Application of lactobacillus muricatum in preparation of medicine for preventing and / or treating Alzheimer disease and medicine containing lactobacillus muricatum

Abstract

The invention discloses application of lactobacillus muricatum in preparation of a medicine for preventing and / or treating Alzheimer's disease in the technical field of microorganisms, and the lactobacillus muricatum is preserved in the China Industrial Microbiological Culture Collection Center (CICC) with the preservation number of CICC23140. Research finds that the inactivated lactobacillus muricatum can effectively protect hippocampal neurons and improve cognitive impairment, and the effect of preventing and treating the Alzheimer's disease is good.

Description

Technical Field

[0001] The present invention belongs to the technical field of microorganisms, and particularly relates to an application of Lactobacillus murinus in preparing a drug for preventing and / or treating Alzheimer's disease and a drug containing the Lactobacillus murinus. Background Art

[0002] Alzheimer's disease (AD) is a degenerative disease of the central nervous system that occurs mainly in the elderly. The main clinical manifestations are progressive decline in cognitive memory function and daily living ability, accompanied by a variety of neuropsychiatric symptoms and behavioral disorders. It is the most widespread dementia disease with the largest number of patients in the world. The main pathological characteristics of AD include a large loss of cholinergic neurons and their synapses in the cerebral cortex and hippocampus, vascular amyloid degeneration of cortical arteries and arterioles, accumulation of amyloid β (Aβ), and abnormal hyperphosphorylation of tau protein to form neurofibrillary tangles. The exact cause of this pathological change is still unclear. Studies have shown that genetic factors, environmental factors, aging, neurotransmitter system dysfunction, inflammatory response, neurotoxic damage, oxidative stress, etc. are all related to the occurrence of AD.

[0003] In recent years, researchers have proposed that microorganisms from the intestine (including pathogenic microorganisms) play an important role in the development of AD pathology. Experiments have shown that the intestine may be a key source of Aβ in the brain, and the intestinal microbiota can upregulate intestinal Aβ, which affects the occurrence and development of AD after entering the brain. The microbiota secretes a variety of metabolites involved in human growth, development and pathological processes, including short chain fatty acids (SCFAs), bile acids and neuroactive molecules. SCFA is the most abundant metabolite and comes from indigestible dietary fiber metabolized by intestinal microorganisms. Studies have shown that the content of short-chain fatty acids SCFAs and bacteria that produce SCFAs in the intestine of AD patients is significantly lower than that of normal controls. In AD mouse models, sodium butyrate can improve cognitive impairment in the early and late stages of AD, which confirms that there is a certain relationship between SCFAs deficiency and AD. Abnormal changes in SCFAs concentrations have been observed in AD. They participate in the AD process by regulating synaptic plasticity, Aβ and tau pathology, and neuroinflammation. The pathogenesis of AD has not yet been identified, which has brought great difficulties to the treatment of AD. The symptoms of AD gradually worsen as the disease progresses. Unfortunately, there is no medical treatment to eradicate Parkinson's disease. Clinical treatment of Parkinson's disease is mostly based on Aβ 25-35 However, drug resistance caused by long-term administration and the side effects of the drug itself have greatly limited the treatment of the disease. Therefore, determining the cause of AD and finding an effective treatment plan are still urgent issues that need to be solved.

[0004] Lactobacillus murinus is a Gram-positive bacterium belonging to the family Lactobacillus in the phylum Firmicutes. It is mainly isolated from the intestines of mice. According to literature research, Lactobacillus murinus can repair damaged mucosa and enhance the intestinal barrier. It also has the function of regulating intestinal microbiota. However, there are no reports on the effect of Lactobacillus murinus on Alzheimer's disease. Summary of the invention

[0005] The present invention aims to provide an application of Lactobacillus murinus in the preparation of a drug for preventing and / or treating Alzheimer's disease and a drug containing the Lactobacillus murinus. The Lactobacillus murinus provided by the present invention can protect hippocampal neurons, improve the cognitive function of AD rats, reduce the expression of inflammatory factors COX2, IL-1β and IL-18 proteins in the hippocampus of rats, and has a good effect in preventing and treating Alzheimer's disease.

[0006] In the first aspect, the present invention provides a use of Lactobacillus murinus in the preparation of a drug for preventing and / or treating Alzheimer's disease, wherein the Lactobacillus murinus is deposited in the China Industrial Microbiological Culture Collection Center with a deposit number of CICC23140. The Alzheimer's disease is preferably Aβ 25-35 Induced Alzheimer's disease.

