Li Fengfeng, Fan Cunyi, Cheng Tao, Jiang Chaoyin, Zeng Bingfang
Department of Orthopaedics, The Sixth Affiliated People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
Biochem Biophys Res Commun. 2009 May 1;382(2):259-63. doi: 10.1016/j.bbrc.2009.02.165. Epub 2009 Mar 13.
Transforming growth factor-beta1 and fibroblast growth factor-2 play very important roles in fibroblast proliferation and collagen expression. These processes lead to the formation of joint adhesions through the SMAD and MAPK pathways, in which ERK2 is supposed to be crucial. Based on these assumptions, lentivirus (LV)-mediated small interfering RNAs (siRNAs) targeting ERK2 were used to suppress the proliferation and collagen expression of rat joint adhesion tissue fibroblasts (RJATFs). Among four siRNAs examined, siRNA1 caused an 84% reduction in ERK2 expression (p<0.01) and was selected as the most efficient siRNA for use in this study. In subsequent experiments, significant downregulation of types I and III collagen were observed by quantitative RT-PCR and Western blot analyses. MTT assays and flow cytometry revealed marked inhibition of RJATF proliferation, but no apoptosis. In conclusion, LV-mediated ERK2 siRNAs may represent novel therapies or drug targets for preventing joint adhesion formation.
转化生长因子-β1和成纤维细胞生长因子-2在成纤维细胞增殖和胶原蛋白表达中发挥着非常重要的作用。这些过程通过SMAD和MAPK信号通路导致关节粘连的形成，其中ERK2被认为起着关键作用。基于这些假设，使用慢病毒（LV）介导的靶向ERK2的小干扰RNA（siRNA）来抑制大鼠关节粘连组织成纤维细胞（RJATF）的增殖和胶原蛋白表达。在所检测的四种siRNA中，siRNA1使ERK2表达降低了84%（p<0.01），并被选为该研究中最有效的siRNA。在随后的实验中，通过定量RT-PCR和蛋白质印迹分析观察到I型和III型胶原蛋白的显著下调。MTT分析和流式细胞术显示RJATF增殖受到明显抑制，但未出现凋亡。总之，LV介导的ERK2 siRNA可能代表预防关节粘连形成的新疗法或药物靶点。
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