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目的 :探讨人参皂苷Rg1对抗三丁基过氧化氢 (t BHP)诱导的WI 38细胞衰老作用及其可能细胞周期调控机制。方法 :将WI 38细胞随机分为 4组 ,用不同剂量Rg1预处理。从 30代开始 ,隔代用t BHP作用 ,每次 1h ,共 4次 ,诱导细胞衰老。从光镜、透射电镜观察细胞形态及超微结构 ;流式细胞术分析G1期细胞比例 ;以及SA β 半乳糖苷酶的细胞化学染色 ,确定Rg1的抗衰老作用。并采用免疫印迹技术对CDK4、cyclinD1和p16等表达情况进行检测。结果 :Rg1预处理组与单纯t BHP处理组相比 ,细胞形态体积小、胞体不如后者扁平 ,次级溶酶体减少 ,G1期细胞比例下降 ,SA β 半乳糖苷酶染色阳性细胞百分比下降 ,说明Rg1在t BHP诱导细胞衰老模型中有抗衰老作用。进一步发现用Rg1预处理后 ,p16、cyclinD1表达水平降低、CDK4表达水平增加。结论 :提示Rg1可能通过改变细胞周期调控因子的表达而发挥其抗t BHP诱导的WI 38细胞衰老作用。
Objective: To investigate the effect of ginsenoside Rg1 against the senescence of WI 38 cells induced by tributyl hydroperoxide (t BHP) and its possible cell cycle regulation mechanism. Methods: WI 38 cells were randomly divided into 4 groups and pretreated with different doses of Rg1. From the 30th generation onwards, every other generation was treated with t BHP for 1 h each time for a total of 4 times to induce cell senescence. The morphology and ultrastructure of the cells were observed by light and transmission electron microscopy; the proportion of cells in the G1 phase was analyzed by flow cytometry; and the cytochemical staining of SA beta galactosidase was used to determine the anti-aging effects of Rg1. Immunoblotting was used to detect the expression of CDK4, cyclinD1 and p16. RESULTS: In the Rg1 pretreatment group, compared with the t BHP treatment group, the cell morphology was small, the cell bodies were less flat than the latter, the secondary lysosomes decreased, the proportion of G1 phase cells decreased, and the percentage of SA beta galactosidase staining positive cells decreased. , indicating that Rg1 has anti-aging effects in t BHP-induced cell senescence model. It was further found that after pretreatment with Rg1, the expression level of p16, cyclinD1 was decreased, and the expression level of CDK4 was increased. Conclusions: It is suggested that Rg1 may exert its anti-BHP-induced senescence of WI 38 cells by changing the expression of cell cycle regulators.