Effects Of High Mobility Group Protein 1-interleukin 1 ? Complex On Proliferation,Apoptosis And Autophagy Of Synovial Fibroblasts In Rheumatoid Arthritis

Objective The high mobility group box 1 interleukin-1? complex was used to study the regulation of proliferation,apoptosis and autophagy in rheumatoid arthritis synovial fibroblasts in vitro.Method 1 Isolation,culture and identification of synovial fibroblasts from rheumatoid arthritis.1.1 Non-selective selection of 10 patients with RA,digestive enzyme culture method and tissue block culture method to culture the synovial tissue.1.2 Light microscopy and vimentin immunocytochemical staining were used to identify the cultured samples,and whether they were rheumatoid arthritis synovial fibroblasts and explored their culture methods.2 Effects of high mobility group protein 1-interleukin-1? complex on proliferation of synovial fibroblasts in rheumatoid arthritis.2.1 After PI staining,the changes of S/G2 in each group were analyzed by flow cytometry,and the changes of cell division phase under the action of high mobility group-1 interleukin-1? complex were investigated.2.2 CCK-8 method was used to detect the proliferation of cells in each group,and the changes of cell proliferation under the influence of high mobility group-1 interleukin-1? complex were observed.3 Effects of high mobility group protein 1-interleukin-1? complex on apoptosis of synovial fibroblasts in rheumatoid arthritis.3.1 Annexin V-FITC/PI staining was used to detect apoptosis after flow cytometry,and the change ofapoptotic rate under high mobility group-1 interleukin-1? complex was investigated.3.2 Western blot was used to detect the change of gray scale ratio of Bax and Bcl-2,and to observe the expression of apoptosis-related protein under the influence of high mobility group-1 interleukin-1? complex.4 Effects of high mobility group protein 1-interleukin-1? complex on autophagy of synovial fibroblasts in rheumatoid arthritis.4.1 Western blot was used to detect the difference of LC3 II expression,and the effect of high mobility group-1 interleukin-1? complex on the expression of autophagy protein was investigated.4.2 Immunofluorescence antibody staining and fluorescence microscopy were used to observe the production of autophagy-related protein LC3 II,and to investigate the effect of high mobility group-1 interleukin-1? complex on autophagy protein expression.Result 1 Primary culture was carried out by digestive enzymatic method and tissue block culture method.There was no significant difference between the two methods.Light microscopy and Vimentin immunohistochemical identification and cultured cells were the rheumatoid arthritis required for the experiment.Synovial fibroblasts.2High mobility group protein 1-interleukin-1? complex interfered with rheumatoid arthritis synovial fibroblasts for 48 h,PI staining experimental results and CCK-8 experimental results showed that RASGB1-IL-1? complex group RASFs proliferated significantly The number of cells in S/G2 phase increased significantly,and the difference was statistically significant.3 Annexin V-FITC/PI staining results and the expression of Bax and bcl-2 in Western blot showed no significant changes in the apoptosis of synovial fibroblasts in rheumatoid arthritis group with high mobilitygroup-1 interleukin-1? complex group.,The difference was not statistically significant.4 Western blot and immunofluorescence LC3 B expression showed that there was no significant change in autophagy of rheumatoid arthritis synovial fibroblasts in high mobility group-1 interleukin-1? complex group,and the difference was not statistically significant.Conclusion Under the experimental conditions of this study,high mobility group box-1 interleukin-1? complex promoted rheumatoid arthritis synovial fibroblast proliferation,but had no significant effect on its apoptosis and autophagy.High mobility group protein 1-interleukin 1 beta complex can lead to pathological changes in rheumatoid arthritis synovial fibroblasts and play an important role in the course of RA.