientific Technology Co., Ltd., USA.

[0020] 1.3 Experimental animals Thirty-five male SPF-grade clean mice (C57BL / 6J, 6 weeks old, weighing 15–19 g) were purchased from Zhejiang Weitonglihua Laboratory Animal Technology Co., Ltd. (Laboratory Animal Production License No. SCXK (Zhejiang) 2020-0002). They were housed in the SPF animal facility of Guilin Medical College (Laboratory Unit License No. SYXK (Guizhou) 2020-0005). The animal housing environment was maintained at a temperature of (22 ± 3)°C and a relative humidity of 55%–60%. The experimental animals were kept under a light cycle of 14 h per day and well-ventilated. Animals had free access to food and clean drinking water. This study was approved by the Laboratory Animal Ethics Committee of Guilin Medical College (GLMC-IACUC-20241019).

[0021] 1.4 Probiotic culture and bacterial suspension preparation The suspensions of Lactobacillus plantarum and Bifidobacterium lactis were prepared according to the method of Li Na (Li Na, Yang Jianghua, Wei Yutuo. Optimization of high-density fermentation conditions of Lactobacillus plantarum ZJ316 [J]. China Brewing, 2021, 40(05): 43-48.), and the suspension of Clostridium butyricum was prepared according to the method of Wang Lesheng (Wang Lesheng, Li Ke, Fang Shasha, et al. Optimization of high-density fermentation and spore production conditions of Clostridium butyricum [J]. Food and Fermentation Industries, 2024: 1-11.). Lactobacillus plantarum and Bifidobacterium lactis were inoculated into MRS liquid culture medium, and Clostridium butyricum was inoculated into RCM proliferation medium. After incubation at 37 °C for 24 h under anaerobic conditions, the bacteria were counted. According to the measured viable cell count, a certain amount of Lactobacillus plantarum, Bifidobacterium lactis and Clostridium butyricum were resuspended in physiological saline to a concentration of 1×10 9 CFU / mL of bacterial suspension.

[0022] 1.5 Experimental methods 1.5.1 Animal modeling, grouping, and intervention The establishment of high salt mouse model refers to Yazawa 0et al. (Yazawa H, Miyachi M, Furukawa M, et al. Angiotensin-Converting Enzyme Inhibition Promotes Coronary Angiogenesis in the Failing Heart of Dahl Salt-Sensitive Hypertensive Rats [J]. Journal of Cardiac Failure, 2011, 17(12): 1041-50.) A total of 35 C57 male mice were randomly divided into 7 groups after one week of adaptive feeding: control group, HSD group, HSD + Lactobacillus plantarum group, HSD + Bifidobacterium lactis group, and HSD + Clostridium butyricum group. The control group was fed with AIN-93M normal feed and gavage with normal saline (1 mL / d). The other groups were fed with AIN-93M feed containing 8% NaCl and gavage with 1 mL of NaCl 100 mg / (kg·BW·d) and free access to food. At the same time, the Lactobacillus plantarum group, the Bifidobacterium lactis group, and the Clostridium butyricum group were gavage with the same amount of the corresponding live bacterial suspension (1 mL / d, 1×10 9 CFU / mL). There was no significant difference in the initial body weight of mice among the groups ( P >0.05), the animals were gavage-fed at a fixed time every day for 28 consecutive days. During the experiment, the animals were free to eat and drink, and their body weight was measured every 3 days.

[0023] 1.5.2 Experimental Animal Body Mass Measurement, Biological Sample Collection and Storage Throughout the experimental period, mice were measured and recorded at fixed times every three days. After the experimental intervention, mice were fasted for 12 hours (with free water access). They were anesthetized with ether and wiped with alcohol cotton balls. The mice were then moved to a workbench, the abdominal cavity opened, and blood samples were collected from the central abdominal artery. Blood samples were collected into pyrogen- and endotoxin-free centrifuge tubes. The samples were centrifuged at 3500 rpm at 4°C for 15 minutes, and the supernatant was quickly centrifuged and stored at -80°C. After blood sampling, the animals were placed on ice, and organs (heart, liver, kidney, and colon) were quickly dissected. These organs were washed with saline and dried with filter paper. The ratio of colon weight to colon length was calculated, and colon length was measured and photographed. A portion of colon tissue was fixed with 4% paraformaldehyde for subsequent pathological analysis. A portion was stored in TRIzol at -80°C for subsequent qRT-PCR analysis, while the remaining tissue was directly stored in a centrifuge tube at -80°C until further use.

