Application of Brawsonia in prevention and treatment of Parkinson's disease

Abstract

The invention provides an application of a Brawtella strain in regulation of intestinal flora and prevention and treatment of Parkinson's disease. The invention also provides application of the strain in preparation of probiotic preparations and preparation of drugs for preventing and treating Parkinson's disease, and the strain can change intestinal flora and influence the function of a central nervous system through an'intestine-brain axis' so as to treat and prevent Parkinson's disease.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and particularly relates to application of Blautia in preventing and treating Parkinson's disease. Background Art

[0002] Parkinson's disease (PD) is the second most common chronic progressive neurodegenerative disease after Alzheimer's disease. The disease is mainly manifested by symptoms such as tremor, muscle stiffness, bradykinesia and postural instability, which seriously affect the quality of life of patients. Epidemiological surveys show that with the aging of my country's population, it is estimated that by 2030, there will be about 5 million PD patients in my country, which will bring a heavy burden to society and the economy.

[0003] Currently, the treatment of Parkinson's disease mainly focuses on relieving symptoms, such as improving patients' motor function through dopamine replacement therapy. However, these treatments cannot delay or prevent the progression of the disease, and long-term use may cause serious side effects. Therefore, there is an urgent need to develop new treatments that can delay or prevent PD.

[0004] The pathogenesis of PD is still not fully understood. Recent studies have shown that the imbalance of intestinal flora is closely related to the occurrence of PD. As an important microecological system of the human body, the intestinal flora has complex interactions with the host's immune system, metabolic function and nervous system. More and more evidence shows that changes in the intestinal flora may affect the function of the central nervous system through the "gut-brain axis", thus playing an important role in the occurrence and development of PD.

[0005] By integrating human intestinal flora sequencing data, human blood mononuclear cell and human brain tissue transcriptome sequencing data, the researchers found that Blautia was the only genus that decreased in the feces, blood and brain of PD patients. Among them, the abundance of Blautia argi sp. (Blautia argi subspecies) decreased significantly in PD patients. This finding suggests that Blautia argi sp. may play an important role in the pathogenesis of PD. However, the specific mechanism of action of Blautiaargi sp. in PD is still unclear. Summary of the invention

[0006] The first objective of the present invention is to provide the use of the above-mentioned Blautia strain in regulating intestinal flora, preventing and treating Parkinson's disease. The strain can change the intestinal flora, affect the function of the central nervous system through the "gut-brain axis", and thus treat or prevent Parkinson's disease.

[0007] The second object of the present invention is to provide the use of the above-mentioned Blautia strain in the preparation of a probiotic preparation.

[0008] The third object of the present invention is to provide the use of the above-mentioned Blautia strain in the preparation of drugs for regulating intestinal flora.

[0009] The fourth object of the present invention is to provide the use of the above-mentioned Blautia strain in the preparation of a drug for preventing and / or treating Parkinson's disease.

[0010] The first technical solution adopted by the present invention is: application of Blautia strain in regulating intestinal flora and preventing and treating Parkinson's disease.

[0011] Furthermore, the Blautia strain regulates the intestinal flora by inhibiting the growth of harmful intestinal bacteria.

[0012] The second technical solution adopted by the present invention is: use of Blautia strains in the preparation of probiotic preparations.

[0013] Furthermore, probiotic preparations are used to regulate intestinal flora homeostasis and prevent and treat Parkinson's disease.

[0014] The third technical solution adopted by the present invention is: use of Blautia strains in the preparation of drugs for regulating intestinal flora.

[0015] The fourth technical solution adopted by the present invention is: use of Blautia strains in the preparation of medicines for preventing and treating Parkinson's disease.

[0016] Beneficial effects of the present invention:

[0017] The present invention has confirmed through clinical samples, in vivo and in vitro experiments that Blautia argi sp. can inhibit the growth of harmful intestinal bacteria, improve intestinal flora, have excellent resistance to Parkinson's disease and have no toxic side effects, can be used in a long-lasting and effective manner to prepare drugs for preventing and treating Parkinson's disease, and can be used to prevent and treat Parkinson's disease, with great application value. However, there is no prior art that Blautia argi sp. has the function of regulating intestinal flora and preventing and treating Parkinson's disease.

