Research method of protection mechanism of BMSC-Exo on high glucose induced HK2 cell damage

A technology of cell damage and protection mechanism, applied in biochemical equipment and methods, cells modified by introducing foreign genetic material, pharmaceutical formulations, etc., can solve the problem that MSCs cannot explain the therapeutic effect, and achieve the prevention of rapid degradation and high biocompatibility. Capacitance, economical and simple effect of process preparation

Inactive Publication Date: 2020-12-22
SOUTHWEST MEDICAL UNIVERISTY
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Problems solved by technology

However, studies in recent years have found that the number of initially implanted MSCs that survived, differentiated, and fused into injured tissues in situ is far from explaining the observed therapeutic effects.

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  • Research method of protection mechanism of BMSC-Exo on high glucose induced HK2 cell damage
  • Research method of protection mechanism of BMSC-Exo on high glucose induced HK2 cell damage
  • Research method of protection mechanism of BMSC-Exo on high glucose induced HK2 cell damage

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Embodiment

[0074] see Figure 1-6 , the present invention provides a technical solution: BMSC-Exo to the research method of high glucose-induced HK2 cell damage protection mechanism, the experimental steps of the BMSC-Exo to high glucose-induced HK2 cell damage protection mechanism are as follows:

[0075] Step 1: grouping and processing of HK2 cells;

[0076] A: Divide HK2 cells into general experimental group and transfection experimental group. The general experimental group includes normal control group, high glucose group, and high osmotic pressure control group. The transfection experimental group includes normal control group, high glucose group, and negative control group. and interference groups;

[0077] B: Put the HK2 cells of the normal control group in the general experimental group into 5.5mM glucose culture solution for cultivation, put the HK2 cells of the high-sugar group in the general experiment group into 25mM glucose culture solution for cultivation, and put the hig...

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Abstract

The invention belongs to the technical field of diabetic nephropathy treatment, and particularly relates to a research method of a protection mechanism of BMSC-Exo on high glucose induced HK2 cell damage. The experimental steps of the protection mechanism of BMSC-Exo on high glucose induced HK2 cell damage include HK2 cell grouping and processing, construction of a miR-886-5p knocked-out HK2 cellline by a Crispr / Cas9 technology, an RNA pull down technology, an FISH experiment, DN establishment and PAS glycogen staining. Bioinformatics prediction is combined with a luciferase reporter vector experiment to prove the interaction between miR-886-5p and a target gene, the RNA pull down experiment and the FISH experiment further prove the binding relationship between miR-886-5 p and the targetgene, and a cure experiment proves that the target gene can reverse the function of miR-886-5p and confirm the mechanism of miR-886-5p in HK2 cells.

Description

technical field [0001] The invention relates to the technical field of diabetic nephropathy treatment, in particular to a research method for the protection mechanism of BMSC-Exo against high glucose-induced HK2 cell damage. Background technique [0002] Pyroptosis is an important factor affecting the inflammatory injury of renal tubular epithelial cells in diabetic nephropathy (DN). The experiment found that bone marrow mesenchymal stem cell-derived exosomes (BMSC-exo) could inhibit high glucose-induced renal tubular epithelial cell (HK2) pyroptosis in vitro. It was found by mass spectrometry that microRNA-886-5p (miR-886-5p) was significantly upregulated in HK2 cells after adding BMSC-exo, suggesting that miR-886-5p may be involved in the occurrence of DN. Therefore, this study will explore the protective mechanism of BMSC-Exo against high glucose-induced HK2 cell damage, and focus on elucidating the inhibitory effect of miR-886-5p on high glucose-induced HK2 cell pyropto...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12Q1/6841G01N1/30G01N33/68A61K49/00
CPCC12Q1/6841C12N15/113G01N1/30G01N33/6869A61K49/0008C12N2310/141C12Q2565/518C12Q2563/107C12Q2543/10
Inventor 吴春铃赵贵芳王芝川葛正鑫李顺康邹平黄珀陈蓉
Owner SOUTHWEST MEDICAL UNIVERISTY
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