纤维样滑膜细胞炎症模型 z-bo 2021-01-05 311

　　Objective To establish a primary culture method of normal knee joint synovial cells and an inflammation model of fibroblast-like synovial cells induced by lipopolysaccharide in rats.

　　Method 80~120g SPF grade juvenile SD rats were selected, their synovial tissues were separated, and primary cultured to the third generation, the FLS protein marker vimentin and proliferation function were detected by histochemical staining method and EdU method. At the same time, synovial tissue was used as a control to detect the characteristic protein expression of FLS from the third to the eighth generation of primary culture, and to screen FLS cells with high purity and physiological functions that can be used in subsequent experiments. After LPS induces FLS inflammation, detect the mRNA and protein expression of IL-1β and TNF-α at different time points when LPS stimulates FLS, and determine the time point when LPS successfully induces FLS inflammation model according to the experimental results. Finally, detect the expression of cytokines, proliferation function and characteristic protein before and after FLS is induced by LPS, and provide experimental data for analyzing FLS inflammation model.

　　 Results Using 0.2% type I collagenase digestion method, FLS primary cells were successfully cultured. After testing the protein marker vimentin, the purity of the third-generation FLS was over 98%. Through FLS characteristic protein detection, FLS with physiological functions and can be used as follow-up experiments was screened out as the 3rd to 7th generation primary cells. By analyzing the cytokines and characteristic proteins of FLS induced by LPS, it is determined that 1μg/mL LPS induces FLS cells for 3h, which can replicate the FLS inflammation model.

　　 Conclusion LPS-induced FLS inflammation model can be used as a cell model for studying inflammatory joint disease in vitro.