A strain of Lactobacillus casei for warming the uterus, dispelling cold and improving breast health and its postbiotics

Abstract

The present invention discloses a strain of Lactobacillus casei for warming the uterus and dispelling cold and improving mammary gland health and its postbiotics, belonging to the field of microbial application technology. The present invention isolates a strain of Lactobacillus casei (Lacticaseibacillus casei) WZA-LC26 from human feces, and its biological preservation number is: CCTCC NO: M20241410. The present invention screens out a strain of Lactobacillus casei for warming the uterus and dispelling cold and improving mammary gland health, and combines its postbiotics with medicinal and edible substances, creating 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 promoting endocrine balance can be used to significantly improve the efficacy and safety of the drug.

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 mammary gland health, and a postbiotic thereof. Background Art

[0002] In recent years, due to changing lifestyles and increasing environmental pressures, women's health issues, particularly those related to uterine coldness and breast health, have become increasingly prominent. Traditional Chinese Medicine (TCM) categorizes primary dysmenorrhea into four types based on etiology: cold stagnation and blood stasis, qi stagnation and blood stasis, qi and blood deficiency, and kidney deficiency and blood stasis. Cold stagnation and blood stasis is the most common clinical type, often caused by exposure to dampness and cold during menstruation, or excessive consumption of cold foods. Cold invades the Chong and Ren meridians, leading to stagnation of qi and blood, and poor uterine qi and blood flow, causing dysmenorrhea. Uterine coldness can not only lead to reproductive health issues such as irregular menstruation, dysmenorrhea, and infertility, but can also further impact a woman's endocrine balance and overall health. Fibrocystic breast disease, a benign proliferative disorder caused by glandular hyperplasia, is the most common breast disease. Fibrocystic breast disease primarily affects middle-aged women, but has recently shown a trend toward younger women. Traditional Chinese Medicine (TCM) believes that the primary cause of fibrocystic breast disease is liver qi stagnation, resulting in blocked milk channels, while modern medicine believes it is caused by hormonal imbalances, including endocrine disorders. Patients with breast hyperplasia experience breast pain and may even feel small nodules when the disease occurs. The disease is prone to recurrence and can even cause cancer in severe cases, causing inconvenience and harm to the patient's life and health. Therefore, the treatment of breast hyperplasia has become a hot topic of research both at home and abroad.

[0003] Modern medicine often uses nonsteroidal anti-inflammatory analgesics and birth control pills to treat dysmenorrhea, which only temporarily relieve pain without curing the underlying condition and often have numerous side effects. Traditional Chinese medicine, through the rational combination of drugs, can achieve overall regulation and address both the symptoms and the root cause. Currently, there is no effective medication for preventing breast hyperplasia. Western medicine generally uses hormone therapy, primarily through oral estrogen antagonists or vitamins to lower estrogen levels, but this often leads to other problems such as menstrual abnormalities and obesity. For women with a family history of breast cancer and multiple nodules that are growing larger year by year, Western medicine recommends surgical removal, but the risk of recurrence cannot be avoided.

[0004] In recent years, combining edible and medicinal substances with probiotics and their metabolites (postbiotics) has become an emerging approach for preventing and treating dysmenorrhea and improving breast health. This combined approach, through the cross-fertilization of Traditional Chinese Medicine (TCM) theory and modern scientific research, offers a safe, effective, and sustainable solution to women's health issues. Medicinal and edible substances are a core component of TCM dietary conditioning, serving both as medications for disease treatment and as part of daily diets for health and wellness. Probiotics and their metabolites (postbiotics), a research hotspot in modern biotechnology, further optimize the body's internal environment by regulating intestinal flora balance and enhancing immune function. They also play a significant role in regulating endocrine function, reducing chronic inflammation, and improving metabolic function. Therefore, screening for specific strains with the potential to warm the uterus, dispel cold, and improve breast health, and combining edible and medicinal substances with probiotics and their metabolites (postbiotics), can overcome the limitations of traditional treatments by addressing both internal and external needs, offering significant potential for development and application. 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 that can warm the uterus, dispel cold and improve mammary gland health.

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

[0007] In a first aspect of the present invention, a strain of Lactobacillus casei WZA-LC26 is provided, 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 a fermentation metabolite of the Lactobacillus casei as an active ingredient.

[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 is inactivated after fermentation to obtain a fermentation broth, and the fermentation broth is centrifuged to obtain a supernatant, which is dried to obtain a Lactobacillus casei postbiotic;

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

[0012] The medicine-food mixture consists 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 36-60 h.

[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 that improve breast health.

[0017] The mechanism of action of the medicinal and edible substances selected in 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 and relieve pain, directly act on the uterus, and relieve cold and blood stasis.

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

[0021] Sea buckthorn: 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 irregular menstruation caused by qi stagnation.

