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目的 研究羟甲基戊二酸单酰辅酶 A(HMG CoA)还原酶抑制剂辛伐他汀对肺成纤维细胞凋亡的影响。方法 以胎儿肺组织为材料 ,胰蛋白酶消化法培养肺成纤维细胞 ,免疫组化方法鉴定 ,以辛伐他汀 5μM、1 0 μM、2 0 μM及不含辛伐他汀的溶剂作用 72小时 ,另以辛伐他汀 1 0 μM作用2 4、48、72、96小时 ,TUNEL及流式细胞仪定性及定量检测细胞凋亡。结果 (1 )成功培养出肺成纤维细胞 ,光镜下细胞成梭形 ,免疫组化显示细胞表达纤维粘连蛋白和波动蛋白 ,不表达角化蛋白和结合蛋白。 (2 )TUNEL检测结果证实辛伐他汀可诱导细胞凋亡 ,光镜下也显示凋亡细胞的形态改变。流式细胞仪检测结果显示细胞凋亡率随辛伐他汀浓度及作用时间的增加而增加。结论 辛伐他汀能以时间和剂量依赖式诱导肺成纤维细胞凋亡
Objective To investigate the effect of simvastatin, a inhibitor of hydroxymethylglutaryl coenzyme A (HMG CoA) reductase, on the apoptosis of lung fibroblasts. Methods Fetal lung tissue was used as material. Pulmonary fibroblasts were cultured by trypsin digestion and identified by immunohistochemistry. Simvastatin 5 μM, 10 μM, 20 μM and simvastatin without solvent were used for 72 hours. At simvastatin 10 μM for 2, 4, 48, 72 and 96 hours, TUNEL and flow cytometry were used to detect apoptosis. Results (1) The lung fibroblasts were successfully cultured. The cells under the light microscope were spindle-shaped. Immunohistochemistry showed that the cells expressed fibronectin and fluctuating protein, but not keratin and binding protein. (2) TUNEL test results confirmed that simvastatin can induce apoptosis, also showed morphological changes of apoptotic cells under light microscope. Flow cytometry results showed that apoptosis rate increased with the concentration of simvastatin and the effect of time. Conclusion Simvastatin induces lung fibroblast apoptosis in a time-and dose-dependent manner