三七总皂苷对过氧化氢诱导的骨髓基质细胞凋亡的保护作用(英文)  

Panax notoginseng saponins protect rabbit bone marrow stromal cells from hydrogen peroxide-induced apoptosis

作　　者：强辉[1,2] 王坤正[1] 张晨[1] 时志斌[1] 樊立宏[1] 

机构地区：[1]西安交通大学医学院第二附属医院骨科,陕西西安710004 [2]陕西省人民医院骨科,陕西西安710068

出　　处：《中西医结合学报》2010年第2期131-137,共7页Journal of Chinese Integrative Medicine

基　　金：国家自然科学基金资助项目(No.30600624)~~

摘　　要：目的:观察三七总皂苷(Panax notoginseng saponins,PNS)对过氧化氢诱导的兔骨髓基质细胞(bone mar-row stromal cell,BMSC)凋亡的影响。方法:从2月龄新西兰兔获得原代BMSC,给予不同浓度PNS处理后,通过检测BMSC增殖能力和碱性磷酸酶活性观察BMSC早期成骨分化能力,筛选出PNS对BMSC作用的最佳浓度。采用过氧化氢(100μmol/L)诱导BMSC凋亡。PNS对过氧化氢诱导前的BMSC进行预处理后,应用2′,7′-二氯荧光黄双乙酸盐法检测细胞活性氧水平,流式细胞术检测细胞凋亡率,免疫印迹法检测BMSC内Bax蛋白水平,荧光分光光度计检测BMSC内半胱氨酸天冬氨酸特异性蛋白酶3(caspase-3)活性。结果:PNS作用的最佳浓度是0.1g/L。与单纯过氧化氢处理组相比,0.1g/LPNS预处理能减少过氧化氢诱导后BMSC内的活性氧含量的升高,降低BMSC凋亡率,减少Bax蛋白表达,并降低caspase-3活性(P<0.01)。结论:PNS可能通过减少氧化应激反应、Bax表达及caspase-3活性发挥其对过氧化氢诱导BMSC凋亡的保护作用。Objective： To investigate the effects of Panax notoginseng saponins （PNSs） on hydrogen peroxide-induced apoptosis in rabbit bone marrow stromal cells （BMSCs）. Methods： BMSCs were isolated from 2-month-old New Zealand rabbits and cultured with different doses of PNSs to determine the most effective dose of PNSs by 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （MTT） assay and alkaline phosphatase （ALP） assay. The most effective dose of PNSs was used in subsequent experiments. Apoptosis of BMSCs was induced by hydrogen peroxide （100 μmol/L）. BMSCs in PNSs group were also pretreated with PNSs before hydrogen peroxide exposure. Reactive oxygen species （ROSs） levels were measured by using 2′,7′-dichlorodihydrofluorescein diacetate. Apoptosis rate of BMSCs was observed by flow cytometry after staining with Annexin V-fluorescein isothiocyanate/propidium iodide. The protein expression of Bax in BMSCs was analyzed by Western blotting. Activity of caspase-3 was measured by spectrofluorometry. Results： The most effective dose of PNSs was 0.1 g/L. PNSs at dose of 0.1 g/L markedly reversed the augmentation of ROS level, decreased the apoptosis rate of, and the Bax expression and activity of caspase-3 in BMSCs treated with hydrogen peroxide （P〈0.01）. Conclusion： PNSs can protect cultured rabbit BMSCs from hydrogen peroxide-induced apoptosis by decreasing oxidative stress, Bax expression and caspase-3 activity.

关 键 词：三七 过氧化氢 骨髓基质细胞 凋亡 兔 

分 类 号：R285[医药卫生—中药学]