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目的探讨RNA干扰(RNA interference,RNAi)介导的蛋白酶体亚基RC2(Rat component 2)下调延缓失神经肌萎缩的疗效。方法将48只SD大鼠,建立右下肢趾长伸肌失神经支配模型,随机分为实验组和对照组,每组24只。在电穿孔技术介导下,实验组和对照组分别转染含RC2基因和非同源对照序列的siRNA重组质粒溶液(0.8μg/μl)各50μl。转染后3 d,采用Western blot测定两组趾长伸肌中RC2蛋白含量;转染后14、21、28 d,测定肌湿重维持率、肌肉蛋白含量和肌纤维横截面积,并观察肌纤维超微结构的变化。结果转染后3 d,实验组RC2的蛋白水平明显下调,抑制率为25%,与对照组相比差异有统计学意义(P<0.05);转染后14 d和21 d实验组肌湿重维持率、肌肉蛋白含量和肌纤维横截面积均明显优于对照组(P<0.05);28 d时,两组间各项指标差异无统计学意义(P>0.05)。结论RNA干扰介导的蛋白酶体亚基RC2下调延缓失神经肌萎缩的疗效可以维持3周。
Objective To investigate the effect of RNA interference (RNAi) -mediated inhibition of rat component 2 (RC2) on denervation and amyotrophy. Methods Forty eight SD rats were divided into experimental and control groups, 24 rats in each group. Under electroporation, 50μl of siRNA recombinant plasmid solution (0.8μg / μl) containing RC2 gene and non-homologous control sequences were transfected into the experimental group and the control group respectively. At 3 d after transfection, the RC2 protein content in the extensor digitorum longus was measured by Western blot. At 14, 21 and 28 days after transfection, the maintenance of muscle wet weight, muscle protein content and muscle fiber cross-sectional area were measured, Changes in ultrastructure. Results At 3 days after transfection, the protein level of RC2 was significantly down-regulated in the experimental group (25%), which was significantly different from that of the control group (P <0.05). On the 14th and 21st days after transfection, the muscle wetness The weight retention, muscle protein content and muscle fiber cross-sectional area were significantly better than those of the control group (P <0.05). There was no significant difference between the two groups on the 28th day (P> 0.05). Conclusion The RNA interference-mediated proteasome subunit RC2 down-regains the function of denervation and amyotrophy for 3 weeks.