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Microanaerobic bacteria and application thereof for treating colonic inflammation

A microanaerobic and bacterial technology, applied in the fields of medicine and microbiology, can solve the problem of treating colitis and other problems that have not yet been discovered by Helicobacter hepatica

Active Publication Date: 2020-06-23
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are no research reports and patents on the use of Helicobacter hepaticus (H.hepaticus) to treat colitis and prevent colon cancer.
Previous studies on Hepatic Helicobacter strains mostly focused on the strain ATCC 51449, and all believed that ATCC 51449 bacteria is a pathogenic bacteria that can cause chronic hepatitis and colitis

Method used

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  • Microanaerobic bacteria and application thereof for treating colonic inflammation
  • Microanaerobic bacteria and application thereof for treating colonic inflammation
  • Microanaerobic bacteria and application thereof for treating colonic inflammation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The cultivation of embodiment 1 Helicobacter hepatica 51448

[0034] 1.1 Preparation of Helicobacter hepatica 51448 liquid medium (1L)

[0035] Prepared with BD's brussels broth. Add 28g of brussels broth solid powder per liter of double-distilled water in the Erlenmeyer flask, help dissolve at room temperature on a magnetic stirrer until completely dissolved, and carry out high-pressure sterilization at 121° C. for 20 minutes. When the culture medium drops to about 55°C, add 5% premium fetal bovine serum, mix and pour on the plate, which is the liquid medium for cultivating 51448. An appropriate amount of antibiotics can also be added to the culture medium to inhibit the growth of miscellaneous bacteria. For example: vancomycin: final concentration 10 μg / mL, polymyxin: final concentration 0.38 μg / mL, amphotericin: final concentration 2 μg / mL.

[0036] 1.2 Preparation of Helicobacter hepatica 51448 agar plate (1L)

[0037] Prepared with BD's Brucella agar. Add 43g ...

Embodiment 2

[0052] Example 2 Sequencing verification of Helicobacter hepatica 51448

[0053] 2.1 Test items

[0054] Detection material: the 51448 bacterial liquid samples cultivated in Example 1;

[0055] Detection: detect the bacterial species information of the sample.

[0056] 2.2 Detection method

[0057] Genomic DNA of strains was extracted by rapid genome extraction method for PCR amplification.

[0058] PCR amplification fungal 16S rDNA region, the primer sequence is as follows

[0059] F: AgAgTTTgATCCTggCTCAg (SEQ ID NO: 1)

[0060] R: CTACggCTACCTTgTTACgA (SEQ ID NO: 2)

[0061] 2.3 Test results

[0062] The length of the amplified product was about 1500bp, and the sequence information of the 16S rDNA region was obtained. Aligned by the online sequence comparison software BLAST (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi), the matching degree with ATCC5144816S rRNA, Sequence ID: NR_114584.1, is over 99%. The result is as Figure 8 It shows that Helicobacter hepatica 51448...

Embodiment 3

[0063] Example 3 Effect of Helicobacter hepatica 51448 on colon cancer

[0064] 3.1 Construction of colon cancer mouse model

[0065] We randomly divided C57 / B6 male mice aged 6-8 weeks into two groups, 12 in each group, which were normal saline control group and Helicobacter hepaticus 51448 experimental group. First, Azoxymethane (AOM) and dextran sulfate (Dextran Sulfate Sodium, DSS) were used to jointly induce the two groups of animal models to establish a mouse model of colorectal cancer, 5 days in advance (denoted as time point -5 days) ) according to the dose of 12.5mg / kg to mice intraperitoneally injected with AOM. After 5 days (recorded as time point 0 day), drink three-distilled water containing 2% DSS for one week (recorded as time point 7 days), and then change to drink normal water for two weeks (recorded as time point 21 days), according to every 21 days as a cycle, a total of 4 cycles, a total of 84 days, and then the mice were fed under normal drinking conditi...

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Abstract

The invention discloses a microanaerobic bacterium for treating colon inflammation, and the microanaerobic bacterium is Helicobacter hepatica. The present invention also provides the application of the microanaerobic bacteria in the preparation of drugs for preventing and / or treating colon inflammation and the application in the preparation of drugs for preventing colon cancer. In the present invention, the inventors clearly proposed for the first time that Helicobacter helicobacter can treat inflammatory enteritis and prevent colon cancer. It provides important clinical value and reference significance for the intestinal flora to diagnose and treat colorectal cancer.

Description

technical field [0001] The invention relates to the fields of microbiology and medical technology, in particular to microanaerobic bacteria for treating colon inflammation and applications thereof. Background technique [0002] Colorectal cancer (CRC) is one of the most common malignant tumors of the digestive tract. Statistics show that the incidence of colon cancer ranks fourth in males and third in females. However, at this stage there has been a lack of effective methods and drugs for the treatment and prevention of colorectal cancer. Recent studies have shown that giving patients probiotics to maintain the homeostasis of their intestinal flora is one of the effective means to improve intestinal inflammation and prevent colon cancer. However, at this stage, research on the types, optimal concentrations, treatment methods and courses of intestinal probiotics is still relatively lacking. [0003] Helicobacter hepaticus (H. hepaticus) is a Gram-negative bacillus, which is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/741A61P1/00A61P35/00C12R1/01
CPCA61K2035/11C12N1/20C12N1/205C12R2001/01
Inventor 张纪岩王晶花蕾赵敏吴会芳黄晓峰偏丽丽沈倍奋
Owner ACADEMY OF MILITARY MEDICAL SCI
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