Lactobacillus casei for warming uterus, dispelling cold and improving breast health and metaplasm thereof

Abstract

The invention discloses a lactobacillus casei strain capable of warming uterus, dispelling cold and improving breast health and a metagen thereof, and belongs to the technical field of microbial application, the lactobacillus casei strain WZA-LC26 is separated from human excrement, and the biological preservation number of the lactobacillus casei strain is CCTCC (China Center for Type Culture Collection) NO: M20241410. According to the invention, the lactobacillus casei capable of warming uterus, dispelling cold and improving breast health is screened out, aftergrowth elements and medicinal and edible substances are combined, a natural and efficient health conditioning scheme capable of treating both symptoms and root causes is created, and through the combination, the effects of warming channels, dispelling cold, promoting blood circulation, removing blood stasis, soothing liver-qi stagnation and the like of traditional Chinese medicines can be fully utilized, so that the effects of warming uterus, dispelling cold, promoting blood circulation, removing blood stasis, soothing liver-qi stagnation and the like are achieved; the curative effect and the safety of the medicine can be remarkably improved by virtue of the multi-target effects of immunoregulation, anti-inflammation and anti-oxidation, endocrine balance promotion and the like of the metaplasts.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a strain of Lactobacillus casei that warms the uterus, dispels cold and improves breast health, and a postbiotic thereof. Background Art

[0002] In recent years, due to changes in lifestyle and increased environmental pressure, women's health problems, especially those related to uterine cold and breast health, have become increasingly prominent. According to the etiology, Chinese medicine divides primary dysmenorrhea into four types: cold stagnation and blood stasis, qi stagnation and blood stasis, qi and blood deficiency, and kidney deficiency and blood stasis. Among them, cold stagnation and blood stasis is the most common clinical type. Its etiology is mostly dampness and cold during menstruation, or excessive cold food, cold invades Chongren, causing qi and blood stagnation, uterine qi and blood stagnation, and dysmenorrhea. Uterine cold may not only cause reproductive health problems such as irregular menstruation, dysmenorrhea and infertility in women, but may also further affect women's endocrine balance and overall health status. Breast hyperplasia is a benign proliferative disease caused by excessive glandular proliferation and is the most common breast disease. The incidence of breast hyperplasia is mainly middle-aged women, but in recent years it has shown a trend of younger age. Chinese medicine believes that the cause of breast hyperplasia is mainly the blockage of milk channels caused by liver qi stagnation, while modern medicine believes that the disease is caused by hormone disorders such as endocrine disorders. Patients with breast hyperplasia will feel breast pain when the disease occurs, and even small nodules can be felt. The course of the disease is prone to recurrence, and in severe cases, it may have a tendency to cause cancer, causing inconvenience and harm to the patient's life and health. Therefore, the treatment of breast hyperplasia has become a hot topic of research at home and abroad.

[0003] Modern medicine treats dysmenorrhea with non-steroidal anti-inflammatory analgesics, contraceptives, etc., which can only temporarily relieve pain, have no cure, and have many side effects. Traditional Chinese medicine can achieve overall regulation and treat both the symptoms and the root causes through the rational combination of drugs. At present, there is no good medicine to prevent breast hyperplasia. Western medicine generally uses hormone therapy to treat breast hyperplasia, mainly through oral estrogen antagonists or vitamins to reduce the estrogen content, but it often leads to other problems such as abnormal menstruation and obesity. For women with a family history of breast cancer, multiple nodules and increasing size year by year, Western medicine recommends surgical removal, but the risk of recurrence cannot be avoided. Traditional Chinese medicine has thousands of years of clinical experience in the treatment of breast diseases. Compared with Western medicine, traditional Chinese medicine has greater hope for preventing breast diseases.

