动物造模,膝关节成纤维样滑膜细胞炎症模型 z-bio 2021-10-07 804

　　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 isolated, and primary cultured to the third generation, the FLS protein markers were labeled with vimentin by histochemical staining and EdU method, and the proliferation function was detected. 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, the mRNA and protein expressions of IL-1β and TNF-α were detected at different times when LPS stimulated FLS, and judged when LPS successfully induced the 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, was selected as the 3-7 generation primary battery. The analysis of FLS cytokines and characteristic proteins induced by LPS showed that 1μg/mL LPS can induce FLS cells for 3 hours and 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.