动物造模,成纤维样滑膜细胞炎症模型 z-bio 2021-09-23 700

　　OBJECTIVE: To establish a primary culture of normal knee joint synovial cells in Sprague Dawley (SD) rats and a lipopolysaccharide (LPS)-induced fibroblast-like synovial cell inflammation model.

　　Method: 80-120 g SPF grade immature SD rats were selected, synovial tissues were separated, and primary cultured to the third generation, and the FLS protein markers were detected by histochemical staining and EdU method for vimentin and proliferation function. At the same time, using synovial tissue as a control to detect the characteristic protein expression of FLS from the 3rd to 8th generation of primary culture, it has high purity and physiological function and can be used in subsequent experiments. Be screened. After LPS induces FLS inflammation, detect the mRNA and protein expression of IL-1β and TNF-α at different times when LPS stimulates FLS, and judge when LPS successfully induces FLS inflammation model based on the experimental results. Finally, we tested the expression of cytokines, proliferation function and characteristic proteins before and after LPS induced FLS, and provided experimental data for the analysis of FLS inflammation models.

　　Result: We successfully cultivated FLS primary cells using 0.2? collagenase digestion method. After the detection of protein-labeled vimentin, the purity of the third-generation FLS reached 98?. Due to the protein detection characteristics of FLS, FLS, which has physiological functions and can be used as a follow-up experiment, is selected as the 3-7 generation primary battery. Analysis of LPS-induced FLS cytokines and characteristic proteins confirmed that 1μg/mL LPS induced FLS cells for 3 hours, which can replicate the FLS inflammation model.

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