烙灸激活Fas/FasL信号通路对类风湿关节炎大鼠成纤维样滑膜细胞增殖及凋亡的影响OA

Objective It is to investigate the effects of ironing moxibustion(IM)on proliferation and apoptosis of fibro-blast-like synoviocyte(FLS)in rheumatoid arthritis(RA)rats through activation of the tumor necrosis factor receptor su-perfamily member 6(Fas)/Fas-ligand(FasL)signaling pathway.Methods Fifty Sprague-Dawley rats were randomly di-vided into blank group,model group,inhibitor group,IM group and IM+inhibitor group,with 10 rats in each group.The rats in the blank group were injected with the same volume of normal saline like other group into their knee joints,while the rats of the other groups were injected with Freund's complete adjuvant(0.5 mL/kg)to establish RA models.After suc-cessful modeling,the inhibitor group was intraperitoneally injected with Fas/FasL inhibitor(0.1 mg/kg)daily,the IM group was treated with IM at acupoints such as Liangqiu,Xuehai,Yinlingquan,Yanglingquan and Zusanli once daily(2 min per time),the IM+inhibitor group was treated with IM after intraperitoneal injection with Fas/FasL inhibitor0.1 mg/kg,the blank group and model group were not treated with other therapy.After 30 days of continuous treatment,the FLS were isolated and identified by immunofluorescence with Vimentin,their proliferation ability was detected by CCK-8 and EdU staining,and their apoptosis was evaluated by TUNEL staining,the protein and mRNA expressions of Vimentin,CD90,Caspase-3 and Caspase-8 were detected by Western blot and quantitative real-time PCR,respectively.Results Compared with the blank group,the survival rates of FLS and EdU-positive rates of the model group and inhibitor group were significantly increased(all P<0.05),the cell positive rates of TUNEL detection were significantly decreased(both P<0.05),and the relative protein and mRNA expressions of Vimentin,CD90,Caspase-3 and Caspase-8 in FLS were signifi-cantly increased(all P<0.05).Compared with the model group and inhibitor group,the survival rates of FLS and EdU-positive rates of the IM group and IM+inhibitor group were significantly decreased(all P<0.05),the cell positive rates of TUNEL detection were significantly increased(both P<0.05),and the relative protein and mRNA expressions of Vim-entin,CD90 in FLS were significantly decreased(all P<0.05),while the relative protein and mRNA expressions of,Caspase-3 and Caspase-8 were all significantly increased(all P<0.05).Compared with the IM moxibustion group,the a-bove indicators in the IM+inhibitor group were all reversed,and the differences were all statistically significant between the two groups(all P<0.05).Conclusion IM can inhibit the proliferation of FL cells and promote their apoptosis in RA rats via activating the Fas/FasL pathway.