摘要
目的:探讨RNA干扰沉默TLR4对类风湿关节炎(RA)滑膜成纤维细胞增殖和侵袭的影响。方法:将体外培养的人RA滑膜成纤维细胞MH7A分为对照组(未转染)、NC-siRNA组(转染阴性对照siRNA)和TLR4-siRNA组(转染TLR4-siRNA)。采用Western blot法检测3组细胞中TLR4、PCNA、MMP-9和VEGF蛋白的表达,实时荧光定量PCR法检测TLR4 mRNA的表达,MTT法检测细胞增殖活性,克隆形成实验检测细胞克隆形成能力,Transwell小室法检测细胞侵袭能力。结果:与对照组和NC-siRNA组比较,TLR4-siRNA组细胞中TLR4、PCNA、MMP-9、VEGF蛋白和TLR4 mRNA的表达均降低(P <0. 05),细胞增殖活性、克隆形成能力和侵袭能力减弱(P <0. 05)。结论:沉默TLR4可通过下调PCNA、MMP-9和VEGF表达抑制RA滑膜成纤维细胞的增殖和侵袭。
Aim: To evaluate the effects of TLR4 silencing induced by RNA interference on proliferation and invasion of rheumatoid arthritis( RA) synovial fibroblasts. Methods: Human RA synovial fibroblast cells MH7 A were divided into control group( untransfected),NC-siRNA group( transfected with NC-siRNA) and TLR4-siRNA group( transfected with TLR4-siRNA). The expressions of TLR4,PCNA,MMP-9 and VEGF proteins were detected by Western blot,the expression of TLR4 mRNA was measured by qRT-PCR,the proliferation activity was tested by MTT,the clonal formation ability was checked by clone formation assay,and the invasion ability was examed by Transwell chamber assay. Results: Compared with the control group and NC-siRNA group,the expressions of TLR4,PCNA,MMP-9,VEGF proteins and TLR4 mRNA in TLR4-siRNA group decreased( P < 0. 05),and the proliferation activity,clonal formation and invasion abilities in TLR4-siRNA group were significantly weakened( P < 0. 05). Conclusion: TLR4 silencing can inhibit the proliferation and invasion of RA synovial fibroblasts by down-regulating the expressions of PCNA,MMP-9 and VEGF.
作者
鲍晓
何成松
BAO Xiao;HE Chengsong(Department of Rheumatology and Immunology,the Affiliated Hospital of Southwest Medical University,Luzhou,Sichuan 646000;Department of Rheumatology and Immunology,Deyang People's Hospital,Deyang,Sichuan 618000)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2019年第2期187-190,共4页
Journal of Zhengzhou University(Medical Sciences)
基金
四川省科学技术厅泸州市人民政府泸州医学院联合专项资金计划项目(川科发计201410074)
