摘要
背景:研究证实,人参皂苷Rg1对血管内皮细胞具有保护作用,但其作用机制尚不明确。目的:探讨人参皂苷Rg1对过氧化氢诱导的大鼠骨髓来源内皮祖细胞损伤的保护作用及机制。方法:采用直接贴壁法分离培养SD大鼠骨髓来源内皮祖细胞。将对数生长期的内皮祖细胞分为3组:空白对照组,不进行任何干预,常规培养24 h;细胞损伤组,加入含100μmol/L过氧化氢的EMG-2培养基培养24h;人参皂苷Rg1组,先以64μmol/L人参皂苷Rg1预先干预2h,然后加入含100μmol/L过氧化氢的EMG-2培养基培养22 h。采用CCK-8、Tunel、划痕实验检测细胞增殖、细胞凋亡及迁移能力,Western Blot检测细胞内Akt、p-Akt、Bax、Bcl-2蛋白的表达,ELISA法检测细胞内超氧化物歧化酶、丙二醛、一氧化氮水平。结果与结论:(1)与细胞损伤组比较,人参皂苷Rg1组细胞存活率明显增高(P <0.05),细胞凋亡数量明显降低(P <0.05),细胞迁移能力明显提高(P <0.05);(2)与细胞损伤组比较,人参皂苷Rg1组超氧化物歧化酶、一氧化氮水平明显升高,丙二醛水平显著降低(P均<0.05);(3)3组间Akt蛋白表达无差异,人参皂苷Rg1组p-Akt蛋白表达高于细胞损伤组(P <0.05);(4)人参皂苷Rg1组Bcl-2蛋白表达高于细胞损伤组(P <0.05),Bax蛋白表达低于细胞损伤组(P <0.05);(5)结果表明,人参皂苷Rg1能对抗过氧化氢诱导的大鼠骨髓来源内皮祖细胞损伤,提高细胞活力,抑制细胞凋亡,降低氧化应激损伤,Akt信号通路在其中可能发挥了一定作用。
BACKGROUND:Studies have confirmed that ginsenoside Rg1 has protective effects on vascular endothelial cells,but its underlying mechanism is still unclear.OBJECTIVE:To investigate the mechanism by which ginsenoside Rg1 protects against hydrogen peroxide induced injury of rat bone marrow-derived endothelial progenitor cells.METHODS:Endothelial progenitor cells derived from bone marrow of Sprague-Dawley rats were isolated and cultured by direct adherent method.The endothelial progenitor cells in logarithmic growth period were divided into three groups:control group,no intervention but conventional culture for 24 hours;cell injury group,cells were cultured in EMG-2 medium containing 100μmol/L hydrogen peroxide for 24 hours;combined intervention group,cells were pretreated with ginsenoside Rg1(64μmol/L)for 2 hours,then cultured in EMG-2 medium containing 100μmol/L hydrogen peroxide for 22 hours.Cell proliferation,apoptosis and migration were detected using cell counting kit-8,TUNEL,and cell scratch test,respectively.The expression of Akt,p-Akt,Bax,and Bcl-2 proteins was measured using western blot assay.The levels of intracellular superoxide dismutase,malondialdehyde and nitric oxide were determined using ELISA method.RESULTS AND CONCLUSION:(1)Compared with the cell injury group,the cell survival rate and cell migration ability were significantly increased,while the number of apoptotic cells reduced significantly in the combined intervention group(all P<0.05).(2)Compared with the cell injury group,the levels of intracellular superoxide dismutase and nitric oxide were significantly increased,while the level of malondialdehyde was significantly reduced in the combined intervention group(all P<0.05).(3)No significant difference in the expression of Akt protein was detected among the three groups,but the expression of p-Akt protein was significantly higher in the combined intervention group than the cell injury group(P<0.05).Moreover,significantly increased Bcl-2 expression and decreased Bax expression were observed in the combined intervention group as compared with the cell injury group.In conclusion,ginsenoside Rg1 can antagonize hydrogen peroxide-induced damage to rat bone marrow-derived endothelial progenitor cells,increase cell viability,inhibit cell apoptosis,and reduce oxidative stress injury via the Erk signaling pathway.
作者
蒋文捷
梁雪梅
Jiang Wen-jie;Liang Xue-mei(Department of Gerontology,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第33期5338-5343,共6页
Chinese Journal of Tissue Engineering Research
作者简介
蒋文捷,男,1987年生,2015年西南医科大学毕业,硕士,医师,主要从事老年病学方面的研究;通讯作者:梁雪梅,博士,副教授,副主任医师,西南医科大学附属医院老年科,四川省泸州市646000