[0004] In recent years, combining edible and medicinal substances with probiotics and their metabolites (postbiotics) has become an emerging direction for preventing and treating dysmenorrhea and improving breast health. This combination method provides a safe, effective and sustainable solution for women's health problems through the cross-complementarity of traditional Chinese medicine theory and modern scientific research. Edible and medicinal substances are the core content of traditional Chinese medicine diet conditioning. They can be used as drugs for disease treatment and as daily diet for health care. As a research hotspot of modern biotechnology, probiotics and their metabolites (postbiotics) further optimize the internal environment of the body by regulating the balance of intestinal flora and enhancing immune function. They also play an important role in regulating endocrine, reducing chronic inflammation, and improving metabolic function. Therefore, screening specific strains with the ability to warm the uterus and dispel cold and improve breast health, combining edible and medicinal substances with probiotics and their metabolites (postbiotics) can break the limitations of traditional treatment by treating both inside and outside, and has great development potential and application value. Summary of the invention

[0005] In view of the above-mentioned prior art, the purpose of the present invention is to provide a strain of Lactobacillus casei and its postbiotics for warming the uterus, dispelling cold and improving breast health.

[0006] To achieve the above object, the present invention adopts the following technical solution:

[0007] The first aspect of the present invention provides a strain of Lactobacillus casei WZA-LC26, which has been deposited in the China Center for Type Culture Collection (CCTCC for short, address: Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province) on July 1, 2024, and its biological preservation number is: CCTCC NO: M20241410.

[0008] A second aspect of the present invention provides a Lactobacillus casei postbiotic, wherein the Lactobacillus casei postbiotic comprises the fermentation metabolites of the Lactobacillus casei as active ingredients.

[0009] Preferably, the Lactobacillus casei postbiotic is prepared by the following method:

[0010] The Lactobacillus casei is inoculated into a fermentation medium for fermentation and culture, and then inactivated after fermentation to obtain a fermentation liquid, and the fermentation liquid is centrifuged to obtain a supernatant, and then dried to obtain a Lactobacillus casei postbiotic;

[0011] The fermentation medium is based on MRS liquid medium, and a mixture of medicine and food with a mass fraction of 4-10% is added to the basic medium;

[0012] The medicine-food mixture is composed of perilla leaves, cistanche deserticola, sea buckthorn and perilla stems in a mass ratio of (30-50): (20-40): (10-30): (5-15).

[0013] Preferably, the fermentation temperature is 30-37°C, and the fermentation time is 36-60h.

[0014] Preferably, the inoculation amount of Lactobacillus casei is 5%-15% of the volume of the fermentation medium.

[0015] The third aspect of the present invention provides the use of the above-mentioned Lactobacillus casei or Lactobacillus casei postbiotics in the preparation of products for warming the uterus and dispelling cold.

[0016] A fourth aspect of the present invention provides the use of the above-mentioned Lactobacillus casei or Lactobacillus casei postbiotics in the preparation of products for improving breast health.

[0017] The mechanism of action of the medicinal and edible substances selected by the present invention is:

[0018] In terms of warming the uterus and dispelling cold:

[0019] Perilla leaves: dispel cold and relieve exterior symptoms, promote qi circulation and relieve pain, directly act on the uterus and relieve cold and blood stasis.

[0020] Cistanche: tonifies the kidney and strengthens yang, warms the meridians and dredges the collaterals, and regulates uterine coldness and discomfort caused by kidney yang deficiency.

[0021] Seabuckthorn: Promotes blood circulation and removes blood stasis, relieves uterine cold and dysmenorrhea caused by blood stasis.

[0022] Perilla stem: Soothes the liver and regulates qi, regulating abdominal pain and menstrual disorders caused by qi stagnation.

[0023] To improve breast health:

[0024] Perilla leaves: soothe the liver and relieve depression, promote blood circulation and remove blood stasis, and relieve breast pain caused by liver qi stagnation.

[0025] Cistanche deserticola: nourishes the kidney and replenishes essence, improves endocrine balance, and reduces the hormonal imbalance that causes breast hyperplasia.

[0026] Sea buckthorn: It has strong antioxidant and anti-inflammatory properties, improves breast tissue inflammation and blood stasis, and helps to clear the milk meridians.

[0027] Perilla stem: regulates qi and relieves chest tightness, and relieves breast hyperplasia caused by qi stagnation and blood stasis.

[0028] Beneficial effects of the present invention:

[0029] 1. The present invention screens out a strain of Lactobacillus casei that can warm the uterus, dispel cold and improve breast health, and combines its postbiotics with medicinal and edible substances, innovatively and organically integrates the overall conditioning concept of traditional Chinese medicine with the action mechanism of modern probiotics and their metabolites (postbiotics), and creates a natural and efficient health conditioning program that treats both the symptoms and the root causes. Through this combination, not only can the effects of Chinese medicine such as warming the meridians and dispelling cold, promoting blood circulation and removing blood stasis, and soothing the liver and relieving depression be fully utilized, but also the multi-target effects of postbiotics such as immune regulation, anti-inflammatory and antioxidant, and promotion of endocrine balance can be used to significantly improve the efficacy and safety of the drug.