[0023] To improve breast health:

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

[0025] Cistanche: Nourishes the kidney and replenishes essence, improves endocrine balance, and reduces hormonal imbalances that are the cause of 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 warms the uterus, dispels cold, and improves breast health, and combines its postbiotics with medicinal and edible substances, innovatively integrating the holistic conditioning concept of traditional Chinese medicine with the mechanism of action of modern probiotics and their metabolites (postbiotics), creating 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 relieving liver 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 uses postbiotics instead of live bacteria, avoiding the risk of bacterial flora disorder or infection caused by traditional live probiotics, while improving the stability and controllability of the product; the medicinal and edible substances are all 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, the 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 regulate the intestinal flora, improve the imbalance of hormone levels in the body, and further relieve the symptoms of dysmenorrhea. The metabolites in postbiotics can be anti-inflammatory 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 edible substances have the effects of soothing the liver and relieving depression, promoting blood circulation and removing blood stasis, and can directly improve the milk channel obstruction caused by liver qi stagnation. In addition, probiotics and postbiotics affect the metabolism and excretion of estrogen by regulating the 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 intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present application belongs.

[0032] As described in the background art, in recent years, women's health problems, especially those related to uterine cold and breast health, have become increasingly prominent. Western medicine often 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 a holistic regulatory 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 effect, 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 dispersing cold, promoting blood circulation and removing blood stasis, and relieving liver depression, but also make use of the multi-target effects of postbiotics such as immune regulation, anti-inflammatory and antioxidant, and promotion of 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 with reference to 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 from the feces of healthy adults.

[0038] 1. Isolation of strains:

[0039] Healthy adult feces were used as samples. 10 g of the sample was stored in 30% (v / v) glycerol and 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 MRS solid plates 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 plates three times, 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 Engineering Technology Research Center for 16S rDNA gene sequence identification, which showed that the strain belonged to Lactobacillus casei. The strain was then placed on a biological deposit, and the deposit information is as follows:

[0042] Strain name: Lactobacillus casei WZA-LC26

[0043] Latin name: Lacticaseibacillus casei

[0044] Depository: 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] Deposit date: 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. 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 bacterial suspension concentration to 1×10 8 cfu / mL.

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

[0052] (1) Preparation of fermentation medium:

[0053] Adding 7% by mass of the medicinal and edible mixture to the MRS liquid culture 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 were mixed uniformly in a mass ratio of 4:3:2:1, and crushed to obtain a medicinal and edible mixture;

[0056] (2) The activated Lactobacillus casei (viable bacteria count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum size of 10% (volume fraction), and fermentation culture was carried out at 35°C for 48 hours 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 the Lactobacillus casei postbiotics.

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

[0059] (1) Preparation of fermentation medium:

[0060] Adding 4% by weight of the medicinal and edible mixture to the MRS liquid culture 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 were mixed uniformly in a mass ratio of 3:2:1:0.5, and crushed to obtain a medicinal and edible mixture;

[0063] (2) The activated Lactobacillus casei (viable bacteria count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum size of 5% (volume fraction), and the 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 the Lactobacillus casei postbiotics.

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

[0066] (1) Preparation of fermentation medium:

[0067] Adding 10% by weight of the medicinal and edible mixture to the MRS liquid culture medium to prepare a fermentation medium;

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

[0069] The perilla leaves, Cistanche deserticola, sea buckthorn and perilla stems were mixed uniformly in a mass ratio of 5:4:3:1.5, and crushed to obtain a medicinal and edible mixture;

[0070] (2) The activated Lactobacillus casei (viable bacteria count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum size 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 the Lactobacillus casei postbiotics.

[0072] Comparative Example 1:

[0073] (1) Preparation of fermentation medium:

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

[0075] (2) The activated Lactobacillus casei (viable bacteria count 1×10 8cfu / mL) was inoculated into the fermentation medium at an inoculum size 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 the Lactobacillus casei postbiotics.

[0077] Comparative Example 2:

[0078] (1) Preparation of fermentation medium:

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

[0080] (2) The activated Lactobacillus casei (viable bacteria count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum size 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 the Lactobacillus casei postbiotics.

[0082] Comparative Example 3:

[0083] (1) Preparation of fermentation medium:

[0084] adding 7% by mass of sea buckthorn powder to MRS liquid culture medium to prepare a fermentation medium;

[0085] (2) The activated Lactobacillus casei (viable bacteria count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum size of 10% (volume fraction), and fermentation culture was carried out at 35°C for 48 hours 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 the Lactobacillus casei postbiotics.

[0087] Comparative Example 4:

[0088] (1) Preparation of fermentation medium:

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

[0090] (2) The activated Lactobacillus casei (viable bacteria count 1×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum size of 10% (volume fraction), and fermentation culture was carried out at 35°C for 48 hours 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 the 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:

[0093] 1. Construction of rat model of dysmenorrhea caused by cold coagulation 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. Female rats (weighing about 200 g) in estrus were immersed in 0-4°C ice water once a day for 6 minutes each time for 35 consecutive days to induce a cold coagulation and blood stasis model by cold stimulation. At the same time, the rats were subcutaneously injected with estradiol benzoate injection 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 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 group 2: rats were gavaged with 1.5 g / kg of the Lactobacillus casei postbiotics prepared in Example 3;

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

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

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

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

[0105] The model group was gavage-fed with an equal volume 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 for 35 consecutive days. One hour after the last day of administration, 2 U of oxytocin was injected intraperitoneally into each rat. The number of writhing times of the rats was observed within 30 minutes, and the writhing latency and writhing inhibition rate were calculated.

