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Application of heat shock protein gp96 to treatment of systemic lupus erythematosus

A heat shock protein and lupus erythematosus technology, applied in the field of biomedicine, can solve the problems that SLE cannot be cured

Active Publication Date: 2016-09-28
FOSHAN HEAT SHOCK BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, no matter whether SLE is treated with traditional Chinese medicine or Western medicine, it is impossible to cure it. It can only be relieved by drugs, which brings huge economic and mental burdens to SLE patients and their families.

Method used

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  • Application of heat shock protein gp96 to treatment of systemic lupus erythematosus
  • Application of heat shock protein gp96 to treatment of systemic lupus erythematosus
  • Application of heat shock protein gp96 to treatment of systemic lupus erythematosus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example 1, the extraction of pgp96

[0097] The extraction steps of heat shock protein gp96 (hereinafter referred to as pgp96) in tissues are as follows:

[0098] (1) Isolate the placental tissue of the mouse before delivery to obtain the isolated mouse placental tissue. Take isolated mouse placental tissue or human isolated placental tissue, cut it into pieces, add solution A according to the mass volume ratio of 1g: 4mL, and then grind it with a glass homogenizer.

[0099] (2) After completing step (1), centrifuge at 16500 g for 1 h to obtain supernatant A.

[0100] (3) After completing step (2), take the supernatant A and centrifuge at 16500 g for 50 min to obtain the supernatant B.

[0101] (4) After completing step (3), take the supernatant B, add solution B at a volume ratio of 9:1, and mix well to obtain the sample solution.

[0102] (5) After completing step (4), load the sample solution onto the ConA Sepharose column.

[0103] (6) After step (5) is completed,...

Embodiment 2

[0109] Embodiment 2, the preparation of recombinant heat shock protein gp96

[0110] 1. Recombinant plasmid pFastBac TM 1- Construction of gp96

[0111] 1. The RNA of HepG2 cells was extracted by Trizol method, and then cDNA was obtained by reverse transcription.

[0112] 2. According to the sequence of human gp96 gene (GenBank No. AY040226.1), artificially synthesized primer F1: 5'-G GAATTC ATGGACGATGAAGTTGAT-3' (the underline is the restriction endonuclease EcoRI recognition sequence) and R1: 5'-GC TCTAGA CTATTAGAATTCATCTTTTTC-3' (the underline is the recognition sequence of restriction endonuclease XbaI).

[0113] 3. After completing steps 1 and 2, use the cDNA obtained in step 1 as a template and F1 and R1 synthesized in step 2 as primers to perform PCR amplification to obtain PCR amplification products.

[0114] 4. Double-digest the PCR amplification product with restriction endonucleases EcoRI and XbaI, and recover the digested product.

[0115] 5. Digest plasmid ...

Embodiment 3

[0132] Example 3, Determination of the dosage of pgp96 or rgp96 in activated regulatory T cells

[0133] 1. Immunization of mice in groups

[0134] Ninety nine-week-old mice with a body weight of 18-22 g were randomly divided into pgp96 treatment group 1, pgp96 treatment group 2, pgp96 treatment group 3, pgp96 treatment group 4, rgp96 treatment group 1, rgp96 treatment group 2, rgp96 treatment group 3 , rgp96 treatment group 4 and control group (every group of 10), carry out the following treatments respectively:

[0135] pgp96 treatment group 1: on the first day of the experiment, the solution of pgp96 prepared in Example 1 was injected intraperitoneally; on the eighth day of the experiment, the solution of pgp96 prepared in Example 1 was injected intraperitoneally again; on the 22nd day of the experiment, the solution prepared in Example 1 was injected again intraperitoneally A solution of pgp96. The dose of each injection was 10 μg pgp96 per mouse.

[0136] pgp96 treatme...

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Abstract

The invention discloses an application of a heat shock protein gp96 to treatment of systemic lupus erythematosus. The heat shock protein gp96 is a1) or a2) or a3) or a4): the a1) amino acid sequence represents a protein including a second amino acid site from a N terminal to a 783th amino acid site shown in a sequence 2 in a sequence table; the a2) amino acid sequence represents a protein shown in the sequence 2 in the sequence table; the a3) amino acid sequence represents a fused protein obtained by connecting a label to a N terminal or / and a C terminal of the a1) or the a2); the a4) amino acid sequence represents a protein with the same function which is obtained after replacement and / or deletion and / or addition of one or several amino acid residues is carried out for the protein including the second amino acid site from the N terminal to the 783th amino acid site shown in a sequence 2 or the amino acid sequence shown in the sequence 2 in the sequence table. Experiments prove that the heat shock protein gp96 has important application values for treatment of systemic lupus erythematosus and / or alleviation of symptoms of systemic lupus erythematosus.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to the application of heat shock protein gp96 in treating systemic lupus erythematosus. Background technique [0002] Systemic lupus erythematosus (SLE) is an autoimmune disease that usually occurs in women aged 20-40. A large number of studies have shown that genetic, endocrine, infection, immune abnormalities and some environmental factors are related to the pathogenesis of SLE, but the etiology has not yet been determined. Under the interaction of genetic factors, environmental factors, estrogen levels and other factors, SLE patients have decreased T lymphocytes, decreased T suppressor cell function, excessive proliferation of B cells, and produced a large number of autoantibodies. These autoantibodies can combine with the corresponding autoantigens in the body to form corresponding immune complexes, which are deposited on the skin, joints, small blood vessels, glomeruli, e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61P37/02A61P19/04
Inventor 孟颂东李鑫陈密
Owner FOSHAN HEAT SHOCK BIOTECH CO LTD
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