特女贞苷对血管内皮细胞氧化损伤的作用研究  

Effect of Specnuezhenide Reducing the Oxidative Damage of Vascular Endothelial Cells

作　　者：顾闻[1] 刘特[1] 陈久林[1] 申定珠[1] 邢三丽[1] 郁志华[1] 陈川[1] GU Wen;LIU Te;CHEN Jiu-lin;SHEN Ding-zhu;XING San-li;YU Zhi-hua;CHEN Chuan(Shang hai Geriatric Institute of Chinese Medicine,Shanghai（200031)

机构地区：[1]上海市中医老年医学研究所,上海200031

出　　处：《中国中西医结合杂志》2018年第9期1093-1098,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81373706;No.81202731;No.81202811)

摘　　要：目的研究特女贞苷对过氧化氢诱导的人脐静脉内皮细胞(EA.hy926)中p66Shc及凋亡相关蛋白表达水平的作用。方法采用H_2O_2诱导EA.hy926形成氧化应激损伤的血管内皮细胞模型,随机分为空白组、模型组(H_2O_2)、特女贞苷组(H_2O_2+5、50、500μmol/L特女贞苷)、阴性对照组(H_2O_2+5、50、500μmol/L DMSO)、抗坏血酸组(H_2O_2+5、50、500μmol/L抗坏血酸)。检测各组细胞内超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量、β-gal活性;采用Western blot法测定p66Shc、caspase-3活体剪切体、Bax、Bcl-2蛋白含量;实时荧光定量PCR(qPCR)法检测p66mRNA含量。结果与空白组比较,模型组及阴性对照组SOD活性、Bcl-2表达量及Bcl-2/Bax比值下降,MDA含量、β-gal活性、p66Shc蛋白水平及mRNA含量、capase-3活性剪切体和Bax表达水平升高(均P<0.05)。与模型组比较,特女贞苷组及抗坏血酸组SOD升高,MDA含量(除特女贞苷组浓度500μmol/L外)和β-gal活性降低(P<0.01),抑制p66Shc表达及mRNA水平(P<0.01);特女贞苷组Bax和capase-3活性剪切体(除浓度500μmol/L外)表达水平降低,Bcl-2表达和Bcl-2/Bax比值升高(均P<0.01)。与同浓度的抗坏血酸组比较,特女贞苷组对Bax、Bcl-2蛋白水平、Bcl-2/Bax比率、p66mRNA的调节作用较明显(P<0.05)。结论特女贞苷可能通过抑制p66Shc削弱血管内皮细胞的氧化损伤及降低凋亡相关蛋白的表达。Objective To observe the effect of Specnuezhenide on the expressions of p66Shc and apoptosis-related protein of hydrogen peroxide-induced human umbilical vein endothelial cells （EA. hy926）. Methods Oxidative damage model of vascular endothelial cells was established H2O2 induction on EA.hy926. EA.hy926 cells were divided into the blank group, the model group （H2O2）, the specnu- ezhenide group （H2O2 ＋5,50,500μmol/L of specnuezhenide）, the negative control group （H2O2 ＋5,50, 500 μmol/L of DMSO）, and the ascorbic acid group （H2O2 ＋ 5, 50, 500μmol/L of ascorbic acid）. Levels of the superoxide dismutase （SOD） activity, malonic dialdehyde （MDA） content, and 13-gal activity of each group were detected. Protein expression of p66Shc, cleaved caspase-3, Bax, Bcl-2 were detected by Western blot. mRNA expressions of p66 were detected by real-time fluorescent quantitative PCR （qPCR）. Results Compared with the blank group, SOD activity, Bal-2/Bax ratio and the expression of Bcl-2 decreased, while MDA content, β-gal activity, the expression levels of p66Shc protein, p66 mR- NA, cleaved caspase-3, Bax increased in the model group and the negative control group （all P 〈0.05）. Compared with the model group, SOD activity increased,MDA content （except for the concentration of 500 μmol/L in the specnuezhenide group） and β-gal activity decreased, the expression of p66Shc and mRNA were inhibited in the specnuezhenide group and the ascorbic acid group （all P 〈0.01 ）. The expres- sions of bax and cleaved capase-3 （except concentration 500 μmol/L） were decreased, the expressions of Bcl-2 and Bcl-2/Bax ratio were increased （all P 〈0.01 ） in the specnuezhenide group. Compared withthe same concentration of the ascorbic acid group, the specnuezhenide group had a significant effect on the regulation of Bax, Bcl-2 protein levels, Bcl-2/Bax ratio, and p66 mRNA （P 〈0.05）. Conclusion Specnuezhenide cloud reduce the oxidative damage and the expression of apoptosis-related

关 键 词：P66SHC 氧化应激 特女贞苷 人脐静脉内皮细胞 

分 类 号：R285[医药卫生—中药学]