[0030] 2. The present invention adopts postbiotics instead of live bacteria, avoiding the risk of flora disorder or infection that may be caused by traditional probiotic live bacteria, while improving the stability and controllability of the product; both medicinal and edible substances are derived from natural substances, have little side effects, are suitable for long-term use, and can be used not only for symptom relief, but also for daily health management and disease prevention; in terms of preventing and treating dysmenorrhea, medicinal and edible substances can directly act on the uterus and meridians, warm the internal environment, and improve menstrual pain caused by cold coagulation and blood stasis, and qi stagnation and blood stasis, while probiotics and postbiotics can regulate intestinal flora, improve the imbalance of hormone levels in the body, and further relieve the symptoms of dysmenorrhea, and the metabolites in postbiotics can resist inflammation and regulate immune function, which helps to inhibit the inflammatory response related to dysmenorrhea, thereby achieving both symptomatic and root treatment; in terms of improving breast health, Chinese medicine and food homologous substances have the effects of soothing the liver and relieving depression, promoting blood circulation and removing blood stasis, and can directly improve the milk network obstruction caused by liver qi stagnation. In addition, probiotics and postbiotics affect the metabolism and excretion of estrogen by regulating intestinal flora, fundamentally reducing the adverse effects of endocrine disorders on breast health. DETAILED DESCRIPTION

[0031] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0032] As described in the background technology, in recent years, women's health problems, especially those related to uterine cold and breast health, have become increasingly prominent. Western medicine mostly uses non-steroidal anti-inflammatory analgesics or contraceptives to relieve dysmenorrhea, and uses hormone therapy (such as estrogen antagonists) or vitamins to regulate estrogen content to treat breast hyperplasia, but it cannot fundamentally solve the problem. Traditional Chinese medicine has an overall adjustment concept of treating both the symptoms and the root causes in the treatment of dysmenorrhea and breast hyperplasia, and has rich clinical experience. It shows greater potential for the prevention and treatment of dysmenorrhea and breast hyperplasia. However, traditional Chinese medicine ingredients have low bioavailability and slow effects, which makes it difficult to adapt to the needs of modern consumers for safety, efficiency and convenience.

[0033] Based on this, the present invention introduces modern fermentation technology on the basis of traditional Chinese medicine theory, and forms a multi-target comprehensive conditioning mechanism through the combination of postbiotics and medicinal and edible substances. Through the synergistic effect of traditional Chinese medicine and postbiotics, it can not only fully utilize the effects of traditional Chinese medicine such as warming the meridians and dispelling cold, promoting blood circulation and removing blood stasis, soothing the liver and relieving depression, but also make use of the multi-target effects of postbiotics such as immune regulation, anti-inflammatory and antioxidant, and promoting endocrine balance, thereby significantly improving the efficacy and safety of the drug.

[0034] In order to enable those skilled in the art to more clearly understand the technical solution of the present application, the technical solution of the present application will be described in detail below in conjunction with specific embodiments.

[0035] The test materials used in the examples of the present invention are all conventional test materials in the art and can be purchased through commercial channels.

[0036] Example 1: Isolation and identification of strains:

[0037] The Lacticaseibacillus casei WZA-LC26 strain of the present invention is separated and obtained from the feces of healthy adults.

[0038] 1. Isolation of strains:

[0039] Healthy adult feces were used as samples. 10 g of the sample stored in 30% (v / v) glycerol was added to 90 mL of normal saline under sterile conditions. 1 mL of the suspension was diluted 10-fold to 10 -6 . From Gradient 10 -4 -10 -6 100 μL of the sample dilution was spread on the MRS solid plate in turn, and cultured in an anaerobic incubator at 37°C for 48 h. The colonies with calcium dissolution circles were picked according to the size, color, gloss and transparency of the microbial colonies. After streaking and purification on the MRS plate three times by the streaking method, the strain was stored in a refrigerator at 4°C to obtain strain WZA-LC26.

