Preparation method of manlam and its application in the preparation of medicines for treating autoimmune diseases

A technology for autoimmune diseases and drugs, applied in the field of biomedicine to achieve the effect of alleviating IBD disease and reducing the effect of the disease

Active Publication Date: 2019-04-12
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no report on the use of ManLAM extracted and purified from Mycobacterium tuberculosis in the treatment of autoimmune diseases

Method used

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  • Preparation method of manlam and its application in the preparation of medicines for treating autoimmune diseases
  • Preparation method of manlam and its application in the preparation of medicines for treating autoimmune diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] [Example 1] Preparation of ManLAM

[0045] After extracting Mycobacterium tuberculosis ManLAM, the crude sugar was first identified by glycogen staining, and its main components were still two kinds of polysaccharides. between kD ( figure 1 A) The isolated and purified ManLAM on the surface of Mycobacterium tuberculosis was lyophilized and then dissolved in PBS to adjust the concentration to 1 mg / mL. Before scanning, PBS was used to adjust the scanning baseline, using a wavelength scanner from 900 nm to 190 nm at intervals of 1 nm Continuous wavelength scanning showed that ManLAM had the highest absorption peak at 202 nm; then the purity was identified by Agilent 1260 HPLC, and its retention time was measured at 6.3 min, the main peak appeared, and its content was calculated according to the peak area to account for about 95% ( figure 1 B); after purification of ManLAM, the results of Western blot detection using Biotin-ConA showed a concentrated mannoglycoside-rich b...

Embodiment 2

[0091] [Example 2] ManLAM-stimulated immune cells adoptively relieve symptoms of inflammatory bowel disease

[0092] In dextran sodium sulfate (dextran sodium sulfate, DSS)-induced inflammatory bowel disease (inflammatory bowel disease, IBD), IL-10 secreted by B cells can inhibit neutrophil infiltration and activate CD4+ T cells And the secretion of IFN-γ, etc., thereby significantly slowing down the symptoms of the disease. To further explore whether ManLAM-induced B10 cells can exert immunosuppressive effects in inflammatory bowel disease models, we adopted ManLAM-treated fluorescent dye CFSE-labeled B cells into IL-10 - / - In mice, then observe its regulatory effect on the progression of inflammatory bowel disease.

[0093] After adopting ManLAM-treated B cells for 3 days, IL-10- / - mice were fed with purified water containing DSS (3%, w / v) to induce an acute inflammatory bowel disease model. Detection of body weight changes found: adoption of WT B cells and IL-10 without a...

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Abstract

The invention discloses a preparation method of ManLAM and application of ManLAM to preparation of a medicine for treating autoimmune diseases. The preparation method comprises the following steps: obtaining a raw sugar extract from mycobacterium tuberculosis, dissolving with a PBS so as to obtain a raw sugar solute, performing SDS-PAGE, putting a whole piece of separation gel into a 3% precooledpotassium chloride solution, and shaking slightly till the separation gel has milk white strips; cutting strips near 34 kD; putting the cut strips into a sterile mortar, repeatedly grinding, and soaking the grinded gel strips into deionized water of two times of the volume; centrifuging 10,000 g of the strips for 15 minutes, collecting supernate, and performing freeze drying, thereby obtaining ManLAM of which the purity is not less than 95%. Immune cells IL-10 stimulated by ManLAM are increased, an immunity regulation function is achieved, immune cell Th1 type polarization of bodies can be degraded, and the substance, namely ManLAM, has a treatment effect on all autoimmune diseases caused by Th1 type polarization increase.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for extracting and purifying ManLAM from mycobacterium conjunctiva and its application in the preparation of medicines for treating autoimmune diseases. Background technique [0002] Autoimmune diseases (ADs) are chronic and complex diseases that seriously endanger human health. They are caused by immune effector cells such as cytotoxic T lymphocytes (CTL), natural killer cells (NK cells), etc., or a class of chronic, heterogeneous diseases in which immune effector molecules (complement, antibodies, cytokines, etc.) produce pathological immune responses against self-tissues or cells, which may involve specific target organs or multiple systems. Including more than 20 kinds of diseases such as organ-specific autoimmune diseases (such as insulin-dependent diabetes mellitus) and systemic autoimmune diseases (such as systemic lupus erythematosus and rheumatoi...

Claims

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Application Information

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Patent Type & AuthorityPatents(China)
IPC IPC(8): C08B37/00A61K31/715A61P37/02A61P1/00A61P19/02A61P29/00
CPCA61K31/715C08B37/00
Inventor章晓联
OwnerWUHAN UNIV