Protective effects and mechanism of Panax Notoginseng saponins on oxidative stress-induced damage and apoptosis of rabbit bone marrow stromal cells
Protective effects and mechanism of Panax Notoginseng saponins on oxidative stress-induced damage and apoptosis of rabbit bone marrow stromal cells
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DOI：
10.1007/s11655-010-0566-1
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发表时间：
2010-12-01
影响因子：
2.9
通讯作者：
Wang Kun-zheng
中科院分区：
文献类型：
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作者：
Qiang Hui;Zhang Chen;Wang Kun-zheng
探讨三七皂苷（PNS）对氧化应激诱导的骨髓基质细胞（BMSCs）损伤和凋亡的影响及其可能机制。采用密度梯度离心结合贴壁法从2月龄新西兰兔身上分离培养骨髓间充质干细胞。第三代细胞用于后续实验。0.1 mmol/L的H2O2诱导骨髓间充质干细胞氧化应激。在H2O2处理前，用25-200 μ g/mL PNS预处理骨髓间充质干细胞4 h。MTT法观察骨髓间充质干细胞的增殖情况。碱性磷酸酶（ALP）活性作为早期成骨细胞分化的指标，采用碱性磷酸酶测定试剂盒进行测定。annexv - fitc /碘化丙啶染色，采用流式细胞术观察骨髓间充质干细胞凋亡情况。采用活性氧（ROS）法检测氧化应激水平。Western blotting检测骨髓间充质干细胞中Bax、Bcl-2和Caspase-3蛋白的表达。PNS对H2O2诱导的骨髓间充质干细胞增殖和成骨分化具有不同浓度依赖性。确定PNS浓度为100 μ g/mL为最佳有效浓度。PNS可显著降低h2o2诱导的细胞凋亡率，由41.91%降至14.67% （P < 0.01）。PNS显著降低H2O2诱导的ROS水平（P < 0.01）。此外，PNS预处理显著逆转h2o2诱导的Bcl-2表达抑制和Bax和Caspase-3表达升高（P < 0.01）。PNS通过清除ROS和调节Bcl-2/Bax通路，对氧化应激诱导的兔骨髓间充质干细胞损伤和凋亡具有保护作用。
To investigate the effects and possible mechanism of Panax Notoginseng saponins (PNS) on oxidative stress-induced damage and apoptosis in bone marrow stromal cells (BMSCs).BMSCs were isolated and cultured from 2-month-old New Zealand rabbits by the density gradient centrifugation combined with adherent method. The third passage cells were used for subsequent experiments. Oxidative stress was induced in cultured BMSCs by H2O2 (0.1 mmol/L). BMSCs were pretreated with 25-200 mu g/mL PNS for 4 h before H2O2 treatment. Proliferation of BMSCs was observed using MTT assay. Alkaline phosphatase (ALP) activity, as an index of early osteoblastic differentiation, was determined with an ALP assay kit. Flow cytometry was used to observe the apoptosis of BMSCs by staining with annexinV-FITC/propidium iodide. Oxidative stress level was examined by reactive oxygen species (ROS) assay. The protein expressions of Bax, Bcl-2 and Caspase-3 in BMSCs were analyzed by Western blotting.PNS had different concentration-dependent effects on proliferation and osteoblast differentiation of BMSCs induced by H2O2. A PNS concentration of 100 mu g/mL was determined as the optimal effective concentration. PNS markedly attenuated H2O2-induced apoptosis rate from 41.91% to 14.67% (P < 0.01). PNS significantly decreased ROS level induced by H2O2 (P < 0.01). Furthermore, pretreatment with PNS significantly reversed H2O2-induced inhibition of Bcl-2 expression and augmentation of Bax and Caspase-3 expression (P < 0.01).PNS had a protective effect on oxidative stress-induced damage and apoptosis in cultured rabbit BMSCs through scavenging ROS and regulating the Bcl-2/Bax pathway.