[0040] 2. Strain identification:

[0041] The WZA-LC26 strain was sent to the Henan Industrial Microbial Strain Engineering Technology Research Center for 16SrDNA gene sequence identification, and the identification results showed that the strain belonged to Lactobacillus casei. The strain was biologically preserved, and the preservation information is as follows:

[0042] Bacterial species name: Lactobacillus casei WZA-LC26

[0043] Latin name: Lacticaseibacillus casei

[0044] Collection institution: China Center for Type Culture Collection

[0045] Abbreviation of depository institution: CCTCC

[0046] Address: Wuhan University, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province

[0047] Date of deposit: July 1, 2024

[0048] Deposit number: CCTCC NO: M20241410.

[0049] Example 2: Preparation of Lactobacillus casei WZA-LC26 bacterial agent:

[0050] Lactobacillus casei WZA-LC26 was inoculated into MRS solid medium for activation, and the activated Lactobacillus casei was inoculated into MRS liquid medium at a volume fraction of 10%, and cultured at 35°C for 48 h. Subsequently, the culture solution was centrifuged (4,000 rpm, 10 min), the supernatant was discarded, the precipitated cells were collected, and the cells were resuspended in sterile saline (0.85% NaCl) to adjust the concentration of the bacterial suspension to 1×10 8 cfu / mL.

[0051] Example 3: Preparation of Lactobacillus casei postbiotics:

[0052] (1) Preparation of fermentation medium:

[0053] Adding 7% by weight of the medicine-food mixture into the MRS liquid medium to prepare a fermentation medium;

[0054] The medicine-food mixture is prepared by the following method:

[0055] The perilla leaves, Cistanche deserticola, sea buckthorn and perilla stems are mixed evenly in a mass ratio of 4:3:2:1, and crushed to obtain a medicine-food mixture;

[0056] (2) The activated Lactobacillus casei (viable count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 10% (volume fraction), and fermentation culture was carried out at 35°C for 48h to obtain a fermentation broth;

[0057] (3) The fermentation broth was treated at 110° C. for 30 min, the supernatant was collected by centrifugation, and dried to obtain Lactobacillus casei postbiotics.

[0058] Example 4: Preparation of Lactobacillus casei postbiotics:

[0059] (1) Preparation of fermentation medium:

[0060] Adding 4% by weight of the medicine-food mixture into the MRS liquid medium to prepare a fermentation medium;

[0061] The medicine-food mixture is prepared by the following method:

[0062] The perilla leaves, Cistanche deserticola, sea buckthorn and perilla stems are mixed evenly in a mass ratio of 3:2:1:0.5, and crushed to obtain a medicine-food mixture;

[0063] (2) The activated Lactobacillus casei (viable count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 5% (volume fraction), and fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth;

[0064] (3) The fermentation broth was treated at 110° C. for 30 min, the supernatant was collected by centrifugation, and dried to obtain Lactobacillus casei postbiotics.

[0065] Example 5: Preparation of Lactobacillus casei postbiotics:

[0066] (1) Preparation of fermentation medium:

[0067] Adding 10% by weight of the medicine-food mixture into the MRS liquid culture medium to prepare a fermentation medium;

[0068] The medicine-food mixture is prepared by the following method:

[0069] Evenly mix the perilla leaves, Cistanche deserticola, seabuckthorn and perilla stems in a mass ratio of 5:4:3:1.5, and grind them to obtain a medicine-food mixture;

[0070] (2) The activated Lactobacillus casei (viable count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 15% (volume fraction), and fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth;

[0071] (3) The fermentation broth was treated at 110° C. for 30 min, the supernatant was collected by centrifugation, and dried to obtain Lactobacillus casei postbiotics.

[0072] Comparative Example 1:

[0073] (1) Preparation of fermentation medium:

[0074] Adding 7% by mass of Perilla leaf powder into MRS liquid medium to prepare a fermentation medium;

[0075] (2) The activated Lactobacillus casei (viable count 1×10 8cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 10% (volume fraction), and fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth;

[0076] (3) The fermentation broth was treated at 110° C. for 30 min, the supernatant was collected by centrifugation, and dried to obtain Lactobacillus casei postbiotics.

