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Lactobacillus casei with immunomodulatory, anti-inflammatory and anti-cervical cancer effects and application

A technology of Lactobacillus casei and immune regulation, applied in the field of microorganisms, can solve problems such as different mechanisms, achieve the effect of preventing weight loss, wide application prospects, and improving colitis symptoms

Active Publication Date: 2020-08-21
TIANJIN UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] In conclusion, although Lactobacillus casei has been reported in the published patents or patent applications for the relief of enteritis, the mechanisms are different. It can regulate immunity, regulate the balance of M1 / M2 macrophages, and increase Treg cells. The number of Lactobacillus casei has not yet been retrieved, and the Lactobacillus casei that can inhibit the expression of E6 and E7 has an anti-cervical cancer effect has not been reported

Method used

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  • Lactobacillus casei with immunomodulatory, anti-inflammatory and anti-cervical cancer effects and application
  • Lactobacillus casei with immunomodulatory, anti-inflammatory and anti-cervical cancer effects and application
  • Lactobacillus casei with immunomodulatory, anti-inflammatory and anti-cervical cancer effects and application

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Experimental program
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Effect test

Embodiment 1

[0060] Embodiment 1: Screening and identification of strains

[0061] (1) Isolation and purification of strains

[0062] Grind the koji evenly with a sterile mortar, weigh 1g of koji into 10mL of normal saline, and dilute to 10 -6 , take 10 -4 、10 -5 、10 -6 Spread 100 μl of the three gradient dilutions on the MRS solid plate, incubate at 37°C for 48 hours, select colonies of different shapes and separate them by streaking, until a pure single colony with the same colony shape is obtained, pick a single colony and inoculate it in 5ml of MRS liquid In the culture medium, cultivate overnight, take 800 μl of bacterial liquid and add 200 μl of 60% glycerol to store at -80°C.

[0063] (2) Identification of strains

[0064] The isolated strains were subjected to PCR amplification of 16S rDNA, and the PCR products were sent to Jinweizhi for sequencing. The sequencing results were compared in NCBI to obtain a strain of Lactobacillus casei, which was named Lactobacillus casei LH23....

Embodiment 2

[0067] Embodiment 2: Lactobacillus casei LH23 acid resistance determination

[0068] The lactic acid bacteria were activated for 3 times, placed in an anaerobic incubator at 37°C, and cultured overnight; the concentration of the bacteria was diluted to 5×10 8 CFU / mL, replace the MRS medium with different pH values, the pH is 2.0, 3.0, 4.0 respectively; put the bacterial solution with different pH in a 96-well plate, set 6 parallel wells for each experimental group, and set up a negative control , and its negative control is MRS with a pH value of 7, which is placed in an anaerobic incubator at 37°C for 4 hours. 20 μL of MTT (5 mg / mL) will be added and placed in an anaerobic incubator for 4 hours. Measure the absorbance value of each hole at 490nm (A experimental group) with an enzyme-linked immunosorbent detector, and compare it with the absorbance value of the negative control group (A control group) to determine the survival rate. The survival rate calculation formula is a...

Embodiment 3

[0071] Embodiment 3: Lactobacillus casei LH23 bile salt resistance determination

[0072] The lactic acid bacteria were activated for 3 times, placed in an anaerobic incubator at 37°C, and cultured overnight; the concentration of the bacteria was diluted to 5×10 8 CFU / mL, in the MRS medium that replaces different bile salt concentrations, the concentration of bile salt is 0.1% (w / v), 0.2% (w / v), 0.3% (w / v); method to measure survival.

[0073] The result is as figure 2 As shown, Pediococcus acidillactici ML2523, Lactobacillus caseiLH23, L.acidop hilus ML4523, Lactobacillus casei LH2723, these 4 strains of lactic acid bacteria were mixed with 0.1% (w / v), 0.2% (w / v), 0.3% (w / v) respectively The difference in survival rate after 4h cultured in MRS liquid medium with bile salts was significant. Among them, the ability of LH23 to tolerate bile salts at 0.1%-0.3% was significantly stronger than other strains. When bile salts were 0.1%, LH23 was 118.92%±2.19, and when bile salts ...

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Abstract

The invention relates to lactobacillus casei with immunoregulation, anti-inflammatory and anti-cervical cancer effects. The lactobacillus casei strain is named as LH23, the preservation number is CGMCC No.16656, and the strain can be used for regulating immunity, regulating M1 / M2 macrophage balance, increasing the number of Treg cells, improving the symptoms of mouse colitis, shortening the colonlength and apoptosis of intestinal epithelial cells, inhibiting excessive secretion of proinflammatory factors (TNF-alpha, IL-1beta and IL-6) and recovering the concentration of short-chain fatty acid(SCFAs) in the intestinal tract, can resist cervical cancer, inhibit proliferation of cervical cancer HeLa cells and acetylation modification of histone, cause apoptosis of the HeLa cells and inhibitexpression of human papilloma virus HPVE6 and E7 proteins. The strain is used for preparing fermented food for inhibiting inflammatory bowel diseases and cervical cancer, and has a very wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of microbes, in particular to a Lactobacillus casei with immune regulation, anti-inflammation and anti-cervical cancer effects and its application. Background technique [0002] Inflammatory bowel disease (IBD) is a group of gastrointestinal inflammatory diseases with unclear pathogenesis, mainly including ulcerative colitis (UC) and Crohn's disease (CD) ). Neither of these two gastrointestinal inflammations is fatal, but they affect people's normal life, manifested as abdominal pain, diarrhea, nutritional disorders, and blood in the stool. According to the analysis of the literature on IBD epidemiology in the past 50 years, the incidence of IBD is the highest in Europe at present, but the incidence of IBD is gradually increasing with the westernization of lifestyle in many developing countries. [0003] The main drug treatments in modern medicine mainly include aminoglycosides, glucocorticoids, immunosupp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/747A61P37/02A61P29/00A61P35/00A23C9/123A23L11/00A23L19/00A23L33/135C12R1/245
CPCA61K35/747A61P37/02A61P29/00A61P35/00A23C9/1234A23L19/00A23L33/135A23V2002/00A23L11/50C12R2001/245C12N1/205A23V2400/125A23V2200/324A23V2200/308A23V2200/32
Inventor 王楠何红鹏张同存刘美玲郝云鹏申敬张秀霞扈士月凌涛罗学刚齐威李中媛宋亚囝
Owner TIANJIN UNIV OF SCI & TECH
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