腺病毒介导的RNAi抑制Cbfa1表达阻断软骨细胞肥大分化的实验研究

Advenvirus-mediated Transfer of siRNA against Cbfa1 Inhibits the HyperTrophic Differentiation of Chondrocytes

目的:探讨利用RNA干扰(RNA interference,RNAi)技术来抑制核心结合因子(core binding factor α1,Cbfa1)表达,从而阻断软骨细胞肥大分化目的。方法:构建沉默SD大鼠核心结合因子基因的腺病毒(Ad-Cbfa1-siRNA)。利用维甲酸及IL-1α促进软骨细胞肥大分化,研究Ad-Cbfa1-siRNA对Cbfa1表达的抑制作用。利用Ⅱ型胶原、Ⅹ型胶原免疫组化的方法以及RT-PCR比较各组之间Cbfa1的表达情况来分析软骨细胞的肥大分化情况。结果:结果显示,软骨细胞经维甲酸和IL-1α诱导后,Cbfa1的表达量明显增加,经统计学分析,P<0.05,具有统计学意义。阴性对照病毒组Cbfa1的表达量明显高于Ad-Cbfa1-siRNA组,经统计学分析,P<0.05,具有统计学意义。结论:利用RNAi技术能够明显的抑制Cbfa1的表达,从而阻断软骨细胞的肥大分化。

Objective：We used RNAi technology to suppress Cbfal expression, to achieve the purpose of blocking the hypertrophic diferentiation of chondrocytes Method：We constructed adenovirus containing siRNA against Cbfal（Ad-Cbfal-siRNA）. We used retinoic acid and IL-1 α to induce hypertrophic differetiation of chondrocytes, and then We used Ad-Cbfal-siRNA to inhibits the hypertrophic differentiation of chondrocytes .We used immunohistochemistry to detect the type Ⅱ and type X collagen and RT-PCR to analyze the ex-pression of Cbfal. Results：The result of RT-PCR showed thatthe expression of Cbfal was significantly increased after using the retinoic acid and IL-1 α .The expression of Cbfal in Control virus group was significantly higher than Ad-Cbfal-siRNA group. Conclusion：In vitro,it is likely that the inhibition of Cbfal by RNAi could be a powerful way to prevent the hypertrophic differentiation of chondrocytes.