脱氢表雄酮抗衰老机制的实验研究  

Study on anti-aging mechanisms of dehydroepiandrosterone

作　　者：刘启[1] 赵志英[2] 张凤久[3] 李志辉[1] 

机构地区：[1]包头医学院第二附属医院,内蒙古包头014030 [2]包头医学院,内蒙古包头014000 [3]赤峰学院第一附属医院,内蒙古赤峰024000

出　　处：《现代预防医学》2014年第11期2059-2062,共4页Modern Preventive Medicine

基　　金：内蒙古教育厅高校科研项目(NJ06013)

摘　　要：目的体外培养人胚肺二倍体成纤维细胞(WI-38),从细胞信号转导途径深入研究脱氢表雄酮(Dehydroepiandrosterone DHEA)对细胞生长的影响。方法体外培养人胚肺二倍体成纤维细胞,45代以后细胞应用β-半乳糖甘酶染色方法检测,以阳性细胞比例达到90%以上的细胞为衰老模型;实验分6组,对照组28代,模型组56代,低、中、高剂量组(56代细胞中加入DHEA终浓度分别为5 nmol/ml、50 nmol/ml和100 nmol/ml),抑制剂组(56代细胞中加入终浓度为20 nmol/ml PD98059和100 nmol/ml DHEA);四甲基偶氮唑盐(MTT)法观察细胞活力、流式细胞仪检测各组细胞周期、免疫组化检测细胞外调节蛋白激酶(Extra cellular regulated protein kinases,ERK)蛋白表达变化。结果 (1)各组β-半乳糖苷酶染色阳性细胞率均高于对照组(P<0.01),中剂量组和高剂量组阳性细胞率低于模型组(P<0.05);除高剂量组外其余四组细胞活力均低于对照组(P<0.05),中、高剂量组细胞活力高于模型组(P<0.05);(2)除高剂量组外其余四组G0/G1期细胞比例均高于对照组(P<0.05),中、高剂量组G0/G1期细胞比例低于模型组(P<0.05);除中、高剂量组外其余三组S期细胞比例低于对照组(P<0.05),高剂量组S期细胞比例高于模型组(P<0.05);(3)除高剂量组外,其余四组ERK蛋白表达IOD值均低于对照组(P<0.05),中、高剂量组ERK蛋白IOD值高于模型组(P<0.05)。结论 DHEA能够延缓人胚肺二倍体成纤维细胞衰老,其作用可能与激活ERK信号转导通路有关。Objective To study the effect of the dehydroepiandrosterone on the growth of embryo lung diplont fibroblast in cell signal transduction. Methods Human embryo lung diplont fibroblast were cultured and β -galactosidase staining was used to measure senescent cells after the 45 generation ceils, and the positive cell ratio more than 90% of cell lines was the aging model. There were 6 groups in the research, the control group （28th generations cell）, the model group （56th generations cell）, the low dose group （added DHEA 5 nmol/ml to the 56th generations cell）; the middle dose group （added DHEA 50nmol/ml to the 56th generations cell）, the high dose group （added DHEA 100 nmol/ml to the 56th generations cell）, the inhibitor group （added 20nmol/ml PD98059 and 100 nmol/ml DHEA to the 56th generations ceil）. Cell viability was detected by MTT, and cell cycle was observed by flow cytometry and the immunohistochemistry was used to check the protein expression of ERK in each groups. Results （1） Of each group, β -galactosidase positive cells were higher than that of the control group （/9〈0.01）; Positive ceils of the middle dose group and the high dose group were lower than that of the model group （P〈0.05）; except the high dose group, cell viability of the other four groups were lower than that of the control group （P〈0.05）. Cell viability of the middle dose group and the high dose group were higher than that of the model group （P〈0.05）. （2） Except the high dose group, G0/G1 phase cell percentage of the other four groups of was higher than that of the control group （P〈0.05）, the G0/G1 phase of the middle dose group and the high dose group were lower than that of the model group （P〈0.05）; Except the high dose group and the middle dose group, the percentage in S phase of the other three groups of were lower than that of the control group （P〈0.05）, the percentage of the high dose group in S phase was higher than that of model group （P〈0.05）. （3）

关 键 词：脱氢表雄酮 人胚肺二倍体成纤维细胞 细胞外调节蛋白激酶 衰老 

分 类 号：R113[医药卫生—公共卫生与预防医学]