吉马酮改善H2O2诱导的脐静脉血管内皮细胞氧化应激损伤的作用(1)
[摘要]該文主要研究吉马酮对过氧化氢(H2O2)诱导的脐静脉血管内皮细胞(HUVECs)氧化损伤的保护作用并探讨其可能的作用机制。使用500 μmol·L-1H2O2诱导脐静脉内皮细胞3 h，从而建立氧化损伤模型，再用不同浓度吉马酮(20，40，100，150，200 μmol·L-1)保护24 h。利用MTT法检测吉马酮对H2O2损伤HUVECs细胞活力的影响；ELISA法检测PGI2，TXB2，ET1，tPA，PAI1，TNFα和IL6；硝酸还原酶法检测NO；比色法检测NOS及GSHPx；再分别采用TBA法、WST1法和微量酶标法检测MDA，SOD和LDH的含量等；Hoechst 33258荧光染色法观察细胞凋亡情况；RTPCR检测细胞中Bax，Bcl2，Caspase3 mRNA表达。结果表明500 μmol·L-1H2O2作用3 h细胞损伤率达到52%，而在20～200 μmol·L-1随着吉马酮浓度的增大被损伤细胞活性不断增强。与正常组比较，H2O2损伤使PGI2，NO，TNOS，tPA，SOD，GSHPx和Bcl2 mRNA降低，使PAI1，ET1，IL6，TNFα，TXB2，LDH，MDA，Bax mRNA和Caspase3 mRNA增加；与模型组比较，吉马酮(200，100，50 μmol·L-1)使PGI2，NO，TNOS，tPA，SOD，GSHPx和Bcl2 mRNA增加，使PAI1，ET1，IL6，TNFα，TXB2，LDH，MDA，Bax mRNA和Caspase3 mRNA减少。Hoechst 33258荧光染色与正常组比较，模型组细胞膜与细胞核呈浓缩致密的强蓝色荧光，细胞数量明显减少；与模型组比较，给药组的蓝色荧光强度降低等。以上研究结果表明，吉马酮可能通过抗氧化及抑制细胞凋亡等作用，从而改善H2O2诱导的脐静脉血管内皮细胞氧化应激损伤的作用。
, http://www.100md.com
[关键词]吉马酮； H2O2； 脐静脉内皮细胞； 氧化应激； 保护作用
Effect of germacrone in alleviating HUVECs damaged by
H2O2induced oxidative stress
CHEN Qiongfang1， WANG Gang1， TANG Liqing1， YU Xianwen1， LI Zhaofei2， YANG Xiufen1*
(1. Department of Pharmacology， School of Pharmacy， Guangxi University of Chinese Medicine， Nanning 530001， China；
2. Shaanxi Second People′s Hospital， Xi′an 710000， China)
, 百拇医药
[Abstract]This study focuses on the protective effect of germacrone on human umbilical vein endothelial cells(HUVECs) damaged by H2O2induced oxidative stress and its possible mechanisms. The oxidative damage model was established by using 500 μmol·L-1 H2O2 to treat HUVECs for 3 hours， and then protected with different concentrations of germacrone for 24 hours. The effect of germacrone on cell viability of HUVECs damaged by H2O2 was detected by MTT. The contents of PGI2， TXB2， ET1， tPA， PAI1， TNFα and IL6 were detected by ELISA. The content of NO was detected by using nitrate reductase method. Colorimetry was used to detect NOS and GSHPx. The contents of MDA， SOD and LDH were detected by TBA， WST1 and microplate respectively. Apoptosis was observed by Hoechst 33258 fluorescent staining. The mRNA expressions of Bax， Bcl2 and Caspase3 in cells were detected by RTPCR. The results showed that the cell damage rate was 52% after treated with 500 μmol·L1 H2O2 for 3 hours. The cell activity was increasing with the rise of germacrone concentration within the range of 20200 mol·L-1. Compared with normal group， the contents of PGI2， NO， TNOS， tPA， SOD， GSHPx and Bcl2 mRNA expressions were lower after damaged with H2O2. The contents of PAI1， ET1， IL6， TNFα， TXB2， LDH， MDA， Bax mRNA and Caspase3 mRNA expressions were increased. Compared with model group， the contents of PGI2， NO， TNOS， tPA， SOD， GSHPx and Bcl2 mRNA expressions were increased after treated with germacrone. The contents of PAI1， ET1， IL6， TNFα， TXB2， LDH， MDA， Bax mRNA and Caspase3 mRNA expressions were lower after treated with germacrone. According to Hoechst 33258 fluorescence staining， compared with normal group， the cell membrane and the nucleus showed strong dense blue fluorescence， and the number of cells significantly decreased in model group. Compared with model group， blue fluorescence intensity decreased in drug group. The above findings demonstrate that germacrone may improve the effect on HUVECs damaged by H2O2induced oxidative stress by resisting oxidation and inhibiting cell apoptosis., http://www.100md.com(陈琼芳王钢唐丽清 俞献文)
