Preparation method and application of nano material with synovitis inhibition function

A technology of nanomaterials and inflammation, applied in the direction of nanotechnology, nanotechnology, nanomedicine, etc., can solve problems such as pain, loss of function, limitations, etc., and achieve the effect of alleviating the inflammatory cascade reaction

Pending Publication Date: 2021-12-17
WEST CHINA HOSPITAL SICHUAN UNIV
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Problems solved by technology

[0005] At present, OA is the most common joint disease in the elderly population in the world. OA has become an important cause of the life quality of the elderly

Method used

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  • Preparation method and application of nano material with synovitis inhibition function
  • Preparation method and application of nano material with synovitis inhibition function
  • Preparation method and application of nano material with synovitis inhibition function

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Embodiment 1

[0035] Embodiment 1: the preparation method of the nanomaterial with synovial inflammation inhibition function, comprises the following steps:

[0036] S1. Construction of shRNA-LeptinR: Insert the synthetic shRNA-LeptinR fragment into the Plko.1-EGFP-PURO plasmid vector to obtain a recombinant plasmid vector, then transform the recombinant plasmid vector into Escherichia coli, and extract to obtain shRNA-LeptinR plasmid, the shRNA-LeptinR plasmid contains a green fluorescent protein marker fragment, and has ampicillin resistance, and then the shRNA-LeptinR plasmid is transformed into Escherichia coli, and the forward sequence of the primer used is 5' -CACAAAGAAGTTACATCTT-3'; the reverse sequence of the constructed primer is 3'-AAGATGTAACTTCTTTGTG-5'.

[0037] S2. Macrophage membrane preparation:

[0038] A. Take the mouse mononuclear macrophage leukemia cells (M0) in the logarithmic growth phase, and when the density reaches 70%, add 20ng·mL -1 After IL-4 and IL-13 acted fo...

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Abstract

The invention discloses a preparation method and application of a nano material with a synovitis inhibition function, and belongs to the technical field of biology. According to the key point of the technical scheme, the preparation method comprises the following steps that S1, shRNA-LeptinR is constructed, specifically, a synthesized shRNA-LeptinR fragment is inserted into a plasmid vector to obtain a recombinant plasmid vector, then, the recombinant plasmid vector is transformed into escherichia coli, and the shRNA-LeptinR plasmid is obtained through extraction; S2, a macrophage membrane is prepared; and S3, gene-loaded nanoparticles M2H@RPK is prepared, specifically, the nanoparticles M2H@RPK is prepared by adopting an electrostatic adsorption method, the nanoparticles H@RPK is wrapped with the M2 macrophage membrane extracted in the step S2, and a polycarbonate membrane repeatedly passes through by using a micro extruder to obtain the nanoparticles M2H@RPK. The preparation method is mainly used for relieving the inflammatory cascade reaction in the OA process, and the nano-system targets the cartilage matrix and can effectively relieve joint injury and inhibit the severity of overall arthritis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method and application of a nanometer material with the function of inhibiting synovium inflammation. Background technique [0002] Accumulating evidence indicates that osteoarthritis (OA) is an inflammation-related immune disease in which the synovial membrane plays an important role. The synovial membrane of the joint can be divided into two layers, namely the inner synovial layer and the subsynovial layer which are thinner and close to the joint cavity. Clearly demarcated, they are macrophage-like synoviocytes and fibroblast-like synoviocytes. [0003] During synovitis, the number of macrophages increases dramatically. This massive increase is partly due to the escape of the apoptotic pathway by the original macrophages in the synovium, but more from the infiltrating macrophages in the blood circulation. Macrophages are highly variable cells that respond to differ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P19/02A61P19/04A61P29/00B82Y5/00B82Y40/00
CPCA61K48/0033A61K48/005A61P19/04A61P19/02A61P29/00B82Y5/00B82Y40/00Y02A50/30
Inventor 杨成莉周凯胡丹蓉钱志勇
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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