Lactobacillus fermentum HY01 with effects of intestinal tract function regulation and colitis prevention, and uses thereof

A technology of Lactobacillus fermentum and intestinal function, applied in the field of microorganisms, can solve the problems of different mechanisms of action, side effects, and high prices

Inactive Publication Date: 2017-09-15
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these drugs can prevent colitis to varying degrees, their mechanisms of action are different, and most of them have the characteristics of side effects and high price, so they are not suitable for ordinary patients
For example, long-term use of 5-aminosalicylic acid can cause abdominal pain, nausea, hemolytic anemia, and renal failure; colitis relapses are higher with low doses of glucocorticoids, while high doses can cause cataracts and osteoporosis In addition, the use of antibiotics can change the living environment of intestinal flora, which is not conducive to the survival of beneficial microorganisms
In addition, many researchers also pay attention to the effect of traditional Chinese medicine preparations on the treatment of colitis, such as Huanglian decoction, Wumei pill, Shenling Baizhu powder, Gegen Qinlian decoction, Sibai decoction and Sishen pill, etc. Inflammation prevention effect, but traditional Chinese medicine preparations have the characteristics of complex composition and unknown ingredients

Method used

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  • Lactobacillus fermentum HY01 with effects of intestinal tract function regulation and colitis prevention, and uses thereof
  • Lactobacillus fermentum HY01 with effects of intestinal tract function regulation and colitis prevention, and uses thereof
  • Lactobacillus fermentum HY01 with effects of intestinal tract function regulation and colitis prevention, and uses thereof

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Experimental program
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Effect test

Embodiment 1

[0061] The isolation, purification and preliminary identification of Lactobacillus fermentum (Lactobacillus fermentum) HY01, this embodiment is carried out according to the following steps:

[0062] (1) Isolation and purification of Lactobacillus fermentum HY01

[0063] 1 mL of the sample (naturally fermented yak yoghurt from herdsmen's home in Hongyuan County, Sichuan Province) was drawn by aseptic operation and added to 9 mL of sterilized saline to make a uniform dilution of 1:10, and continue to make a certain proportion of dilution. Select a suitable gradient dilution solution, pipette 100 μL each into the MRS solid plate medium with a sterile pipette tip, and incubate at 37°C for 48 hours, observe and record the colony morphology, such as figure 1 shown. Use an inoculation loop to pick different colonies from the surface of the plate and inoculate them in MRS liquid medium, culture them on a shaker at 37°C and 100r / min for 18 hours; figure 2 As shown, it is determined ...

Embodiment 2

[0068] In Vitro Screening of Lactobacillus fermentum HY01

[0069] Lactic acid bacteria are part of the normal intestinal flora. As one of the first conditions for probiotics, they can survive in the stomach and small intestine. The strains to be screened should have sufficient acid resistance and growth and development characteristics in acidic environments. It is necessary for fermentation The acid resistance of Lactobacillus fermentum HY01 was analyzed.

[0070] (1) Screening of probiotics resistant to pH 3.0 artificial gastric juice

[0071] Preparation of artificial gastric juice: 0.2% NaCl, 0.35% pepsin, adjust the pH value to 3.0 with 1M HCl, filter and sterilize with a vacuum pump in an aseptic operating table for later use.

[0072] Determination of the tolerance of probiotics to artificial gastric juice: take 5mL of the activated strain culture solution, pour it into a sterilized 10mL centrifuge tube in a sterile operating table, collect the bacteria by centrifugati...

Embodiment 3

[0081] DSS (dextran sulfate sodium)-induced colitis experiment: 40 Kunming mice were randomly divided into 4 groups, 10 in each group, half male and half female, and the experiment started after one week of feeding, and the experiment period was five weeks. The feeding conditions of mice in each group were as follows: during the experiment, they were kept in stainless steel cages, the room temperature was kept at 25±2°C, the relative humidity was 50±10%, and light and dark were alternated for 12 hours. The mice in each group were free to eat. In addition, mice in the normal group were free to drink water (without DSS) every day until the end of the experiment. The mice in the model group had free access to water (without DSS) in the first, second and fourth weeks, water containing 2% DSS in the third week, and water containing 4% DSS in the fifth week. Low concentration group (10 9 CFU / kg·BW) and high concentration group (10 10 CFU / kg·BW) During the experiment, Lactobacillus...

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Abstract

The present invention discloses Lactobacillus fermentum HY01 with effects of intestinal tract function regulation and colitis prevention, and uses thereof, wherein the preservation number of the Lactobacillus fermentum HY01 is CCTCCM2015792. According to the present invention, the in vitro tolerance of the Lactobacillus fermentum HY01 is strong, wherein the survival rate of the Lactobacillus fermentum HY01 is 103.73+ / -8.60% after the Lactobacillus fermentum HY01 is treated for 3 h in the artificial gastric juice with the pH value of 3.0, the Lactobacillus fermentum HY01 can grow at the bile salt concentration of 0.3%, the growth efficiency is 21.62+ / -0.86% of the growth efficiency of the bile salt-free culture, and the Lactobacillus fermentum HY01 has good in vitro tolerance; and the in vivo mouse test results show that the Lactobacillus fermentum HY01 can alleviate the colonic shortening and the colonic edema of the colitis mice, and the inflammatory cell infiltration and the mucosal injury in colon, such that the Lactobacillus fermentum HY01 has the colitis prevention effect.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Lactobacillus fermentum HY01 capable of regulating intestinal function and preventing colitis and its use. Background technique [0002] Inflammatory bowel disease is a chronic non-specific intestinal disease whose etiology and pathogenesis are unclear. Colitis is one of them. Great pain and distress. Colitis occurs worldwide and is related to race, region, and age. There is no significant difference between men and women in the incidence population. According to statistics, the incidence rate of colitis in western countries is the highest, the incidence rate is 1.2-20.3 / 100,000, and the prevalence rate is 7.6-245 / 100,000; the incidence rate in Europe is 19.2-24.3 / 100,000, and the incidence rate in Asia and the Middle East The rate is 6.3 / 100,000. However, due to the improvement of people's living standards and the change of diet structure in our country, the incidenc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23C9/127A23L33/135C12R1/225
CPCA23C9/127A23V2002/00A23L33/135C12N1/205C12R2001/225A23V2200/3204
Inventor 索化夷曾涛李键赵欣骞宇陈孝勇
Owner SOUTHWEST UNIV
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