目的 研究蜂毒素(melittin)对人胚肺成纤维细胞(MRC-5)生长的抑制作用及其机制.方法 采用MTT法检测蜂毒素对MRC-5细胞的增殖抑制作用;流式细胞仪检测细胞周期;RT-PCR 法检测细胞中Cyclin D1、CDK4及E2F-1 mRNA的表达变化;Western blot法检测Cyclin D1、CDK4、p21及E2F-1蛋白表达.结果 与对照组比较,蜂毒素处理组细胞增殖明显受到抑制,并呈剂量和时间依赖性(P<0.05);流式细胞仪检测结果发现,随着浓度的增大,G0/G1期细胞百分比逐渐上升,S期细胞百分比逐渐下降;同时Western blot结果提示蜂毒素(1,2,4 mg·L-1)可以明显降低Cyclin D1、CDK4及E2F-1表达,而上调p21表达.结论 蜂毒素能抑制MRC-5细胞增殖,其机制可能与干扰该细胞G0/G1期相关基因的转录相关且抑制Cyclin D1/E2F信号转导.%Aim To investigate the inhibitory effect of Melittin on the growth of human lung fibroblasts, and its mechanisms. Methods The inhibition rate of growth of cells treated with Melittin was evaluated by MTT assay. The cell cycle distribution was detected by flow cytometric analysis. The expressions of Cyclin D1, CDK4 and E2F-1 mRNA in MRC-5 cells were detected by RT-PCR. The protein expressions of Cyclin Dl , CDK4 and E2F-1 were observed by Western blot. Results Compared with the control group, Melittin had obvious inhibitory effect on the growth of human lung fibroblasts in a dose-and-time dependent manner. After the treatment with Melittin at different concentrations for 24 h, the percentage of the cells in G0/G1 phase increased while in S phase decreased ( P < 0. 05 ). The results showed that Melittin( 1,2,4 mg · L -1)could reduce the expressions of Cyclin D1 , CDK4 and E2F-1( P <0. 05 )but up-regulate the expression of p21. Conclusions Proliferation activity of MRC-5 cells is inhibited by Melittin, which may be related to the inhibitory effect of Melittin on associated protein transcription and Cyclin D1/E2F signaling transduc-tion.
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