[0018] The present invention detects and finds that the content of Blautia argi bacteria in Parkinson's disease patients is significantly reduced, and the present invention finds that the abundance of Blautia argi bacteria in the intestinal flora is negatively correlated with the severity of Parkinson's disease; the Blautia argi strain of the present invention can inhibit the loss of dopaminergic neurons in the substantia nigra-striatum of PD rat models and improve the motor function of PD rats. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The abundance of Blautia argi sp. in the fecal flora of healthy controls and PD patients was detected;

[0020] Figure 2 is the relationship between the abundance of Blautia argi sp. and the severity of Parkinson's disease;

[0021] Figure 3 This is a flow chart of Blautia argi sp. intervention in animal experiments;

[0022] Figure 4 The effect of oral supplementation of intestinal flora Blautia argi sp. on motor function of PD rat model;

[0023] Figure 5 Oral supplementation of Blautia argi sp. in the intestinal flora can inhibit the loss of dopamine neurons in the substantia nigra of PD rat models;

[0024] Figure 6 Oral supplementation of Blautia argi sp. in the intestinal flora can inhibit the loss of dopamine neurons in the substantia nigra and striatum of PD rat models;

[0025] Figure 7 Oral supplementation of Blautia argi sp. can increase the abundance of Blautia argi in the intestinal flora and inhibit neuroinflammatory response in the brain of PD rat models. DETAILED DESCRIPTION

[0026] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0027] The present invention proposes a scheme for preventing and / or treating Parkinson's disease using Blautia argi sp. bacteria, and conducts relevant experimental studies. Preliminary results show that Blautia argi sp. bacteria has the potential to regulate intestinal flora and improve the function of the gut-brain axis, and thus may play a role in preventing and treating PD. The present invention aims to provide an innovative treatment method based on Blautia argi sp. bacteria, providing a new option for the treatment of Parkinson's disease.

[0028] The present invention provides the use of the above-mentioned Blautia strain in regulating intestinal flora and preventing and / or treating Parkinson's disease, and regulates intestinal flora by inhibiting the growth of harmful intestinal bacteria.

[0029] The present invention also provides the use of the above-mentioned Blautia strain in the preparation of a probiotic preparation, which is used to regulate the homeostasis of intestinal flora and prevent and treat Parkinson's disease.

[0030] The present invention also provides the use of the above-mentioned Blautia strain in preparing a drug for regulating intestinal flora.

[0031] The present invention also provides the use of the above-mentioned Blautia strain in preparing a medicine for preventing and / or treating Parkinson's disease.

[0032] The above-mentioned Blautia strain was purchased from the Korean Type Culture Collection (KCTC) by Ningbo Mingzhou Biotechnology Co., Ltd. The KCTC number is 15426 and the classification name is Blautia argi sp.

[0033] The present invention will be further described below by means of the accompanying drawings and embodiments.

[0034] Example 1

[0035] Abundance detection of Blautia bacteria in PD patients and healthy controls

[0036] Fecal samples of PD patients were provided by inpatients of the Department of Geriatric Neurology of Shaanxi Provincial People's Hospital. After collection, the samples were frozen in a -20℃ refrigerator in time and transported to the laboratory in a -80℃ refrigerator with dry ice within 2 days. Fecal samples of healthy people were provided by healthy people who underwent physical examinations and were taken back to the laboratory within 5 hours and frozen in a -80℃ refrigerator. When extracting fecal DNA, the samples were taken out from -80℃, and 100-150mg of feces were taken into a 1.5ml centrifuge tube with a medicine spoon without thawing; then, the total DNA of fecal flora was extracted using a fecal flora DNA extraction kit (Qiagen), and the DNA concentration, purity and quality were detected by Nanodrop 2000 UV microspectrophotometer and agarose gel electrophoresis. Subsequent operations were performed after passing the quality inspection. The samples were sent to Novogene for 16S rRNA sequencing to analyze the abundance of Blautia argi sp. in PD patients and normal people.

[0037] The results are as follows Figure 1 As described, the left side is a comparison diagram of the difference in abundance of the bacterial flora between PD patients and healthy controls; the right side is a comparison diagram of the difference in abundance of Blautia argi sp. between PD patients and healthy controls;

[0038] Depend on Figure 1It can be learned that the 16S rRNA gene sequencing technology was used to detect the distribution and relative abundance of intestinal flora in PD patients. The relative abundance of intestinal flora refers to the proportion of various microorganisms in the flora. The results of this study showed that compared with the control group, the abundance of the genus Blautia in the intestinal flora of PD patients decreased significantly (P=0.042). At the same time, it was found that the genera at the genus level of the intestinal flora of PD patients decreased compared with the control group, including Blautia, Phascolarctobacterium, Agarobacterium, Faecalibacterium, Bacteroides, and Parabacteroides, while the abundance of Bifidobacterium, Alternaria, Desulfovibrio, Escherichia-Shigella, and Acidococcus increased. Intestinal flora dysbiosis is closely related to the occurrence and development of PD, and the content of Blautia argisp. in the intestine of PD patients is significantly lower than that of healthy people.

[0039] Example 2

[0040] Association between the abundance of Blautia argi. in the intestinal flora and the severity of PD patients.

[0041] We analyzed the relationship between the abundance of Blautia in the intestinal flora of PD patients and PD-related indicators. The results of correlation analysis showed that the relative abundance of Blautia was negatively correlated with UPDRS-III (r=-0.680).