[0024] 1.5.3 Calculation of Colon Index After colon removal, flush the feces with saline. After cleaning, weigh the feces using an electronic analytical balance. Colon index = colon mass / mouse mass.

[0025] 1.5.4 Determination of serum biochemical indicators Thaw serum samples stored at -80°C and detect the ELISA method in strict accordance with the kit instructions to detect the levels of TNF-α, IL-1β, IL-18, IL-6, and IL-17A in mouse serum (unit: ng / L).

[0026] 1.5.5 Hematoxylin-eosin and Alcian blue staining of colon tissue Paraffin sections were prepared from colon tissue samples fixed with 4% paraformaldehyde. The sections were placed in an oven at 65°C to melt the wax, dewaxed with xylene, and dehydrated with graded alcohols. Conventional hematoxylin-eosin (HE), Masson's trichrome (Masson), and Alcian blue staining were performed according to the kit protocol. Xylene I and xylene II were used for transparency. Images were acquired using a Leica DM4B optical microscope, and histopathological changes were observed at 100x magnification.

[0027] 1.5.6 Detection of mRNA expression of NLRP3-related factors in colonic inflammasomes by qRT-PCR After cleaning blood stains from an appropriate amount of colon tissue, extract the Total RNA from the tissue according to the requirements of TRIzol reagent. The concentration of the purified Total RNA was detected by UV spectrometry for subsequent experiments. Take 5μg of Total RNA, 4ul of 5xFaseting-RTSuperMix, and 11ul of RNase-Free ddH2O in a centrifuge tube and reverse transcribe them into cDNA. Take an appropriate amount of cDNA (1μL) and use qRT-PCR to detect the expression levels of related factors. In the total reaction system (10 μL): 0.2μL each of upstream and downstream primers (10 μmol / L), 2×SYBR Master mix (5 μL), Reference Dye (0.2 μL) and RNase-Free ddH2O (3.4 μL) were added, mixed thoroughly and placed in Quant Studio TMAmplification reactions were performed in a 6-Flex PCR instrument, with triplicate wells set up for each sample. A two-step amplification protocol was used: pre-denaturation at 95°C for 10 minutes, denaturation at 95°C for 15 seconds, and annealing at 60°C for 60 seconds, for 35–40 cycles. Each gene cDNA sample was amplified three times in parallel, and the average Ct value was calculated to calculate the target gene expression level according to the following formula.

[0028]

[0029] 1.6 Data Statistics and Analysis The data were statistically analyzed using the one-way ANOVA method of SPSS 23.0 statistical software. The experimental results were expressed as the mean plus or minus the standard deviation ( ±s). The LSD method was used to compare the means between the groups, and the test level was α = 0.05. P The value of <0.05 was considered statistically significant. GraphPad prism9 software was used to analyze the data.

[0030] 2. Results and Discussion 2.1 Effects of different probiotic interventions on body weight in mice induced by a high-salt diet like Figure 1 As shown in Figure 2, at the beginning of the experiment, there was no significant difference in body weight among the groups ( P >0.05). Compared with the normal group, the weight loss of mice treated with high-salt diet was more significant ( P <0.05). At the same time as the model was established, the other groups were given different probiotic suspensions for gavage intervention. The results showed that, compared with the model group, after the 9th day of the experiment, the body weight of the three probiotic intervention groups showed a recovery trend, with Bifidobacterium lactis showing the most significant recovery trend. At the end of the experiment, the final body weight of the mice in the Lactobacillus plantarum and Bifidobacterium lactis groups was significantly different from that of the HSD group ( P <0.05), the final body weight of mice in the Clostridium butyricum group had no significant difference with that in the HSD group ( P >0.05). The results suggest that probiotic intervention may restore the weight loss of mice induced by a high-salt diet by improving the inflammatory environment.

[0031] Effects of different probiotic interventions on colon length, weight, and colon index in mice fed a high-salt diet like Figure 2 Compared with the model group, there was no significant difference in the colon weight of mice in the Lactobacillus plantarum group ( P >0.05), there was a significant difference in colon length ( P <0.05), and there was no significant difference in colon index (P >0.05). There was a significant difference in colon weight in the mice in the Bifidobacterium lactis group ( P <0.05), there was a significant difference in colon length ( P <0.05), the colon index showed significant differences ( P <0.05). There was no significant difference in colon weight between the mice in the Clostridium butyricum group ( P >0.05), there was no significant difference in colon length ( P >0.05), there was no significant difference in colon index ( P >0.05). The above results indicate that Lactobacillus plantarum (to 67.00 mm) and Bifidobacterium lactis (to 69.00 mm) can restore the shortening of the colon in the high salt-induced mouse model, and the Bifidobacterium lactis group can also increase the colon weight loss caused by the model group (102.60 mg).

