RNAi对基质金属蛋白酶13在兔骨性关节炎中表达的抑制作用

目的 观察RNA干扰技术(RNAi)是否能有效抑制兔OA模型膝关节软骨中基质金属蛋白酶13(MMP13)的表达.方法 体外化学合成MMP13序列特异性双链RNA与脂质体混合后转染OA模型中软骨细胞,实验分四组,特异性RNAi组:特异性RNAi+Lipofectamine2000,非特异RNAi组:非特异RNAi+Lipofectamine2000,脂质体组:Lipofectamine2000,对照组:无血清DMEM.用逆转录-聚合酶链反应(RT-PCR)和免疫组织化学法检测软骨细胞中的MMP13的表达.噻唑蓝(MTT)比色法检测各组的软骨细胞生长情况.结果 RT-PCR显示RNAi组较对照组、非特异性组和脂质体组的MMP13基因的PCR扩增条带明显减弱,免疫组织化学法显示RNAi组中MMP13阳性细胞数比对照组明显减少.与对照组比较,非特异性组、RNAi组、脂质体组软骨细胞的存活率显著降低(P<0.05),但在3组间两两差异无统计学意义.结论 使用RNA干扰技术可有效抑制兔OA模型中膝关节软骨细胞MMP13的表达,RNA干扰技术为基因治疗OA提供了新策略.

Inhibitory effects of RNA interference on the expression of MMP13 in the ehondrocyte of rabbit with osteoarthritis

Objective To explore whether RNA interference (RNAi) could suppress the expres-sion of MMPI3 in the chondrocytes of rats with osteoarthritis (OA). Methods Chondrocytes in OA mod-els were transfected using in vitro chemically-synthesized MMP13 sequence-specific double-stranded RNA formulated with liposome. The experiments were conducted in 4 groups:group A (control group), serum-free DMEM; group B (RNAi group) ,RNAi and Lipfectamine2000; group C (non-distinctive group) ,non-specificity dsRNA and Lipfectamine2000 ; group D (liposome group), Lipofectamine2000. The expression of MMP13 in ehondrocytes was detected by RT-PCR and immunohistochemistry. The proliferation of chron-drocytes was assessed by MTT assay. Results The expression of MMP13 mRNA was decreased markedly in group B as compared with group A, group C and group D. Immunohistochemistry showed the significant decreases of MMP13 in group B as compared with group. MTT results revealed that the growth of chrondro-cytes was suppressed in all 4 groups, but there was no significant difference among the groups A, B and C (P > 0.05). Conclusion RNAi was able to effectively suppress the expression of MMPI3 in chrondro-cytes of rabbit OA models.