Application of blautia sp B2132 bacteria in prevention and/or treatment of inflammatory bowel disease
A technology for inflammatory bowel disease and Crohn's disease, applied in the field of microorganisms, can solve the problems of low abundance of Lachnospiraceae and lack of Lachnospiraceae strains, etc., achieve significant application value, improve intestinal flora, Effect without toxic side effects
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Embodiment 1
[0047] Example 1, Isolation, cultivation and identification of Blautia sp.B2132 bacteria
[0048] (1) Conditioned medium preparation
[0049] Pre-deoxygenated liquid BHI medium (containing 5% bovine serum and 0.1% cysteine): Weigh 37 g of brain heart infusion (BD company) into a 1 L Erlenmeyer flask, add 1 g of cysteine and double distilled Dilute the water to 1000ml, stir to dissolve, and then sterilize in an autoclave at 121°C for 15 minutes; after the sterilization is completed, put it into an anaerobic incubator, cool it to about 40-50°C, add 50ml of fetal bovine serum and Shake evenly to obtain pre-deoxygenated liquid BHI medium, and then divide into 50ml centrifuge tubes.
[0050] Pre-deoxygenated solid BHI medium (containing 5% bovine serum, 0.1% cysteine and 1 mg / L aztreonam): take 37 g of brain heart infusion (BD company) in a conical flask with a capacity of 1 L, add 1g cysteine, 15g agar powder and double-distilled water to 1000ml, stir to dissolve, and then s...
Embodiment 2
[0067] Example 2, Abundance Detection of Blautia Bacteria Content in IBD Patients and Healthy Controls
[0068] (1) Fecal flora DNA extraction
[0069] Stool samples from IBD patients were provided by inpatients in the Inflammatory Bowel Disease Center of Run Run Shaw Hospital Affiliated to Zhejiang University. After collection, the samples were frozen and stored in a -20°C refrigerator in time, and transported to the laboratory with dry ice in a -80°C refrigerator within 2 days. The fecal samples of healthy people were provided by healthy people for physical examination, and they were taken back to the laboratory within 5 hours and stored in a -80°C refrigerator. When extracting fecal DNA, take the sample from -80°C, and take 100-150 mg of feces with a medicine spoon into a 1.5ml centrifuge tube without thawing; then, use the fecal flora DNA extraction kit (Qiagen) to extract the total DNA of the fecal flora , and then use a Nanodrop 2000 UV micro-spectrophotometer and agaro...
Embodiment 3
[0080] Embodiment 3, Blautia sp. Bacterial abundance in the intestinal flora has a great impact on Il-10 - / - Effects of spontaneous enteritis in mice.
[0081] Il-10 knockout mice (Il-10 - / - The immune cells of mice) cannot produce interleukin 10 (Interleukin 10, IL-10), so under normal feeding conditions, the IL-10 of about 12 weeks old - / - Mice develop spontaneous enteritis, especially colonic inflammation, characterized by an inflammatory infiltration of lymphocytes, macrophages, and neutrophils. The severity of the disease is closely related to the structural composition of the intestinal flora. Therefore the present invention utilizes Il-10 - / - A mouse model of spontaneous enteritis to evaluate the relationship between Blautia sp. and the pathogenesis of enteritis.
[0082] First, sex, age matched 10 Il-10 - / - Mice were housed in an SPF environment for up to 8 weeks. Then, use the quantitative PCR method to detect 10 Il-10 - / - The relative abundance of Blautia sp. ...
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