阿魏酸川芎醇酯对氧化损伤内皮细胞ICAM-1表达的影响

摘要

Objective To investigate the protective effect of 2-(4-hydroxy-3-methoxycinnamon acyloxymethyl)-3,5,6-trimethylpyrazine(TMPF) on oxidative damage in human umbilical vein endothelial cells (ECV304) , and to explore the mechanism of TMPF action. Methods The ECV304 cells were damaged by hydrogen peroxide and the protective effects of TMP and TMPF on oxidative damage were observed (Determination of MDA content and SOD vitality ac-cording to kit instructions s Determination of expression of ICAM-1 by real-time PCR: Determination of the expression of ICAM-1 protein by Elisa). Results Compared with normal control cells, hydrogen peroxide induced an increased in malondialdehyde (MDA) content and a decrease in su-peroxide dismutase (SOD) activity (P<0. 05 or P<0. 01). However,TMP or TMPF treatment sifnificantly inhibited hydrogen peroxide-induced MDA formation and increased SOD activity (P<0. 05 or P<0. 01). Compared TMP treatment,TMPF treatment reduced MDA formation and elevated SOD activity. But no significant reduction in SOD activity was obtained by treatment with TMP or TMPF at dose of 12. 5 μmol o L-1 (P>0. 05). The expression of ICAM-1 mRNA in normal control group,25. 0 fimol o L-1TMP treatment group and 25. 0 μmol o L-1 TMPF treatment group was 0. 245,0. 454 and 0. 376 times higher than that in hydrogen peroxide treatment group,respectively. Compared with hydrogen peroxide treatment group,the expression of ICAM-1 protein significantly decreased in normal control group,25. 0 μmol o L-1TMP treatment group and 25. 0 μmol o L-1 TMPF treatment group(P<0. 05 orP<0. 01). Conclusion TMPF can protect hydrogen peroxide-induced oxidative damage, decrease MDA formation and increased SOD activity. TMPF may reduce ICAM-1 expression to attenuate oxidative damage in endothelial cells.%目的 观察川芎嗪衍生物阿魏酸川芎醇酯(TMPF)对H2O2引起的脐静脉内皮细胞(ECV304)氧化性损伤的保护作用,并初步探讨其作用机制.方法 以H2O2损伤ECV304细胞建立氧化损伤模型,以川芎嗪(TMP)作为阳性对照药物,观察TMPF对氧化损伤的ECV304细胞的保护作用[按试剂盒说明测量微量丙二醛(MDA)含量及超氧化物歧化酶(SOD)活力；采用Real-Time PCR测定ICAM-1 mRNA含量；采用Elisa检测ICAM-1蛋白表达].结果 与正常对照组比较,H2O2损伤组细胞内的MDA含量明显升高(P＜0.01),TMP与TMPF各浓度保护组MDA含量明显低于H2O2损伤组(P＜0.05或P＜0.01),且TMPF保护组MDA含量均低于同剂量TMP保护组(P＜0.05)；与正常对照组比较,H2O2损伤组细胞内的SOD水平明显降低(P＜0.05或P＜0.01),在12.5 μmol·L-1 TMP保护组和TMPF保护组与H2O2损伤组相比差异无统计学意义(P＞0.05),在TMP和TMPF的其他浓度上与H2O2损伤组相比差异均有统计学意义(P＜0.05或P＜0.01).H2O2可上调ICAM-1 mRNA的含量,正常对照组ICAM-1 mRNA的含量为H2O2损伤组的0.245倍,25.0μmol·L-1TMP保护组ICAM-1 mRNA的含量为H2O2损伤组的0.454倍,25.0 μmol· L-1 TMPF保护组ICAM-1 mRNA的含量为H2O2损伤组的0.376倍.H2O2损伤组细胞ICAM-1蛋白的表达量显著高于正常对照组(P＜0.01),25.0 μmol·L-1TMP 保护组ICAM-1蛋白的表达量低于H2O2损伤组(P＜0.05),而25.0 μmol·L-1 TMPF保护组ICAM-1蛋白的表达量显著低于H2O2损伤组(P＜0.01).结论 TMPF对H2O2引起内皮细胞氧化性损伤有明显的保护作用,能减少内皮细胞脂质过氧化物MDA的生成,提高SOD活性,TMPF能降低损伤细胞ICAM-1 mRNA的含量及其蛋白表达.TMPF对氧化损伤内皮细胞中ICAM-1表达上调有较强的抑制作用,这可能是TMPF对内皮细胞氧化损伤保护作用的重要机制.