[0007] Furthermore, the Lactobacillus ratus is an inactivated Lactobacillus ratus.

[0008] In a second aspect, the Lactobacillus murine described in the first aspect can also be used to prepare drugs for improving cognitive impairment and / or saving hippocampal neuron loss and / or reducing the level of inflammatory factors in peripheral blood.

[0009] Furthermore, the inflammatory factors in the peripheral blood include any one or more of IL-6, TNF-α and IL-1β.

[0010] In a third aspect, the present invention further provides a drug, the composition of which includes Lactobacillus murinus as described in the first aspect above, and has the effects of the first and second aspects above.

[0011] Further, the concentration of inactivated Lactobacillus murinus in the drug is 10 6 ~10 12 cfu / ml, preferably 10 9 cfu / ml.

[0012] Furthermore, the composition of the drug also includes one or more of starch, maltodextrin, trehalose and lactose.

[0013] Furthermore, the dosage form of the drug is selected to be any one of a suspension, suppository, tablet, capsule, soft capsule, prefilled injection, effervescent tablet or gel.

[0014] The beneficial technical effect of the present invention is that the present invention has found that the inactivated Lactobacillus murinus, which is deposited in the China Industrial Microbiological Culture Collection Center and has a deposit number of CICC23140, is particularly effective in improving and / or preventing and / or treating Aβ. 25-35 The invention has a good effect on the induced Alzheimer's disease, can effectively protect hippocampal neurons, reduce the expression of inflammatory factors in the brain, and improve the cognitive impairment of Alzheimer's disease. In addition, the Lactobacillus murinus has many advantages such as excellent effect, high safety, good stability, mature production process, and wide source of raw materials, so it is suitable for promotion and use in clinical practice. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 It is a bar chart showing the number of times each group of rats crossed the platform during the platform detection period of the water maze experiment;

[0016] Figure 2 This is a representative trajectory diagram of the rat water maze experiment;

[0017] Figure 3 This is a Nissl staining image of the rat hippocampus and cortex;

[0018] Figure 4 It is the NEUN immunofluorescence image and mean fluorescence intensity bar graph of rat hippocampus and cortex;

[0019] Figure 5 It is a bar chart of ELISA testing of inflammatory factors IL-6, TNF-α and IL-1β in peripheral blood;

[0020] Figure 6 It is the expression bands and bar charts of inflammatory factors COX2, IL-1β and IL-18 proteins in the hippocampus;

[0021] Meaning of the reference numerals in the drawings of the specification:

[0022] Control group, sham operation group;

[0023] HK Lm indicates sham operation + Lactobacillus murinus group;

[0024] Aβ 25-35 represents the model group;

[0025] Aβ 25-35 +HK Lm indicates Aβ 25-35 Model+Lactobacillus murinus group. DETAILED DESCRIPTION

[0026] The following is further described in detail through specific implementation methods:

[0027] The present invention has found that Lactobacillus murinus, which is deposited in the China Industrial Microbiological Culture Collection Center and has a deposit number of CICC23140, can be used to prepare a method for improving and / or preventing and / or treating Aβ. 25-35 Drugs that induce Alzheimer's disease and improve Aβ 25-35 It can induce cognitive impairment and hippocampal neuron damage in AD rats, reduce the levels of inflammatory factors IL-6, TNF-α and IL-1β proteins in the peripheral blood, and at the same time reduce the expression of inflammatory factors COX-2, IL-1β and IL-18 proteins in the hippocampus, and has a good preventive and therapeutic effect on AD.

[0028] The drug uses the inactivated Lactobacillus murinus as the main active ingredient and also includes pharmaceutically acceptable excipients, which can play a role in improving patient compliance and improving the stability of the inactivated Lactobacillus murinus. The excipients can be selected from one or more of starch, maltodextrin, trehalose and lactose, preferably trehalose. The dosage form of the drug includes suspension, suppository, tablet, capsule, soft capsule, prefilled injection, effervescent tablet and gel. The present invention has no special requirements for the preparation method of the above dosage form, and the preparation method well known in the art can be used.

[0029] In the present invention, the preparation method of the inactivated Lactobacillus murinus preferably comprises the following steps:

[0030] The Lactobacillus murine is sterilized to obtain an inactivated Lactobacillus murine.

