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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H2 O2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。
Objective: To observe the effects of Gouteng San on the proliferation and apoptosis of NIH-3T3 cells. Methods: NIH-3T3 cells were inoculated in DMEM containing 10% fetal bovine serum for 20 generations and then divided into 3 groups: young, blank and model. After passage 30, the cells were treated with H2O2, β-galactosidase Staining kit, observe the morphology of senescent cells, indications of aging, to confirm the establishment of the aging model successfully; with Uncaria (17,8.5,4.25,2.15 g.kg-1) ig SD rats twice a day, continuous Drug 3 d, in the last ig 1 h after anesthesia, abdominal aorta blood collection, preparation of drug-containing serum; with gouteng San sebaceous serum treatment aging model, observation of senescent cell morphology, aging indications, proliferation and apoptosis indicators. Results: The morphology of cells in the model group changed significantly compared with that of the control group, and the percentage of cell senescence increased significantly (P <0.05). Therefore, H2O2 oxidative stress could establish a cell aging model successfully. , Improve the indications of aging, promote cell proliferation (P <0.01), reduce apoptosis (P <0.01), especially in the 10-fold dose group. Conclusion: Gouteng San can effectively promote cell proliferation, reduce apoptosis and prevent aging.