[0077] Comparative Example 2:

[0078] (1) Preparation of fermentation medium:

[0079] Adding 7% by mass of Cistanche deserticola powder into MRS liquid culture medium to prepare a fermentation medium;

[0080] (2) The activated Lactobacillus casei (viable count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 10% (volume fraction), and fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth;

[0081] (3) The fermentation broth was treated at 110° C. for 30 min, the supernatant was collected by centrifugation, and dried to obtain Lactobacillus casei postbiotics.

[0082] Comparative Example 3:

[0083] (1) Preparation of fermentation medium:

[0084] Adding 7% by mass of seabuckthorn powder into MRS liquid medium to prepare a fermentation medium;

[0085] (2) The activated Lactobacillus casei (viable count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 10% (volume fraction), and fermentation culture was carried out at 35°C for 48h to obtain a fermentation broth;

[0086] (3) The fermentation broth was treated at 110° C. for 30 min, the supernatant was collected by centrifugation, and dried to obtain Lactobacillus casei postbiotics.

[0087] Comparative Example 4:

[0088] (1) Preparation of fermentation medium:

[0089] Adding 7% by mass of perilla stem powder into MRS liquid medium to prepare a fermentation medium;

[0090] (2) The activated Lactobacillus casei (viable count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 10% (volume fraction), and fermentation culture was carried out at 35°C for 48h to obtain a fermentation broth;

[0091] (3) The fermentation broth was treated at 110° C. for 30 min, the supernatant was collected by centrifugation, and dried to obtain Lactobacillus casei postbiotics.

[0092] Experimental Example 1: Effects of Lactobacillus casei and its postbiotics on rats with dysmenorrhea model of cold coagulation and blood stasis type:

[0093] 1. Construction of rat model of dysmenorrhea caused by cold stagnation and blood stasis:

[0094] A rat model of dysmenorrhea with cold stagnation and blood stasis was constructed by referring to the method 1.2 in the literature "Meng Tongyu. Study on the intervention of modified Shaofu Zhuyu Decoction on the rat model of dysmenorrhea with cold stagnation and blood stasis. Feet and Health Care, 2018".

[0095] The ice water freezing method was used to immerse female rats (weighing about 200 g) in estrus in 0-4°C ice water once a day for 6 minutes for 35 consecutive days to induce a cold coagulation and congestion model. At the same time, the rats were subcutaneously injected with estradiol benzoate injection at 0.03 mg / rat / d to synchronize the uterus and create a primary dysmenorrhea model.

[0096] 2. Grouping and dosing:

[0097] The rats with dysmenorrhea model of cold coagulation and blood stasis were randomly divided into 7 groups, 8 rats in each group, for a total of 56 rats.

[0098] Drug administration began on the 15th day of modeling, and the drug administration of each treatment group was as follows:

[0099] Treatment group 1: rats were gavaged with 1.5 g / kg of the Lactobacillus casei WZA-LC26 bacterial agent prepared in Example 2;

[0100] Treatment 2: rats were gavaged with 1.5 g / kg of Lactobacillus casei postbiotics prepared in Example 3;

[0101] Treatment group 3: rats were gavaged with 1.5 g / kg of Lactobacillus casei postbiotics prepared in comparative example 1;

[0102] Treatment 4: rats were gavaged with 1.5 g / kg of Lactobacillus casei postbiotics prepared in Comparative Example 2;

[0103] Treatment 5: rats were gavaged with 1.5 g / kg of Lactobacillus casei postbiotics prepared in Comparative Example 3;

[0104] Treatment group 6: rats were gavaged with 1.5 g / kg of Lactobacillus casei postbiotics prepared in comparative example 4;

[0105] The model group was gavaged with an equal amount of normal saline; each treatment group was given medication once a day for 35 consecutive days.

[0106] 3. Test indicators:

[0107] The drugs were administered continuously for 35 days. One hour after the last day of administration, 2 U of oxytocin was injected intraperitoneally into each rat. The number of rats twisting within 30 minutes was observed, and the twisting latency and inhibition rate of the number of rats twisting were calculated.

[0108] Writhing latency = writhing start time - oxytocin injection time;

[0109] The writhing inhibition rate = (mean number of writhing in the model group - mean number of writhing in the drug-treated group) / mean number of writhing in the model group × 100%.

