Metabolism-related fatty liver disease in-vitro cell model and construction method thereof

Abstract

The invention belongs to the technical field of cell model construction, and discloses a metabolism-related fatty liver disease in-vitro cell model and a construction method thereof.HepG2 and LO2 cells are treated with OA or PA or a mixture thereof respectively, and the cell survival rate is detected with CCK8; detecting the lipid accumulation degree of the cells by oil red O staining and a triglyceride enzyme method; according to the present invention, qRT-PCR is adopted to detect the expression levels of apoptosis-related proteins Bax, Bcl2 and Cleved caspase-3, fibrosis-related proteins alpha-SMA and Col.I, autophagy-related proteins LC3-II, P62 / SQSTM1 and Beclin-1, and inflammatory factors NLRP3 and TNF-alpha, the saturated fatty acid PA is used for treating HepG2 cells, so that a certain degree of lipid accumulation can be caused, remarkable cell damage and fibrosis are induced, and the MAFLD in-vitro cell model is a proper MAFLD in-vitro cell model.

Description

technical field [0001] The invention belongs to the technical field of cell model construction, and in particular relates to an in vitro cell model of metabolism-related fatty liver disease and a construction method thereof. Background technique [0002] At present, metabolic dysfunction-associated fatty liver disease (MAFLD), formerly known as nonalcoholic fatty liver disease (NAFLD), as scholars gradually deepen their research and understanding of NAFLD, its development may be more or less The latest international expert group consensus mentioned that MAFLD should be an independent inclusion disease rather than an exclusion disease. It is recommended to replace the previous classification of simple fatty liver with liver inflammation activity and fibrosis. and steatohepatitis dichotomous rules to describe the disease process. In the current clinical research, due to the large heterogeneity of the main driving factors and disease regulating factors in the MAFLD population,...

AI Technical Summary

Problems solved by technology

[0004] Through the above analysis, the problems and defects of the prior art are: in the prior art, how PA and OA promote the development of steatosis and cytotoxicity in liver cell lines and the influence on cell damage, fibrosis and inflammation have not been fully understood. describe

[0005] The difficulty in solving the above problems and defects is: the prior art mainly focuses on the establishment of a simple steatosis model to study the occurrence and development of MAFLD, but there is a lack of advanced MAFLD cell models such as steatosis, liver cell damage and fibrosis.

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