[0031] The present invention has no particular limitation on the sterilization method, and conventional sterilization methods may be used, such as inactivation at 121° C. for 20 to 30 minutes, more preferably inactivation at 121° C. for 20 minutes.

[0032] In the present invention, the concentration of the inactivated Lactobacillus murinus in the drug is preferably 10 6 ~10 12 CFU / ml is more preferably 10 9 cfu / ml. The inactivated Lactobacillus murinus of the above concentration can better play the role of preventing and treating Alzheimer's disease.

[0033] In the present invention, Aβ 25-35 Taking the induced Alzheimer's disease rat model as an example, the technical effect of the inactivated Lactobacillus murineus in improving and treating Alzheimer's disease is further introduced in detail.

[0034] experiment:

[0035] 1.1 Experimental animals

[0036] SD male rats, weighing about 200 g, were purchased from the Experimental Animal Center of Zunyi Medical University and housed in the SPF animal room of the Key Laboratory of Basic Pharmacology of the Ministry of Education of Zunyi Medical University. Each group of experimental animals was kept under constant temperature and humidity conditions, with free access to clean drinking water and basic feed, and a circadian rhythm of 12 h.

[0037] 1.2 Main instruments and equipment

[0038] Rat water maze, rat open field, rat Y maze, rat elevated plus maze, Bio-Rad chemiluminescence imaging system (Bio-Rad, USA), BSS-110 electronic analytical balance (Beijing Sartorius Electronic Balance Co., Ltd.), Milli QA pure water processor (Millipore Corporation).

[0039] 1.3 Main experimental reagents

[0040] Lactobacillus murinus (Lactobacillus murinus, China Industrial Microbiological Culture Collection Management Center, MRS medium (casein peptone 10.0g, beef extract 10.0g, yeast extract powder 4.0g, glucose 20.0g, sodium acetate 5.0g, triammonium citrate 2.0g, Tween 80 1.0ml, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, distilled water 1000 mL), pH 6.2 (Solarbio), Aβ 25-35 (purity ≥ 98%, Sigma), neuronal nucleus (NeuN) rabbit polyclonal antibody (Proteintech), microtubule-associated protein 2 (MAP2) rabbit polyclonal antibody (Proteintech), Nissl staining solution (toluidine blue method).

[0041] 2. Experimental Methods

[0042] 2.1 Grouping and Dosing

[0043] SD male rats, weighing about 200 g, were randomly divided into 4 groups, 10 rats in each group. 25-35 Group, Aβ 25-35 +HK Lm group.

[0044] 1) Sham operation group: After 4 weeks of drinking water treatment, normal saline was injected into the brain stereotaxically, and then the same volume of drinking water was continuously given for 3 weeks.

[0045] 2) HK Lm group: heat-inactivated Lactobacillus murinus 10 9 After 4 weeks of pretreatment with cfu / day, the brain was stereotaxically injected with saline, followed by heat-inactivated Lactobacillus murinus for 10 9 cfu / day for 3 weeks.

[0046] 3) Aβ 25-35 Group: After 4 weeks of drinking water treatment, 3u L of 5ug / ul Aβ was injected stereotactically 25-35 Solution was given to establish an Alzheimer's disease model, and then drinking water was continuously administered for 3 weeks.

[0047] 4) Aβ 25-35 +HK Lm group: heat-inactivated Lactobacillus murinus 10 9 After 4 weeks of pretreatment with cfu / day, 3uL of Aβ at a concentration of 5ug / uL was injected stereotaxically 25-35 Solution was used to establish the Alzheimer's disease model, and then heat-killed Lactobacillus murinus was continuously administered for 10 9 cfu / day for 3 weeks.

[0048] 2.2 AD animal model preparation

[0049] The rats were deeply anesthetized by intraperitoneal injection of 3% sodium pentobarbital. The hair on the top of the rats' heads was removed and the rats were fixed on a rat brain stereotaxic instrument. The shaved area was disinfected with iodine tincture, and the rat's scalp was cut to expose the anterior fontanelle. With the anterior fontanelle as the origin of the coordinates, 3uL Aβ with a concentration of 5ug / uL was uniformly injected at 3.6mm behind the anterior fontanelle, 2.0mm to the left of the midline, and 3.6mm below the dura mater. 25-35 Solution, stop the injection for 5 minutes after the injection, then take out the syringe at a constant speed, suture the wound, apply erythromycin ointment, and continue the experiment.