[0110] 4. Test results:

[0111] Table 1 The writhing reaction of rats treated with different methods

[0112]

[0113]

[0114] As can be seen from Table 1, compared with the model group, the rats fed with Lactobacillus casei WZA-LC26 bacterial agent and Lactobacillus casei postbiotics with different treatments can be prolonged to varying degrees, and the number of writhing times is also reduced. Among them, the rats fed with the Lactobacillus casei postbiotics prepared in Example 3 have the longest writhing latency and the least number of writhing times, indicating that the combination of Lactobacillus casei postbiotics and the medicine-food homologous mixture in the present application has a significant therapeutic effect on the treatment of rats with cold coagulation and blood stasis type dysmenorrhea model.

[0115] Experimental Example 2: Effects of Lactobacillus casei and its postbiotics on a rat model of mammary hyperplasia:

[0116] 1. Construction of mammary hyperplasia rat model:

[0117] A rat model was constructed by referring to the method 2.1 in the literature “Ye Yancheng, Ding Gaoheng, Ma Zhanjun, et al. Effects of Rukang Capsule on Mammary Tissue and Related Receptor Expression in Rats with Breast Hyperplasia. Chinese Journal of Traditional Chinese Medicine and Pharmacy, 2019”.

[0118] The model was established by sequential estrogen and progesterone method. Estradiol benzoate was injected into the posterior and lateral hind limbs of female non-pregnant SPF SD rats (270-300 days old, weighing 300-400g) at a body weight of 0.5 mg / (kg·d) once a day, alternating between left and right, for 25 consecutive days, followed by intraperitoneal injection of progesterone, 5 mg / (kg·d), once a day, for 5 consecutive days (body weight was measured once every 3 days, and the dosage was adjusted in time according to the change of body weight). The normal control group was injected with physiological saline intramuscularly at a dose of 0.1 mL / rat, once a day, for 30 consecutive days. Changes in related indicators of the model rats were observed and analyzed, such as poor spirits, dull hair, decreased appetite, ovarian dysfunction, breast enlargement, and proliferative lesions of breast tissue after modeling, indicating that the rat model of breast hyperplasia was successfully established.

[0119] 2. Grouping and dosing:

[0120] The successfully constructed rat model of mammary hyperplasia was randomly divided into 7 groups, namely the model group and treatment groups 1-6, with 8 rats in each group. In addition, a normal control group of 8 rats was set up, for a total of 64 rats.

[0121] The medication of each treatment group is as follows:

[0122] Treatment group 1: rats were gavaged with 2 g / kg of the Lactobacillus casei WZA-LC26 bacterial agent prepared in Example 2;

[0123] Treatment 2: rats were gavaged with 2 g / kg of Lactobacillus casei postbiotics prepared in Example 3;

[0124] Treatment group 3: rats were gavaged with 2 g / kg of Lactobacillus casei postbiotics prepared in comparative example 1;

[0125] Treatment 4: rats were gavaged with 2 g / kg of Lactobacillus casei postbiotics prepared in Comparative Example 2;

[0126] Treatment 5: gavage the rats with 2 g / kg of Lactobacillus casei postbiotics prepared in Comparative Example 3;

[0127] Treatment group 6: gavage the rats with 2 g / kg of Lactobacillus casei postbiotics prepared in comparative example 4;

[0128] The normal control group and the model group were gavaged with an equal amount of saline; each treatment group was given medication once a day for 30 consecutive days.

[0129] 3. Test indicators:

[0130] 3.1 Determination of rat nipple diameter and height:

[0131] The diameter and height (mm) of the second pair of nipples on the rat chest were accurately measured with a vernier caliper 24 hours after the last administration.

[0132] 3.2 Estradiol (E 2 ) and progesterone (P) content measurement:

[0133] 48 hours after the last administration, 10 mL of blood was collected from the apex of the rat, centrifuged at 3000 r / min for 30 min, and the supernatant was placed at 4°C for 2 h before use. 2 The detection method of the kit and the P kit instructions was used to detect E in the serum of rats in each group. 2 and the level of P.

