摘要
目的探讨葛根芩连汤(GQD)对高糖诱导的人脐静脉血管内皮细胞(HUVECs)氧化应激的影响。方法体外培养HUVECs。配置GQD中药复方含药血清,CCK-8法检测药物毒性、筛选实验最佳浓度及时间后将细胞分为对照组(细胞正常培养)、模型组(细胞利用60 mmol/L高糖处理48 h)、含药血清组(细胞先采用5%浓度含药血清预处理1 h后,再利用60 mmol/L高糖处理48 h)和阳性对照组[细胞先采用3 mmol/L N-乙酰半胱氨酸(NAC)预处理1 h后,再利用60 mmol/L高糖处理48 h]。细胞经以上处理后,采用CCK-8比色法检测细胞活力;流式细胞仪测试细胞凋亡及周期,同时检测细胞内活性氧(ROS)含量;生化试剂盒检测细胞肌酸激酶(CK)、谷胱甘肽(GSH)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)及丙二醛(MDA)水平。结果与对照组比较,模型组的细胞增殖率显著下降,细胞凋亡率、G0/G1期阻滞、ROS、CK、LDH及MDA的水平均显著增加,而SOD和GSH的水平显著下降(P<0.05)。与模型组比较,含药血清组和阳性对照组细胞的增殖率显著上升、凋亡率明显下降,同时G0/G1周期阻滞明显减弱(P<0.05),含药血清组和阳性对照组细胞的ROS、CK、LDH和MDA的水平均显著减少,GSH和SOD的水平明显上升(P<0.05)。结论葛根芩连汤可干预高糖诱导的血管内皮细胞氧化应激的发生,抑制细胞凋亡,提高细胞活力。
Objective To investigate the effect of Gegen Qinlian Decoction(GQD) on the oxidative stress of human umbilical vein vascular endothelial cells(HUVECs) induced by high glucose. Methods HUVECs were cultured in vitro. Chinese medicine compound GQD drug-containing serum were configured,CCK-8 method was used to detect drug toxicity,and to screen the optimal concentration and time of the experiment. The cells were divided into control group(normal cell culture),model group(cells treated with 60 mmol/L high sugar for 48 h),drug-containing serum group(cells firstly pretreated with 5% concentration drug-containing serum for 1 h, and then handled with 60 mmol/L hyperglycaemic treatment for 48 h)and positive control group [cells were pretreated with 3mmol/L N-acetylcysteine(NAC) for 1 h,and then treated with 60 mmol/L hyperglycemia for 48 h].After intervention,cell activity was detected by CCK-8 colorimetric method,cell apoptosis and cycle were detected by fow cytometry,and intracellular reactive oxygen(ROS)content was also detected. Biochemical kits were used to detect cell creatine kinase(CK),glutathione(GSH),lactate dehydrogenase(LDH),superoxide dismutase(SOD) and malondialdehyde(MDA).Results Compared with the control group,the cell proliferation rate of the model group decreased,the apoptosis rate,G0/G1 block,ROS,CK,LDH and MDA increased,while SOD and GSH levels decreased(P<0.05). Compared with the model group,the proliferation rate of cells in the drug-containing serum group and the positive control group increased,the apoptosis rate decreased,and the G0/G1 cycle block decreased(P<0.05). Compared with the model group,the content of ROS,CK,LDH and MDA in the drug-containing serum group and the positive control group decreased,and the levels of GSH and SOD increased(P<0.05). Conclusion GQD can interfere with the occurrence of oxidative stress of vascular endothelial cells induced by high glucose,inhibit apoptosis and improve cell vitality.
作者
陈俊
林晓
葛亚梅
钱紫星
朱梦杨
冉凤
CHEN Jun;LIN Xiao;GE Ya-mei;QIAN Zi-xing;ZHU Meng-yang;RAN Feng(Department of Endocrinology,Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan,430061;The First Clinical College,Hubei University of Traditional Chinese Medicine,Wuhan,430061;Clinical School of Traditional Chinese Medicine,Hubei University of Traditional Chinese Medicine,Wuhan,430061)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2023年第2期193-199,共7页
Chinese Journal of Integrated Traditional and Western Medicine
基金
湖北省卫生健康委员会中医药科研项目(面上项目)(No.ZY2021M016)。
关键词
葛根芩连汤
血管内皮细胞
增殖
凋亡
氧化应激
中药
Gegen Qinlian Decoction
vascular endothelial cells
proliferation
apoptosis
oxidative stress
Chinese herbal medicine
作者简介
通讯作者:陈俊,Tel:027-88920323,E-mail:chenjun1@hbhtcm.com。
