论文部分内容阅读
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 observe the effect of panax notoginseng saponins (PNS) on apoptosis induced by hydrogen peroxide in bone marrow stromal cell (BMSC). Methods: Primary BMSCs were obtained from 2-month old New Zealand rabbits. After treated with different concentrations of PNS, the early osteogenic and differentiation ability of BMSC was observed by detecting the proliferation and alkaline phosphatase activity of BMSCs. The optimal concentration of PNS for BMSCs was screened out. BMSC apoptosis was induced by hydrogen peroxide (100μmol / L). After PNS pretreated BMSC before hydrogen peroxide induction, reactive oxygen species (ROS) levels were detected by 2 ’, 7’-dichlorofluorescein diacetate assay, flow cytometry was used to detect the apoptotic rate, and Western blotting was used to detect BMSC Bax protein level. The activity of caspase-3 in BMSC was detected by fluorescence spectrophotometer. Results: The best concentration of PNS was 0.1 g / L. Compared with pure hydrogen peroxide treatment, 0.1g / LPNS pretreatment could reduce the increase of active oxygen in BMSC induced by hydrogen peroxide, decrease the apoptosis rate of BMSC, decrease the expression of Bax and decrease the activity of caspase-3 (P <0.01). CONCLUSION: PNS may exert its protective effect on H2O2-induced BMSC apoptosis through decreasing oxidative stress reaction, Bax expression and caspase-3 activity.