黄芪总黄酮对尿毒症患者血清诱导的内皮细胞凋亡的影响

摘要

目的：探讨黄芪总黄酮（T FA ）对尿毒症患者血清诱导的内皮细胞凋亡的影响。方法收集22例健康志愿者和25例规律血液透析的尿毒症患者血清。以人脐静脉血管内皮细胞（HUVECs）为研究对象，设立对照组（细胞同步化后加入健康志愿者血清）和尿毒症组（细胞同步化后加入尿毒症患者血清）。细胞同步化前6h，在尿毒症组的一部分细胞中加入0．5、1．0、2．0 mg/mL T FA预先进行干预，分别得到低剂量组、中剂量组、高剂量组细胞。细胞培养24 h后，于显微镜下观察细胞形态；M T T法检测细胞增殖活力；黄嘌呤氧化酶法测定超氧化物歧化酶（SOD）活性；硝酸还原酶比色法测定一氧化氮（NO）水平；彗星实验法检测细胞DNA损伤；TUNEL法检测细胞凋亡。结果与对照组比较，尿毒症组细胞增殖活力、SOD活性、NO水平均降低（P＜0．01），DNA拖尾率、细胞凋亡指数（AI）均升高（P＜0．01）；与尿毒症组细胞比较，不同剂量干预的各组细胞增殖活力均增加（P＜0．05），NO水平均升高（P＜0．01）；与尿毒症组比较，中剂量、高剂量组SOD活性增加（P＜0．05），DNA 损伤拖尾率降低（P＜0．05）。结论 TFA可减少尿毒症患者血清诱导的内皮细胞凋亡，其可能机制与抗氧化应激有关。%Objective To explore the effect of total flavonoids of astragalus (TFA) on the apoptosis of endothelial cells induced by serum of uremia patient .Methods The serum of 22 healthy volunteers and 25 uremia patients receiving regularly hemodialysis were enrolled in the study .HUVECs were used as research objects ,which were divided into control group(adding serum of healthy people when cell synchronized) and uremia group (adding serum of uremia patient when cell synchronized ) .Low dose ,moderate dose and high dose group were prepared by adding 0 .5 ,1 .0 ,2 .0 mg/mL TFA respectively 6 h before cell synchronization .After 24 hours′culture since the serum were added ,the morphological change of endothelial cells were observed by microscopy ,proliferation activities were tested by using MTT ,SOD activities were tested by using xanthine oxidase method ,NO levels were measured by u-sing nitrate reductase colorimetric method ,DNA damage was detected by using comet assay ,the morphological change of apoptosis was observed by using TUNEL method .Results Compared with the control group ,the proliferation activity ,SOD activity ,NO lev-els were lower in uremia group(P< 0 .01) ,DNA tailing rate ,apoptosis index(AI) significantly increased (P<0 .01) .Compared with cells of uremia group ,cell proliferation activity of all the TFA intervention groups increased (P<0 .05) ,NO levels also in-creased (P<0 .01) .Compared with uremia group ,moderate and high dose group′s SOD activity increased (P<0 .05) ,DNA damage tailing rate decreased (P<0 .05) .Conclusion Total flavonoids of astragalus reduces apoptosis of HUVECs induced by serum of uremia patient ,the possible mechanism is associated with the decrease of oxidative stress .