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

[0109] 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 Writhing reactions 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 prolong the duration of writhing 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 mixture in this 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 according 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 mammary hyperplasia. Chinese Journal of Traditional Chinese Medicine and Pharmacy, 2019".

[0118] A model was established using a sequential estrogen and progesterone regimen. Non-pregnant SPF Sprague-Dawley rats (270-300 days old, weighing 300-400 g) were injected intramuscularly with estradiol benzoate (0.5 mg / kg / day) once daily, alternating between the left and right limbs, for 25 consecutive days. This was followed by an intraperitoneal injection of progesterone (5 mg / kg / day) once daily for 5 consecutive days. (Body weight was measured every 3 days, and the dosage was adjusted based on changes in body weight.) A control group received intramuscular injections of 0.1 mL / rat of normal saline once daily for 30 consecutive days. Changes in relevant parameters of the model rats were observed and analyzed. The rats exhibited poor spirits, dull fur, decreased appetite, ovarian dysfunction, breast enlargement, and hyperplastic mammary tissue, indicating the successful establishment of a rat model of mammary hyperplasia.

[0119] 2. Grouping and dosing:

[0120] The successfully constructed mammary hyperplasia rat model 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 drug administration 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 group 2: rats were gavaged with 2 g / kg of the Lactobacillus casei postbiotics prepared in Example 3;

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

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

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

[0127] Treatment group 6: rats were gavaged with 2 g / kg of the Lactobacillus casei postbiotics prepared in Comparative Example 4;

[0128] The normal control group and the model group were gavage-fed with equal amounts of normal 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 Measurement of estradiol (E2) and progesterone (P) content:

[0133] 48 hours after the last administration, 10 mL of blood was collected from the apex of the heart and centrifuged at 3000 rpm for 30 minutes. The supernatant was stored at 4°C for 2 hours before use. The levels of E2 and P in the serum of rats in each group were measured according to the instructions of the E2 and P kits.

[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, process 1 is gavaged and fed with lactobacillus casei WZA-LC26 inoculum, and the breast diameter of rat and nipple height are significantly reduced compared to model group, illustrating that the lactobacillus casei screened by the present invention has obvious improvement effect on mammary gland hyperplasia.Process 2 groups (feeding the lactobacillus casei postbiotics prepared by embodiment 3 of the application) after treatment, and rat breast diameter and nipple height improvement amplitude are maximum, close to the level of normal control group, and effect is better than process 1 and directly gavage and feed lactobacillus casei WZA-LC26 inoculum, shows that lactobacillus casei postbiotics is combined with medicine-food homologous mixed group and has more direct, more significant improvement effect on mammary gland hyperplasia.Process 3-process 6 are fed respectively with the postbiotics prepared by adding single medicine-food homologous material in MRS liquid nutrient medium, compared with model group, breast diameter and nipple height obviously decline, but improvement effect is all not as good as process 2, illustrate that the present invention is added into MRS nutrient medium by the composite of Perilla Leaf, Cistanche Deserticola, Hippophae Rhamnoides and Perilla Stem and fermented for improving rat mammary gland hyperplasia and has synergistic effect.

[0138] Table 3 Effects of different treatments on E2 and P content in rats

[0139] Group <![CDATA[E2 / (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 can be seen from Table 3, the estradiol (E2) level in the serum of the model group rats was significantly increased and the progesterone (P) content was significantly reduced. The Lactobacillus casei and its postbiotics of the present invention can reduce the estradiol (E2) level and increase the progesterone (P) level. Among them, the treatment of group 2 significantly reduced E2 to a level close to normal, while increasing P to a level close to normal. This shows that Example 3 of the present application can regulate the metabolism of estrogen through the combination of Lactobacillus casei postbiotics and the medicinal and edible mixture, thereby fundamentally reducing the adverse effects of endocrine disorders on breast health.

[0141] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application. Various modifications and variations are possible for those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. A Lactobacillus casei postbiotic, characterized in that The Lactobacillus casei postbiotic is prepared by the following method: Inoculating Lactobacillus casei 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 Lactobacillus casei postbiotics; The Lactobacillus casei is Lactobacillus casei ( Lacticaseibacillus casei ) WZA-LC26, whose biological deposit number is CCTCC NO: M20241410; The fermentation medium is an MRS liquid medium as a base medium, and a medicine-food mixture having a mass fraction of 4-10% is added to the base medium; The medicinal and edible 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).

2. The Lactobacillus casei postbiotic according to claim 1, wherein The fermentation temperature is 30-37°C, and the fermentation time is 36-60h.

3. The Lactobacillus casei postbiotic according to claim 1, wherein The inoculation amount of Lactobacillus casei is 5%-15% of the volume of the fermentation medium.

4. Use of the Lactobacillus casei postbiotic according to any one of claims 1 to 3 in preparing a product for treating dysmenorrhea caused by cold coagulation and blood stasis.

5. Application of the Lactobacillus casei postbiotic according to any one of claims 1 to 3 in preparing a product for improving hyperplasia of the breast.

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