, http://www.100md.com
[关键词]吉马酮； H2O2； 脐静脉内皮细胞； 氧化应激； 保护作用
Effect of germacrone in alleviating HUVECs damaged by
H2O2induced oxidative stress
CHEN Qiongfang1， WANG Gang1， TANG Liqing1， YU Xianwen1， LI Zhaofei2， YANG Xiufen1*
(1. Department of Pharmacology， School of Pharmacy， Guangxi University of Chinese Medicine， Nanning 530001， China；
2. Shaanxi Second People′s Hospital， Xi′an 710000， China)
, 百拇医药
[Abstract]This study focuses on the protective effect of germacrone on human umbilical vein endothelial cells(HUVECs) damaged by H2O2induced oxidative stress and its possible mechanisms. The oxidative damage model was established by using 500 μmol·L-1 H2O2 to treat HUVECs for 3 hours， and then protected with different concentrations of germacrone for 24 hours. The effect of germacrone on cell viability of HUVECs damaged by H2O2 was detected by MTT. The contents of PGI2， TXB2， ET1， tPA， PAI1， TNFα and IL6 were detected by ELISA. The content of NO was detected by using nitrate reductase method. Colorimetry was used to detect NOS and GSHPx. The contents of MDA， SOD and LDH were detected by TBA， WST1 and microplate respectively. Apoptosis was observed by Hoechst 33258 fluorescent staining. The mRNA expressions of Bax， Bcl2 and Caspase3 in cells were detected by RTPCR. The results showed that the cell damage rate was 52% after treated with 500 μmol·L1 H2O2 for 3 hours. The cell activity was increasing with the rise of germacrone concentration within the range of 20200 mol·L-1. Compared with normal group， the contents of PGI2， NO， TNOS， tPA， SOD， GSHPx and Bcl2 mRNA expressions were lower after damaged with H2O2. The contents of PAI1， ET1， IL6， TNFα， TXB2， LDH， MDA， Bax mRNA and Caspase3 mRNA expressions were increased. Compared with model group， the contents of PGI2， NO， TNOS， tPA， SOD， GSHPx and Bcl2 mRNA expressions were increased after treated with germacrone. The contents of PAI1， ET1， IL6， TNFα， TXB2， LDH， MDA， Bax mRNA and Caspase3 mRNA expressions were lower after treated with germacrone. According to Hoechst 33258 fluorescence staining， compared with normal group， the cell membrane and the nucleus showed strong dense blue fluorescence， and the number of cells significantly decreased in model group. Compared with model group， blue fluorescence intensity decreased in drug group. The above findings demonstrate that germacrone may improve the effect on HUVECs damaged by H2O2induced oxidative stress by resisting oxidation and inhibiting cell apoptosis., http://www.100md.com(陈琼芳王钢唐丽清 俞献文)