[0050] 2.3 Lactobacillus murinus culture and administration

[0051] Lactobacillus murine culture: First, activate the bacteria in the freeze-dried tube and perform subculture to restore the activity of Lactobacillus murine. Inoculate Lactobacillus murine in MRS medium and culture at 37°C for 8 h. Measure the OD value of the bacterial solution with an ultraviolet spectrophotometer at 600 nm. The OD value is about 0.8, and the total number of bacteria is 2.8×10 8cfu / mL, centrifuge the suspension, discard the supernatant and adjust the bacterial solution concentration to 10 with purified water 8 ~10 9 cfu / mL to obtain Live Lm, and then inactivate at 121℃ for 20min to obtain Hk Lm. The total amount of bacteria administered by gavage was 1×10 9 cfu / mL.

[0052] 2.4 Behavioral testing

[0053] 2.4.1 Morris water maze test

[0054] Three weeks after modeling, the rats began to undergo behavioral water maze testing. The experiment was divided into a positioning navigation experiment and a space exploration experiment. Positioning navigation experiment: The rats received training for 5 consecutive days, 4 times a day, with an interval of 30 minutes between each training. The time required for the rats to enter the water from the 4 water entry points and find the platform was recorded, i.e., the escape latency. The average score of the 4 latency periods was used as the final result of the day and entered the final statistics. Space exploration experiment: On the 6th day of the experiment, the platform was removed, and the rats were placed in the water after entering the water from the farthest end of the platform. The swimming trajectory of the rats within 120 seconds was recorded, and the number of times the rats crossed the platform in the target quadrant was observed and analyzed.

[0055] 2.5 Sampling and specimen preparation

[0056] After completing the behavioral experiment, the rats were deeply anesthetized, the abdominal cavity was cut open, the abdominal aorta was clamped, the chest cavity was cut open, the heart was fully exposed, the perfusion needle was inserted into the aorta through the apex of the heart, the right atrial appendage was fixed with hemostatic forceps and cut open, and PBS was perfused until the blood in the brain was washed clean. The whole brains of 5 rats in each group were removed, and the hippocampus and cortex were separated and stored at -80℃ respectively.

[0057] 2.6 Nissl staining

[0058] Used to observe changes in brain neurons. Paraffin sections (5 μm) were baked in a 60°C oven for 1 hour, dewaxed by conventional xylene-ethanol, washed with double distilled water for 5 minutes × 3 times, 100 μL of toluidine blue solution was taken to cover the tissue, stained in a 60°C incubator for 40 minutes, washed with double distilled water for 5 minutes × 3 times; rapidly differentiated in 95% ethanol for 2 seconds, dehydrated with anhydrous ethanol for 2 seconds, air-dried in a fume hood, and sealed with neutral gum. Observed under an upright microscope.

[0059] 2.7 Immunofluorescence

[0060] 1) Dewaxing of paraffin sections: sequentially place the sections in xylene I for 15 min, xylene II for 15 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, 85% alcohol for 5 min, 75% alcohol for 5 min, and wash with distilled water.

[0061] 2) Antigen repair: Place the tissue sections in a repair box filled with EDTA antigen repair buffer (PH 8.0) in a microwave oven for antigen repair. Heat on medium heat for 6 min × 3 times. During this process, prevent the buffer from evaporating excessively. Fill up the repair solution after each time. After natural cooling, place the slides in PBS (PH 7.4) and wash them on a decolorizing shaker for 3 times, 5 min each time.

[0062] 3) Draw circles: After the slices are slightly dried, use a histochemical pen to draw circles around the tissue (to prevent the antibodies from flowing away).

[0063] 4) Serum blocking: Add 10% goat serum in the circle and incubate for 40 minutes.

[0064] 5) Add primary antibody: Gently shake off the blocking solution, add 1:300 NEUN primary antibody prepared with PBS on the slices, and lay the slices flat in a humidified box and incubate at 4°C overnight.

[0065] 6) Add secondary antibody: Place the slide in PBS (PH7.4) and wash on a decolorizing shaker for 3 times, 5 minutes each time. After the slices are slightly dried, add rabbit-derived green light secondary antibody at a dilution of 1:800 to cover the tissue and incubate at room temperature for 50 minutes.