[0032] 2.3 Effects of different probiotic interventions on intestinal damage induced by a high-salt diet in mice The results of HE, Masson and Alcian blue staining of colon in each group are as follows Figure 3 As shown, observation of tissue sections revealed abundant and neatly arranged intestinal villi in the control group (Figure A). Glandular cells in the mucosal layer were neatly arranged, and inflammation and thickening of the muscularis propria were not observed. In contrast, mice in the HSD group exhibited significant pathological changes: altered villus morphology, disorganized intestinal glandular cells, widespread epithelial cell damage, loss of colonic crypt structure, infiltration of inflammatory cells, and significant thickening of the muscularis propria, indicating a pronounced inflammatory response. Masson staining (Figure B) revealed that compared with the control group, the muscularis propria of the HSD group was highly fibrotic, with abnormal infiltration of collagen fibers into the villi. Alcian blue staining (Figure C) revealed that goblet cells in the colon of the HSD group were significantly reduced and scattered throughout the villi. Probiotic treatment improved the colonic architecture of the mice, with normalization of the intestinal epithelium, restoration of the intestinal glandular cell and crypt structure, and reduction of inflammatory infiltration. The degree of colonic fibrosis was reduced compared to the HSD group, as evidenced by decreased collagen fiber deposition and smaller fibrotic areas. Alcian blue staining more clearly demonstrated the restoration of goblet cell numbers. These results suggest that Lactobacillus plantarum, Bifidobacterium lactis, and Clostridium butyricum can improve intestinal mucosal function, inflammatory infiltration of the muscularis, promote intestinal mucosal repair, and enhance intestinal barrier function.

[0033] 2.4 Effects of different probiotic interventions on the levels of inflammatory factors in the serum of mice induced by a high-salt diet The occurrence of intestinal inflammation is caused by a variety of pathogenic mechanisms. A high-salt diet acts on the intestinal flora, intestinal barrier, immune system, etc., driving various inflammatory mechanisms, causing abnormal activation of the immune system and triggering intestinal inflammation. Cytokines are closely related to immune disorders, mainly achieving inflammatory responses by upregulating pro-inflammatory factors and downregulating anti-inflammatory factors, and releasing inflammatory factors, mainly including: TNF-α, IL-1β, IL-18, IL-6, and IL-17A. Among them, TNF-α is the key factor causing pathological reactions, and can produce a synergistic effect with IL-1β. IL-18 and IL-6 can stimulate T and B cells to participate in inflammatory responses, while IL-17A can directly cause inflammatory damage to the intestinal mucosa and trigger local inflammation. Such as Figure 4 As shown, compared with the Control group, the levels of TNF-α, IL-1β, IL-18, IL-6, and IL-17A in the serum of mice in the HSD group were significantly increased ( P <0.05). Compared with the HSD group, the levels of TNF-α, IL-1β, IL-18, IL-6, and IL-17A in the Lactobacillus plantarum group decreased by 10.16%, 11.10%, 11.74%, 19.03%, and 41.69%, respectively, and all of these differences were statistically significant ( P <0.05); in the Bifidobacterium lactis group, TNF-α decreased by 7.10%, IL-1β decreased by 7.80%, IL-18 decreased by 12.09%, IL-6 decreased by 24.00%, and IL-17A decreased by 41.49%, and all of them were significantly different ( P <0.05); in the Clostridium butyricum group, TNF-α decreased by 11.82%, IL-1β decreased by 7.46%, IL-18 decreased by 11.23%, IL-6 decreased by 10.88%, and IL-17A decreased by 28.60%, and all of these differences were significant ( P <0.05). The above results indicate that Lactobacillus plantarum, Bifidobacterium lactis, and Clostridium butyricum can significantly downregulate the expression of inflammatory cytokines in serum and alleviate the infiltration of inflammatory cells.

