RNA干扰沉默STAT3对胶原诱导大鼠类风湿性关节炎的治疗作用

doi:10.13481/j.1671-587x.20150115

摘要/Abstract

摘要：

目的: 研究小干扰RNA(siRNA)干扰沉默信号转导和转录激活因子3(STAT3)对大鼠类风湿关节炎(RA)的治疗作用,评价其有效性,为RA的基因治疗提供实验依据。方法: 采用鸡Ⅱ型胶原诱导方法建立大鼠 RA(CIA)模型,实验分为正常组(非关节炎大鼠)、模型组(不给予治疗的关节炎大鼠)、阴性对照组(给予乱码RNA的关节炎大鼠)和治疗组(给予STAT3特异干扰siRNA的关节炎大鼠),实验中siRNA均由慢病毒颗粒包载。实验期间观察大鼠一般情况,每7 d检测大鼠关节病变程度并记录,实验第35天处死大鼠后观察其关节病理切片并进行评分;Western blotting法检测大鼠滑膜组织中STAT3蛋白表达水平;RT-PCR法检测大鼠滑膜组织中STAT3 mRNA表达水平。结果: 与模型组和阴性对照组比较,治疗组大鼠临床及形态学表现均明显减轻。与正常组比较,模型组和阴性对照组大鼠滑膜组织中STAT3 mRNA和蛋白表达水平明显增加(P<0.05或P<0.01),治疗组差异无统计学意义(P>0.05);模型组与阴性对照组比较差异无统计学意义(P>0.05)。与模型组比较,治疗组大鼠滑膜组织中STAT3 mRNA和蛋白表达水平明显下降(P<0.05)。结论: 局部注射siRNA阻断STAT3的表达可明显改善RA大鼠关节的组织病理学表现,其机制可能与STAT3 mRNA和蛋白表达水平降低有关。

关键词: 类风湿关节炎, RNA干扰, 信号转导和转录激活因子3, 慢病毒载体

Abstract:

Objective To study the therapeutic effect of small interfering RNA (siRNA) interference to silence STAT3 on rheumatoid arthritis (RA) in the rats, to evaluate the effectiveness, and to provide experimental basis for the gene therapy of RA. Methods The RA models of rats(CIA) were constructed by chickenⅡcollagen induced method.There were four groups in this experiment: normal group (the rats without arthritis), model group (the rats with arthritis but without treatment), negative control group (the rats with RA with messy coded RNA) and treatment group (the rats with RA with STAT3 specifically coded interference siRNA ).Both siRNA were entrapped into lentiviral particles and were injected into the rats' joints.The general situations of the rats were observed and the pathological degrees of the joints were recorded every 7 days and the pathological section of the rat joints were observed;the rats were sacrificed on the 35th day and the pathological sections of the joints were scored.RT-PCR method was used to detect the STAT3 mRNA expression levels after siRNA interference and the expression levels of STAT3 protein in synovial tissue of the rats were detected by Western blotting method. Results The clinical and morphological appearance of the rats in treatment group was significantly lightened compared with model group and negative control group.Compared with normal group, the expression levels of STAT3 mRNA and protein in synovial tissue in model group and negative control group were significantly increased(P<0.01), and the expression levels of STAT3 mRNA and protein in treatment group had no significant difference(P>0.05);there was no significant difference between model group and negative control group (P>0.05).Compared with model group, the expression levels of STAT3 mRNA and protein in synovial tissue of the rats in treatment group were significantly decreased (P<0.05). Conclusion Local injection of siRNA can block the expression of STAT3, and thus can make significant improvements in pathological manifestations in the joints of RA rats.The mechanism may be related to the reduction of the expression levels of STAT3 mRNA and protein.

Key words: rheumatoid arthritis, RNA interference, signal transducer and activator of transcription 3, lentiviral vector

中图分类号:

R593.22

刘畅, 高薇, 冯欣. RNA干扰沉默STAT3对胶原诱导大鼠类风湿性关节炎的治疗作用[J]. 吉林大学学报(医学版), 2015, 41(01): 77-82.

LIU Chang, GAO Wei, FENG Xin. Therapeutic effect of RNA interfered STAT3 silence on collagen induced rheumatoid arthritis in rats[J]. Journal of Jilin University Medicine Edition, 2015, 41(01): 77-82.

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