[0066] 7) DAPI counterstaining of cell nuclei: After the slices are slightly dried, DAPI staining solution is added to the circle and incubated at room temperature in the dark for 10 minutes.

[0067] 8) Autofluorescence quenching: After the slices are slightly dried, add autofluorescence quencher in the circle for 5 minutes and rinse with running water for 10 minutes.

[0068] 9) Sealing: Place the slide in PBS (PH7.4) and wash on a decolorizing shaker for 3 times, 5 minutes each time. After the slices are slightly dried, seal them with anti-fluorescence quenching sealing medium.

[0069] 2.8ELISA method to detect the activity of inflammatory factors IL-6, IL-1β and TNF-α in rat serum

[0070] Take rat serum and dispense about 230 μL per tube. After equilibrating the ELISA kit at room temperature for 10 minutes, dilute the standard, add 100 μL of protein sample and standard to each well, incubate at 37°C for 2 hours, dry and add 100 μL of detection solution A working solution, incubate at 37°C for 1 hour, discard the liquid in the well and repeat washing 3 times, add 100 μL of detection solution B working solution to each well, incubate at 37°C for 30 minutes, repeat washing 5 times, add 90 μL of substrate solution to each well, color at 37°C in the dark, add stop solution and detect the absorbance value of each well at a wavelength of 450 nm, calculate the concentration value of the corresponding protein according to the standard curve, and the result is expressed in ng / μg protein.

[0071] 2.9 Western blotting

[0072] 1) Weigh an appropriate amount of hippocampal tissue, grind it on ice, add RIPA lysis buffer (containing PMSF), and let it stand for 40 minutes;

[0073] 2) Centrifuge in an ultra-low temperature centrifuge at 12000 rpm for 10 min and collect the supernatant;

[0074] 3) Quantify using the BCA protein quantification kit, prepare the sample loading system, mix well, and denature at 100°C for 10 minutes;

[0075] 4) Add 10 μL to each well of 10% precast gel and perform electrophoresis for 2 h (voltage 80-110 V);

[0076] 5) Use wet transfer method to transfer the membrane. Place the filter paper, polyvinylidene fluoride membrane (PVDF membrane), gel, and filter paper in the order of electroporation box, set the current to 300 mA, and transfer for 40 minutes;

[0077] 6) After electroporation, block with 5% skim milk for 2 h and wash with TBST 3 times; 10 min / time

[0078] 6) Add primary antibody and incubate at 4°C overnight, then wash with TBST 3 times, 10 min / time;

[0079] 7) Add secondary antibody and incubate at 4°C for 40 min, then wash with TBST three times, 10 min each time;

[0080] 8) Add exposure agent and expose.

[0081] 2.8 Statistical analysis of data

[0082] The experimental data were statistically analyzed using Graphpad prism statistical software. All data were expressed as (mean ± standard deviation). One-way analysis of variance (ANOVA) was used to compare group differences in means. The LSD method was used to test for homogeneity of variance. Dunnett's T3 was used to compare group differences in means. P < 0.05 was considered statistically significant.

[0083] 3. Experimental results

[0084] 3.1 Effect of inactivated Lactobacillus murinus on memory in rats with Alzheimer's disease

[0085] Behavioral experiments were performed 21 days after AD modeling. Figure 1 As shown, after 21 days of AD modeling, Aβ 25-35 The number of times the rats in the group crossed the original platform position decreased, which was related to Aβ 25-35 Compared with the Aβ 25-35The number of times the rats in the +HK Lm group crossed the original platform position increased significantly, and was statistically significant (p<0.05). This showed that the AD model was successfully established, and it can be seen that the early administration of inactivated Lactobacillus murinus can improve Aβ 25-35 induced memory ability in AD rats.

[0086] 3.2 Changes of Nissl bodies in the hippocampal DG region and cortex

[0087] like Figure 3 As shown, compared with the Control group, Aβ 25-35 The neurons in the DG and cortex of the group were disordered, accompanied by cell edema and necrosis, and the Nissl bodies were stained lightly and unclearly. 25-35 Compared with the Aβ 25-35 In the +HK Lm group, the Nissl bodies of neurons in the DG region and cortex were arranged more regularly, and edema and dissolution of neurons were occasionally observed. There was a statistically significant difference (p<0.05).