[0034] 2.5 Effects of different probiotic interventions on colon tissue of mice induced by high salt diet Nlrp3 、 Asc 、 Caspase1 、 Nf-κb and Il-1β Effects on mRNA expression The activation of NLRP3 inflammasome is one of the important pathological factors leading to intestinal inflammation and is the initiator of many inflammatory pathways. The stable expression of intestinal inflammasome NLRP3 plays a very important role in maintaining the body's homeostasis. Regulating the level of NF-κB in the colon can effectively inhibit the pro-inflammatory effect of the NLRP3 pathway in the intestine and reduce the occurrence of intestinal inflammation. One of the classic signaling pathways for activating NLRP3 is: Toll-like receptor 4 agonists produce NLRP3 initiation signals through the NF-κB signaling pathway. After HSD activates the inflammasome NLRP3, it recruits and activates Caspase1 together with Asc. Nlrp3 combines with the recruited Asc and Caspase1 to form an inflammasome. At the same time, the NLRP3 inflammasome activated by NF-κB can also catalyze the maturation of downstream pro-inflammatory cytokines IL-1β and IL-18, and the colon tissue secretes pro-inflammatory factors, thereby aggravating the body's local inflammatory response. Therefore, detection Nlrp3 、 Asc 、 Caspase1 、 Nf-κb 、 Il-1β The expression of probiotics in colon tissue was investigated to explore the mechanism of the effect of different probiotic interventions on the NLRP3 signaling pathway in the colon of mice fed a high-salt diet. Figure 5 It can be seen that compared with the Control group, the HSD group mice had Nlrp3 、 Asc 、 Caspase1 、 Nf-κb 、 Il-1β The levels were significantly increased ( P <0.05), compared with the HSD group, the Lactobacillus plantarum group Nlrp3 A decrease of 85.12%, Asc decreased by 86.32%, Caspase1 A decrease of 50.01%, Nf-κb A decrease of 63.45%, Il-1β decreased by 76.35%, and there were significant differences ( P <0.05); Bifidobacterium lactis group Nlrp3 A decrease of 81.96%, Asc decreased by 69.96%, Caspase1 A decrease of 31.38%, Nf-κb A decrease of 86.13%, Il-1β decreased by 68.89%, and there were significant differences ( P <0.05); Clostridium butyricum group Nlrp3 A decrease of 72.68%, Asc decreased by 71.52%, Caspase1 A decrease of 60.87%, Nf-κb A decrease of 61.79%, Il-1β decreased by 59.96%, and there were significant differences (P <0.05). The above results show that Lactobacillus plantarum, Bifidobacterium lactis, and Clostridium butyricum can significantly reduce the expression of NLRP3 in the mouse colon inflammasome.

[0035] 3. Conclusion In summary, Lactobacillus plantarum, Bifidobacterium lactis, and Clostridium butyricum can alleviate the weight loss and colon mass loss, colon shortening, and repair inflammatory damage such as intestinal damage, colon fibrosis, and goblet cell number in mice with intestinal inflammation induced by a high-salt diet to a certain extent, thereby directly or indirectly repairing the intestinal barrier function. The degree of improvement varies. Among them, Lactobacillus plantarum has an advantage in colon pathological damage, while Bifidobacterium lactis has an advantage in colon mass and length. In addition, probiotics can also downregulate the content of inflammatory cytokines in mouse serum, restore immune function, and inhibit NLRP3 inflammasome-related factors. Nlrp3, Asc, Caspase1, Nf-κb, Il-1β This study shows that probiotics can repair the barrier damage caused by intestinal inflammation and alleviate local inflammatory responses through direct or indirect effects.

[0036] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art shall fall within the scope of protection defined by the claims of the present invention.

Claims

1. Use of a probiotic composition in preparing a medicament for treating intestinal inflammation, characterized in that: The probiotic composition comprises: Lactobacillus plantarum ( Lactiplantibacillus plantarum ) strain LPSG1, and / or, Bifidobacterium lactis ( Bifidobacterium lactis ) strain HH-BA68, and / or, choosinobacterium ( Clostridium butyricum ) strain GIM1.

262.

2. The use according to claim 1, characterized in that The drug is used to: Inhibits the expression of genes in the NLRP3 inflammasome pathway, including Nlrp3 、 Asc 、 Caspase1、Nf-κb and Il-1 β ; Down-regulating serum inflammatory factor levels, including TNF-α, IL-1β, IL-6, IL-17A, and IL-18; and / or, Repair pathological damage to intestinal tissue, including increasing the number of goblet cells, reducing collagen fiber deposition, and restoring colon length.

3. The use according to claim 1, characterized in that The concentration of live bacteria of each strain in the probiotic composition is 1×10 9 CFU / mL.

4. The use according to claim 1, characterized in that The dosage of the drug is 1×10 9 CFU / kg body weight, and the administration period was 28 days.

5. The use according to claim 1, characterized in that The intestinal inflammation is induced by a high-salt diet.

6. A drug for treating intestinal inflammation, characterized in that: The invention comprises the probiotic composition according to claim 1 and pharmaceutically acceptable excipients.

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