黄芪甲苷对过氧化氢诱导损伤的人脐静脉内皮细胞的保护作用

马海涛,王　辉

1)河南中医学院第一附属医院周围血管科 郑州 450000　2)河南中医学院药学院中药教研室 郑州 450000

△男,1969年6月生，本科，副主任医师，研究方向：中西医结合治疗周围血管疾病，E-mail：mahaitao6968@126.com



黄芪甲苷对过氧化氢诱导损伤的人脐静脉内皮细胞的保护作用

马海涛1)△,王辉2)

1)河南中医学院第一附属医院周围血管科 郑州 4500002)河南中医学院药学院中药教研室 郑州 450000

△男,1969年6月生，本科，副主任医师，研究方向：中西医结合治疗周围血管疾病，E-mail：mahaitao6968@126.com

关键词下肢缺血；血管内皮细胞；DNA损伤；黄芪甲苷；Wnt/β-catenin信号通路

摘要目的：探讨黄芪甲苷对过氧化氢诱导损伤的人脐静脉内皮细胞(HUVECs)的保护作用，并初步探讨其作用机制。方法：实验Ⅰ中取HUVECs后分为5组，其中4组分别给予不同质量浓度的黄芪甲苷预处理后，用过氧化氢诱导建立HUVECs损伤模型，对照组不处理。实验Ⅱ中另取HUVECs分为4组，即对照组、LiCl组、黄芪甲苷组及LiCl+黄芪甲苷组，后3组按分组处理后均同上用过氧化氢诱导。采用MTT法检测上述各组细胞存活情况，流式细胞术检测细胞凋亡，Western blot分析γ-H2AX、β-catenin蛋白的表达。结果：过氧化氢可抑制HUVECs存活，诱导其凋亡，而黄芪甲苷可改善上述情况(P<0.05)。Wnt通路激动剂LiCl预处理后，黄芪甲苷对损伤HUVECs的保护作用减弱，同时DNA损伤标记分子γ-H2AX以及β-catenin蛋白的表达水平升高(P<0.05)。结论：黄芪甲苷可缓解氧化应激诱导的HUVECs损伤，其机制可能与抑制Wnt/β-catenin信号通路有关。

Protective effects of astragaloside on injured human umbilical vein endothelial cells induced by H2O2

MAHaitao1),WANGHui2)

1)DepartmentofPeripheralVascular,theFirstAffiliatedHospital，HenanUniversityofTraditionalChineseMedicine,Zhengzhou4500002)DepartmentofTraditionalChineseMedicine，CollegeofPharmacy,HenanUniversityofTraditionalChineseMedicine,Zhengzhou450000

Key wordshindlimb ischemia;vascular endothelial cell;DNA injury;astragaloside;Wnt/β-catenin signaling pathway

AbstractAim: To study the effect and the underlying mechanism of astragaloside on injured human umbilical vein endothelial cells(HUVECs) induced by H2O2. Methods: In experiment Ⅰ, HUVECs were allocated into 5 groups,among which, 4 groups were pretreated by astragaloside at different doses and then treated by H2O2，and the control group was not treated. In experiment Ⅱ, HUVECs were allocated into 4 groups as control group, LiCl group, astragaloside group, and LiCl+astragaloside group and treated accordingly,and then they were treated by H2O2，and the control group was not treated.MTT assay was used to assess the survival of HUVECs, and cell apoptosis was analyzed by flow cytometry. Furthermore, the expression levels of γ-H2AX and β-catenin protein were detected by Western blot. Results: Astragaloside attenuated the inhibitory effect of H2O2on cell survival and down-regulated the apoptosis rate of HUVECs induced by H2O2. Further mechanism analysis showed that pretreatment with LiCl attenuated the protecting effect of astragaloside on HUVECs with an obvious increase in the expression levels of γ-H2AX and β-catenin protein(P<0.05). Conclusion: Astragaloside could protect the HUVECs from oxidative stress mainly by regulating the Wnt/β-catenin signaling pathway.

下肢缺血性疾病在周围血管疾病(peripheral artery disease,PAD)中最为常见，部分甚至可发展成严重肢体缺血，患者最后不得不承受截肢的痛苦，严重影响患者的生活质量[1]。目前临床上治疗下肢缺血的方法主要有药物治疗和手术治疗等，但效果并不理想。研究[2]证实，血管阻塞后大量的氧自由基诱发的内皮细胞DNA损伤是影响下肢缺血病理进程的重要因素。……