Bifidobacterium longum for relieving insulin resistance and application thereof
A technology of Bifidobacterium longum and Bifidobacterium, applied in the field of microbiology, can solve problems such as heavy economic burden for patients, difficulty in maintaining blood sugar homeostasis, and adverse reactions in the digestive tract, so as to reduce liver triglyceride and inflammation levels, and relieve liver inflammation. Tissue damage, reducing the effect of insulin resistance index
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[0062] Preparation of activated medium (g / L): Components include carbon source: pectin 0.047, xylan 0.047, arabinogalactan, pullulan 0.04, soluble starch 0.392; nitrogen source: bactopeptone 24, tryptone 24. Inorganic salts: magnesium sulfate heptahydrate 0.5, potassium dihydrogen phosphate 2.5, sodium chloride 4.5, calcium chloride dihydrate 0.45, iron sulfate heptahydrate 0.005; bile salt 0.4, anaerobic cysteine hydrochloride 0.2 and acid-base buffer (MES) 19.52. Firstly, the above ingredients are prepared, and the pH is adjusted to 6, followed by deoxygenation and sterilization (121° C., 15 min). After the sterilization, the medium was transferred to an anaerobic glove box, and 1 μg of high-temperature-labile heme, 1 μg of vitamin K3 (VK3) and 0.1 mL of vitamin mixture solution (Wolfe’s Vitamin Solution) were added to 1 L of culture medium through a 0.22 μm filter membrane. The base was deaerated overnight in an anaerobic glove box to obtain the activated liquid medium. ...
Embodiment 1
[0086] Example 1: Isolation and screening of Bifidobacterium longum NSP008
[0087] 1. Sample collection
[0088] Collect human feces samples of type Ⅱ diabetes in Shangqiu, Henan Province, place the samples in preservation tubes, add 5 times the weight of protective solution (preparation of protective agent: weigh cysteine hydrochloride 1g / L, glycerol 200-300g / L, uniformly dissolved in PBS (1×), sterilized at 115-121°C for 15-20min), stored in an insulated box with dry ice, brought back to the laboratory, and immediately placed in a -80°C refrigerator to be separated and screened.
[0089] 2. Enrichment of fecal bacteria
[0090] Take the above-mentioned fecal bacteria liquid out of the -80°C refrigerator, and after thawing, centrifuge at low speed and low temperature (500g, 5min, 4°C) to obtain the supernatant, then pass through a 100μm filter membrane to remove impurities in the supernatant, and inoculate the supernatant fecal bacteria liquid into the activated culture ...
Embodiment 2
[0099] Example 2: Effects of Bifidobacterium longum NSP008 on body weight, body fat and diet of insulin-resistant mice on a high-fat diet
[0100] Specific steps are as follows:
[0101] 1. Preparation of Bifidobacterium longum NSP008 cryopreservation agent:
[0102] (1) Culture method: In a sterile anaerobic environment, the Bifidobacterium longum NSP008 bacterial classification was streaked on the MRS solid medium, cultivated under anaerobic conditions for 48 hours, after forming a single colony, pick a single colony, and It was inoculated into MRS liquid medium, and cultured anaerobically at 37° C. for 16-24 hours to reach a stationary phase, at which time the OD value was 1.0-1.4, and a seed solution was prepared.
[0103] (2) Preparation of protective agent: Weigh 1 g / L of cysteine hydrochloride and 200-300 g / L of glycerol, dissolve them evenly in distilled water, and sterilize at 115-121° C. for 15-20 min.
[0104] (3) Preparation of refrigerant: Centrifuge (8000rpm,...
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