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目的:研究沉默Cx43基因对左归丸含药血清调控MC3T3-E1细胞成骨相关基因Runx2、Osterix表达的影响。方法:采用慢病毒介导的RNA干扰技术沉默Cx43基因,培养细胞分为正常对照(A)组、基因沉默(B)组、含药血清(C)组、中药加基因沉默(D)组。RT-PCR法检测培养第7、14、21天细胞Runx2、Osterix m RNA表达情况,进行组间对比。结果:左归丸含药血清可以促进MC3T3-E1细胞分化,显著增加分化中后期Runx2,尤其是Osterix m RNA的表达,A、C组比较,P<0.05或P<0.01。沉默Cx43基因导致细胞分化中后期Runx2、Osterix m RNA表达显著下降,A、B组比较,P<0.05;同时导致左归丸上述作用显著下降,C、D组比较,P<0.05。结论:沉默Cx43基因导致左归丸含药血清促MC3T3-E1细胞Runx2、Osterix m RNA表达的作用显著下降。
Objective: To investigate the effects of silencing Cx43 gene on osteoblast-related genes Runx2 and Osterix in MC3T3-E1 cells mediated by Zuoguiwan. Methods: Cx43 gene was silenced by lentivirus-mediated RNA interference technique. The cultured cells were divided into normal control group (A), gene silencing (B) group, drug-containing serum group (C) and Chinese medicine plus gene silencing (D) group. The expression of Runx2 and Osterix m RNA in cultured cells on days 7, 14, and 21 were detected by RT-PCR. Results: Zuogui pill containing serum could promote the differentiation of MC3T3-E1 cells and significantly increase the expression of Runx2, especially Osterix m RNA in the middle and late stages of differentiation. P <0.05 or P <0.01 in A and C groups. Silencing Cx43 gene resulted in a significant decrease of Runx2 and Osterix m RNA expression in late stage of cell differentiation, P <0.05 in group A and B, and significant decrease of Zuogui pill in the above two groups, P <0.05. CONCLUSIONS: Silencing Cx43 gene results in significantly decreased expression of Runx2 and Osterix m RNA in Zuogui pill-containing MC3T3-E1 cells.