钩藤散对NIH-3T3细胞衰老模型增殖与凋亡的影响

目的: 观察钩藤散对NIH-3T3细胞衰老模型增殖与凋亡的影响。方法: NIH-3T3细胞接种在含10%胎牛血清的DMEM培养至20代,然后分为年轻、空白、模型3个组,传至30代时, 用H₂O₂处理,β-半乳糖苷酶染色试剂盒染色,观察衰老细胞形态、衰老指征,确认衰老模型是否成功建立;用钩藤散(17,8.5,4.25,2.15 g ·kg^-1)ig SD大鼠,每天2次,连续给药3 d, 于末次ig 1 h后麻醉、腹主动脉采血,制备含药血清;用钩藤散含药血清处理衰老模型,观察衰老细胞形态、衰老指征、增殖与凋亡等指标。结果: 模型组细胞较对照组形态有显著改变,细胞衰老比例明显增加(P<0.05),因此H₂O₂氧化应激方法可成功建立细胞衰老模型;钩藤散含药血清能明显减少细胞形态的改变、改善衰老指征、促进细胞增殖(P<0.01)、降低细胞凋亡(P<0.01),尤其10倍剂量组最为明显。结论: 钩藤散能有效促进细胞增殖,降低细胞凋亡,从而防止衰老。

Effects of Gouteng San on the Proliferation and Apoptosis of NIH-3T3 Cell Aging Model

Objective: To observe the regulatory effect of Gouteng San on the proliferation and apoptosis of NIH-3T3 cell lines. Method: NIH-3T3 cells were cultivated in DMEM with 10% fetal calf serum and three groups such as youth, blank and model group were used. Cells at generation 30 were treated with hydrogen peroxide (H₂O₂) and then β-galactosidase dyeing kit was applied to observe cell morphology and aging indications to assure whether the aging mode was successfully established.Oral administration of Gouteng San was started for three constitutive days, twice daily. One hour after last administration, rats were anesthetized, and we drew blood from abdominal aorta for the preparation of Gouteng San medicated serum. NIH-3T3 cells were treated with Gouteng San medicated serum to observe cell morphology, aging indications, proliferation and apoptosis. Result: The cell morphology of model was changed and the proportion of cell senescence was increased significantly in model group than in normal control group(P<0.05).Therefore the aging model was successfully established by inducing NIH-3T3 cells aging via oxidative stress with hydrogen peroxide (H₂O₂). Gouteng San medicated serum can significantly attenuate the change of cell morphology, improve aging indications, foster proliferation, (P<0.01) and reduce cell apoptosis(P<0.01), with a distinct effect at 10 times doses. Conclusion: Gouteng San can promote proliferation and reduce apoptosis, thus prevent aging.