[0042] Depend on Figure 2 It can be seen that the UPDRS III score is correlated with the abundance of Blautia argi sp. bacteria: the content of Blautia bacteria in the intestinal flora is inversely proportional to the degree of PD development, that is, a high content of Blautia sp. bacteria can inhibit the development of PD, while a low content can aggravate the development of PD.

[0043] Example 3

[0044] The cylinder test is to observe the symmetry and usage rate of the forelimbs of experimental animals during the process of standing against the wall. Through the cylinder test, it was found that compared with the control group rats, the usage rate of the bilateral upper limbs of the PD rat model was significantly asymmetric, and the number of times the contralateral forelimbs touched the wall was significantly reduced (P=0.003), while the number of times the contralateral forelimbs touched the wall increased after oral administration of Blautia argi compared with the PD group (P=0.058). The body-raising and swinging test is to observe the flipping of experimental animals to both sides. We found that compared with the control group, the number of flipping to the contralateral side (left side) of the PD rat model after the body was raised was reduced (P=0.08), but Blautia argi failed to significantly improve the imbalance between the two sides. The open field test reflects the active ability of animals by recording the total distance of rat activities, average speed, number of times crossing the central grid, grooming, hind limb standing and other parameters. Our experimental results show that the number of upright standing times of rats in the experimental group is significantly reduced compared with the normal control group (P=0.001), and the number of upright standing times of rats is increased after Blautia argi intervention.

[0045] according to Figure 3 The intervention process in Figure 4 As shown, the left side of the first row shows the number of times the limb on the contralateral side of the injury hits the wall, the middle of the first row shows the number of times the limb flips to the healthy side, the right side of the first row shows the total distance, the left side of the second row shows the number of times the limb stands upright, the middle of the second row shows the number of times the limb crosses the central area, and the right side of the second row shows the number of times the limb grooms; **P<0.01, ***P<0.001,

[0046] It can be seen that after 10 weeks of intervention with Blautia argi bacteria transplantation, Blautia argi bacteria transplantation improved the bilateral motor coordination of the 6-OHDA-induced PD rat model, enhanced the movement and exploration ability of PD rats, and alleviated movement disorders.

[0047] Example 4

[0048] Blautia argi transplantation inhibits the loss of dopamine neurons in PD rat models and suppresses neuroinflammatory responses.

[0049] The typical characteristics of PD are motor symptoms and degeneration of dopamine neurons in the brain. To clarify the effect of Blautia argi on PD, we used immunohistochemistry to detect the number of dopamine neurons in the rat brain and counted them using ImageJ statistical software. TH is a key rate-limiting enzyme that mediates dopamine biosynthesis. The expression of TH is usually detected to reflect the severity of PD pathology. Its core principle is antigen-antibody reaction, that is, the detected antigen is bound by specific antibodies to clarify the pathological changes of dopamine neurons in the substantia nigra and striatum. Our results showed that 6-OHDA injection damaged TH-positive neurons in the ipsilateral substantia nigra of rats. After 11 weeks of injury, the number of TH-positive neurons in the ipsilateral substantia nigra was significantly reduced compared with the control group (P<0.001), and the TH area in the ipsilateral striatum was also significantly reduced compared with the control group (P<0.001). Early oral administration of Blautia argi to rats can significantly rescue the loss of dopamine neurons and their axons in the substantia nigra and striatum induced by 6-OHDA.

[0050] like Figure 5 As shown, the upper part is the staining result of substantia nigra dopamine neurons, and the lower part is the schematic diagram of the number of substantia nigra TH neurons and TH density. Compared with the control group: ***P<0.001; compared with the PD+Blautia group: ##P<0.01; Figure 6 As shown, the left side is the staining result of striatal dopamine neurons, and the right side is the schematic diagram of striatal TH density. Compared with the control group: ***P<0.001, compared with the PD+Blautia group: #P<0.05, ###P<0.001; Figure 7 As shown, the upper part is a schematic diagram of the difference in TH and GFAP in the substantia nigra (4X), and the lower part is a schematic diagram of the difference in TH and GFAP in the substantia nigra (20X). It can be seen that Blautiaargi bacteria transplantation inhibits the loss of dopamine neurons in the PD rat model and inhibits neuroinflammatory response.

Claims

1. Application of Blautia strains in regulating intestinal flora and preventing and treating Parkinson's disease.

2. Use of the Blautia strain according to claim 1 in regulating intestinal flora and preventing and treating Parkinson's disease, wherein the Blautia strain regulates intestinal flora by inhibiting the growth of harmful intestinal bacteria.

3. Application of Blautia strains in the preparation of probiotic preparations.

4. The use of the Blautia strain according to claim 3 in the preparation of a probiotic preparation, characterized in that: The probiotic preparation is used for regulating the homeostasis of intestinal flora and preventing and treating Parkinson's disease.

5. Application of Blautia strains in the preparation of drugs for regulating intestinal flora.

6. Use of Blautia strains in the preparation of drugs for preventing and treating Parkinson's disease.

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