[0134] 4. Test results:

[0135] Table 2 Diameter and height of nipples of rats in each group

[0136] Group Breast diameter (mm) Nipple height (mm) Process 1 2.03 3.05 Process 2 1.73 2.61 Process 3 1.98 2.95 Process 4 1.95 2.98 Process 5 2.04 3.08 Process 6 1.93 2.89 Model Group 2.45 3.45 Normal control group 1.67 2.53

[0137] As shown in Table 2, treatment 1 is gavaged with Lactobacillus casei WZA-LC26 bacterial agent, and the breast diameter and nipple height of rats are significantly reduced compared with the model group, indicating that the Lactobacillus casei screened by the present invention has a significant improvement effect on mammary hyperplasia. Treatment 2 groups (feeding the Lactobacillus casei postbiotics prepared by Example 3 of the present application) after treatment, the improvement amplitude of rat breast diameter and nipple height is the largest, close to the level of the normal control group, and the effect is better than that of treatment 1 directly gavage feeding Lactobacillus casei WZA-LC26 bacterial agent, indicating that Lactobacillus casei postbiotics are combined with the mixed group of medicine and food homology to mammary hyperplasia and have a more direct and more significant improvement effect. Treatment 3-treatment 6 are fed respectively with the postbiotics prepared by adding a single medicine and food homology substance to MRS liquid culture medium, compared with the model group, breast diameter and nipple height are significantly reduced, but the improvement effect is not as good as treatment 2, indicating that the present invention is compounded and added to MRS culture medium by Perilla leaf, Cistanche deserticola, Hippophae rhamnoides and Perilla stems for improving mammary hyperplasia in rats and has a synergistic effect.

[0138] Table 3 Effects of different treatments on rat E 2 , the influence of P content

[0139] Group <![CDATA[E 2 / (ng / mL)]]> P / (ng / mL) Process 1 12.244 9.569 Process 2 6.245 12.027 Process 3 9.365 10.369 Process 4 10.265 9.265 Process 5 11.635 9.754 Process 6 9.458 10.068 Model Group 18.246 5.214 Normal control group 6.325 12.368

[0140] As shown in Table 3, the serum levels of estradiol (E 2 ) level significantly increased, and the progesterone (P) content significantly decreased. The casei bacteria and its postbiotics of the present invention can reduce estradiol (E 2 ) level and increased the level of progesterone (P), and treatment 2 significantly reduced E 2 To a level close to normal, and at the same time, P is increased to a level close to normal, indicating that Example 3 of the present application, through the combination of Lactobacillus casei postbiotics and the medicine-food mixture, can regulate the metabolism of estrogen and fundamentally reduce the adverse effects of endocrine disorders on breast health.

[0141] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A strain of Lactobacillus casei WZA-LC26, characterized in that: Its biological deposit number is: CCTCC NO: M20241410.

2. A Lactobacillus casei postbiotic, characterized in that The Lactobacillus casei postbiotics use the fermentation metabolites of the Lactobacillus casei according to claim 1 as active ingredients.

3. The Lactobacillus casei postbiotic according to claim 1, characterized in that The Lactobacillus casei postbiotics are prepared by the following method: Inoculating the Lactobacillus casei described in claim 1 into a fermentation medium, performing fermentation culture, inactivating the culture after fermentation to obtain a fermentation broth, centrifuging the fermentation broth to obtain a supernatant, and drying the supernatant to obtain a Lactobacillus casei postbiotic; The fermentation medium is based on MRS liquid medium, and a mixture of medicine and food with a mass fraction of 4-10% is added to the basic medium; The medicine-food mixture is composed of perilla leaves, cistanche deserticola, sea buckthorn and perilla stems in a mass ratio of (30-50): (20-40): (10-30): (5-15).

4. The Lactobacillus casei postbiotic according to claim 3, characterized in that The fermentation temperature is 30-37°C, and the fermentation time is 36-60h.

5. The Lactobacillus casei postbiotic according to claim 3, characterized in that The inoculation amount of Lactobacillus casei is 5%-15% of the volume of the fermentation medium.

6. Application of the Lactobacillus casei described in claim 1 or the Lactobacillus casei postbiotics described in any one of claims 2-5 in preparing products for warming the uterus and dispelling cold.

7. Use of the Lactobacillus casei described in claim 1 or the Lactobacillus casei postbiotics described in any one of claims 2 to 5 in preparing a product for improving breast health.

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