Application of heat shock protein gp96 in treatment of rheumatoid arthritis

A heat shock protein and rheumatoid technology, applied in the field of biomedicine, can solve the problem that rheumatoid arthritis cannot be cured completely.

Active Publication Date: 2016-10-26
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, rheumatoid arthritis cannot be cured no matter whether it is treated by traditional Chinese medicine or western medicine. It can only be relieved by drugs, which brings huge economic and mental burdens to RA patients and their families.

Method used

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  • Application of heat shock protein gp96 in treatment of rheumatoid arthritis
  • Application of heat shock protein gp96 in treatment of rheumatoid arthritis
  • Application of heat shock protein gp96 in treatment of rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1, the extraction of pgp96

[0096] The extraction steps of heat shock protein gp96 (hereinafter referred to as pgp96) in tissues are as follows:

[0097] (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.

[0098] (2) After completing step (1), centrifuge at 16500 g for 1 h to obtain supernatant A.

[0099] (3) After completing step (2), take the supernatant A and centrifuge at 16500 g for 50 min to obtain the supernatant B.

[0100] (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.

[0101] (5) After completing step (4), load the sample solution onto the ConA Sepharose column.

[0102] (6) After step (5) is completed,...

Embodiment 2

[0108] Embodiment 2, the preparation of recombinant heat shock protein gp96

[0109] 1. Recombinant plasmid pFastBac TM 1- Construction of gp96

[0110] 1. The RNA of HepG2 cells was extracted by Trizol method, and then cDNA was obtained by reverse transcription.

[0111] 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).

[0112] 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.

[0113] 4. Double-digest the PCR amplification product with restriction endonucleases EcoRI and XbaI, and recover the digested product.

[0114] 5. Digest plasmid ...

Embodiment 3

[0131] Example 3, Determination of the dosage of pgp96 or rgp96 in activated regulatory T cells

[0132] 1. Immunization of mice in groups

[0133] 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:

[0134] 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.

[0135] pgp96 treatme...

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Abstract

The invention discloses application of a heat shock protein gp96 in the treatment of rheumatoid arthritis. The heat shock protein gp96 is (a1) or (a2) or (a3) or (a4): (a1) a protein with an amino acid sequence represented from a site 2 to a site 783 from a terminal N in a sequence 2 in a sequence table; (a2) a protein with an amino acid sequence represented by the sequence 2 in the sequence table; (a3) a fusion protein obtained by linking a label to the terminal N or / and a terminal C of (a1) or (a2); and (a4) a protein, with the same functions, obtained through substitution and / or deletion and / or addition of one or more amino acid residue from the site 2 to the site 783 from the terminal N in the sequence 2 in the sequence table or the amino acid sequence represented by the sequence 2 in the sequence table. Experiments prove that the heat shock protein gp96 has important application values in the treatment of rheumatoid arthritis and / or the relief of symptoms of the rheumatoid arthritis.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of heat shock protein gp96 in treating rheumatoid arthritis. Background technique [0002] Rheumatoid arthritis (rheumatoid arthritis, RA) is a chronic, inflammatory, systemic autoimmune disease, characterized by non-suppurative inflammation of joints and joint tissues, mainly manifested as synovitis of the joints, eventually leading to Damage to various tissues such as cartilage, ligaments, tendons of the joints, and multiple organs. The age of onset of 80% of RA patients is between 20 and 45 years old, and the incidence rate of the population is 1% to 2%. Among them, women are three times that of men. The basic pathological changes of RA are synovitis, synovial swelling, exudation, and granulocyte infiltration in the acute phase; hyperplasia and hypertrophy of the synovial membrane in the chronic phase, and the formation of vascular gaps, which cause join...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P19/02A61P29/00
CPCA61K38/1709
Inventor 孟颂东郑华国
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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