Bifidobacterium breve capable of down-regulating IL-17 and relieving constipation and application of bifidobacterium breve
A technology of bifidobacteria breve and microbial inoculum, applied in the field of microorganisms, can solve the problems of single function of bifidobacteria and some inconspicuous functions, and achieve the effect of improving the water content of feces and maintaining good vitality
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Embodiment 1
[0065] Example 1: Acquisition of Bifidobacterium breve (B. breve) CCFM1260
[0066] Specific steps are as follows:
[0067] 1. Isolation and screening of Bifidobacterium strains:
[0068] (1) The feces of 9-month-old males in Beijing were collected using a disposable sterile urinal, and the fecal samples were placed in a liquid medium containing MRS+mass percent (0.05%-0.1%) cysteine containing fructooligosaccharides, and the Anaerobic incubator (N 2 :CO 2 :H 2 =80:10:10) for 12h enrichment;
[0069] (2) The fecal sample was diluted with sterile physiological saline and then applied to MRS+ mass percentage (0.05%-0.1%) L- On the solid plate of cysteine hydrochloride, cultivate for 24-48h;
[0070] (3) choose the single colony that meets the basic form of bifidobacteria to carry out streak purification on the plate, and screen and isolate the selected strain of bifidobacteria;
[0071] (4) The above single colony was cultured in liquid MRS+mass percentage (0.05%-0.1%...
Embodiment 2
[0090] Example 2: Bifidobacterium breve (B. breve) CCFM1260 relieves constipation-related symptoms induced by loperamide
[0091] Specific steps are as follows:
[0092] (1) Preparation of Bifidobacterium breve CCFM1260
[0093] Bifidobacterium breve CCFM1260 strains were taken out from the -80°C refrigerator, streaked in MRS solid medium, cultured anaerobic at 37°C for 48h, and a single colony was picked in MRS liquid medium, anaerobic culture at 37°C for 24h, prepared seed solution;
[0094] The prepared seed liquid was inoculated into a new MRS liquid medium with an inoculum of 2% (v / v), cultured anaerobic at 37°C for 24h, and cultured for another generation in the same way to prepare Bifidobacterium breve CCFM1260 fermentation broth;
[0095] Then the prepared Bifidobacterium breve CCFM1260 fermentation broth was centrifuged at 6000 r / min and 4°C for 5 min, and resuspended with 10% skim milk to obtain a bacterial suspension, which could be used in animal experiments.
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Embodiment 3
[0107] Example 3: Bifidobacterium breve (B. breve) CCFM1260 down-regulates the content of interleukin 17 (IL-17) in colon tissue of constipated mice
[0108] (1) The methods of grouping, modeling and treatment of BABL / c mice were the same as those in Example 2.
[0109] (2) After the mice were sacrificed on the 36th day, the collected mouse colon tissue was crushed and ground to make a homogenate, centrifuged at 12,000 g for 15 min to take the supernatant, and the mouse interleukin 17 detection kit was used, and the procedure was carried out according to the instructions. In the experiment, the concentration of interleukin 17 in colon tissue was calculated from the standard curve, and the total protein content of colon tissue was measured by BCA total protein kit, and the final result was the ratio of interleukin 17 to total protein concentration.
[0110] The experimental results are as figure 2 shown, by figure 2 It can be seen that after loperamide was used for modeling...
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