[0088] 3.3 Immunofluorescence staining of NEUN

[0089] like Figure 4 As shown, compared with the Control group, Aβ 25-35 The NEUN fluorescence intensity in the DG and cortex of the group was reduced, which was consistent with Aβ 25-35 Compared with the Aβ 25-35 In the +HK Lm group, the NEUN fluorescence intensity in the DG area and cortex was significantly increased, and the difference was statistically significant (p<0.05).

[0090] 3.5ELISA to measure the expression of inflammatory factors IL-6, IL-1β and TNF-α in rat serum

[0091] Depend on Figure 5 As shown, inactivated Lactobacillus murinus can reduce Aβ 25-35 Expression of IL-6, IL-1β and TNF-α protein levels in the serum of rats with induced Alzheimer's disease.

[0092] 3.6 Effect of inactivated Lactobacillus murinus on Aβ 25-35 Effects of inflammatory factors COX2, IL-1β and IL-18 on the rat Alzheimer's disease model induced by

[0093] Depend on Figure 6 As shown, compared with Control, Aβ 25-35 The expression levels of COX2, IL-1β and IL-18 proteins in the group A were significantly decreased, and the difference was statistically significant (P<0.05). 25-35 Compared with the Aβ 25-35The expression levels of COX2, IL-1β and IL-18 proteins in the +HK Lm group were significantly increased, and the differences were statistically significant (P<0.05).

[0094] Summary: From the perspective of overall experiment, the present invention verifies that inactivated Lactobacillus murinus has the following beneficial effects through methods such as water maze, etc., thereby proving that inactivated Lactobacillus murinus has a therapeutic effect on Alzheimer's disease and can delay its occurrence and development.

[0095] The invention is beneficial in that: the invention provides an application of inactivated Lactobacillus murinus, that is, inactivated Lactobacillus murinus in the preparation of a drug for preventing or treating Aβ 25-35 Application of drugs that induce Alzheimer's disease. 25-35 Induces Alzheimer's disease and has a preventive and therapeutic effect.

[0096] (1) Lactobacillus murinus can improve cognitive impairment in rats to a certain extent and can improve Aβ 25-35 Induced Alzheimer's disease.

[0097] (2) Effect of Lactobacillus murinus on Aβ 25-35 It plays a protective role in the hippocampal neuron damage induced by Alzheimer's disease in rats.

[0098] (3) Lactobacillus murinus can reduce Aβ 25-35 Expression levels of inflammatory factors in hippocampal tissue of rats with induced Alzheimer's disease.

[0099] In summary, Lactobacillus murinus can improve cognitive impairment in Alzheimer's disease and has a positive effect on Aβ 25-35 It can delay or improve the induced Alzheimer's disease and is suitable for promotion and use in clinical practice.

Claims

1. A use of Lactobacillus murinus in the preparation of a drug for preventing and / or treating Alzheimer's disease, characterized in that: The Lactobacillus murinus is deposited in the China Industrial Microbiological Culture Collection Center with a deposit number of CICC23140.

2. The use according to claim 1, characterized in that: The Alzheimer's disease is preferably Aβ 25-35 Induced Alzheimer's disease.

3. Use of Lactobacillus murinus in the preparation of a drug for improving cognitive impairment or / and saving hippocampal neuron loss or / and reducing the level of inflammatory factors in peripheral blood, characterized in that: The Lactobacillus murinus is deposited in the China Industrial Microbiological Culture Collection Center with a deposit number of CICC23140.

4. The use according to claim 3, characterized in that: The inflammatory factors in the peripheral blood include any one or more of IL-6, TNF-α and IL-1β.

5. The use according to any one of claims 1 to 4, characterized in that: The Lactobacillus ratus is inactivated Lactobacillus ratus.

6. A drug, characterized in that The composition of the medicine includes the Lactobacillus murine as described in claim 5.

7. A medicine according to claim 6, characterized in that: The concentration of inactivated Lactobacillus murinus in the drug is 10 6 ~10 12 cfu / ml.

8. A medicine according to claim 7, characterized in that: The concentration of the inactivated Lactobacillus murinus in the drug is preferably 10 9 cfu / ml.

9. A medicine according to claim 6, characterized in that: The composition of the medicine also includes one or more of starch, maltodextrin, trehalose and lactose.

10. A medicine according to claim 6, characterized in that: The dosage form of the drug is selected from any one of suspension, suppository, tablet, capsule, soft capsule, prefilled injection, effervescent tablet or gel.

Images