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

作　　者：杨效宁[1] 孙一公[1] 李秀霞[1] 戴醒明[1] 龚亚辉[1] 孔清泉[2] 

机构地区：[1]徐州市第一人民医院骨科,221002 [2]四川大学华西医院骨科

出　　处：《黑龙江医药》2013年第3期385-389,共5页Heilongjiang Medicine journal

基　　金：徐州市2010年科技计划项目(课题编号:XF10C035)

摘　　要：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.

关 键 词：软骨细胞肥大分化 核心结合因子 RNA干扰 腺病毒 

分 类 号：R684[医药卫生—骨科学]