川芎内酯A预处理对心肌微血管内皮细胞缺氧/复氧损伤保护作用及机制研究

Protective effect and its mechanism of preconditioning of chuanxiong-pathalide A on rat cardiac microvascular endothelial cell injury by hypoxia and reoxygenation

目的:探讨川芎内酯A预处理对心脏微血管内皮细胞缺氧/复氧(H/R)损伤的保护作用及作用机制。方法:采用体外培养的大鼠乳鼠心脏微血管内皮细胞,空白组不加任何处理;单纯H/R组缺氧4 h/复氧2 h;单纯预处理组低氧25 m in/复氧30 m in,然后缺氧4 h/复氧2 h,各给药组分别加1×10-7,1×10-9,1×10-11mol.L-1的川芎内酯A孵育55 m in,然后缺氧4 h/复氧2 h;实验结束后取细胞上清液测一氧化氮(NO),一氧化氮合酶(NOS),内皮素(ET),取造模后的且固定在盖玻片的细胞,用原位杂交法检测iNOSmRNA和ETmRNA的基因表达。结果:H/R组细胞存活数降低明显,细胞培养液中NO和NOS活性显著降低,ET的活性明显升高,iNOSmRNA表达量减少,ETmRNA表达量增加;川芎内酯A 3个剂量组与H/R组比较,细胞存活数均明显增加,细胞培养液中NO和NOS活性增加,ET活性降低,iNOSmRNA表达量增加而ETmRNA表达量则减少。结论:川芎内酯A预处理能减轻内皮细胞损伤,对心脏微血管内皮细胞缺氧/复氧损伤可能具有保护作用,其作用机制可能与上调内皮细胞中iNOSmRNA的表达和下调ETmRNA的表达有关。

Objective： To investigate the protective effects and its mechanisms of chuanxiong-pathalide A on the injury of rat cardiac microvascular endothelial cells injury induced by the hypoxia and reoxygenation. Method： The cultured rat microvascular cells were incubated with chuanxiong-pathalide A in the concentrations of 1 × 10^-7 , 1 × 10^-9 , 1 × 10^-11 mol · L^-1 for 55 min, then suffered from hypoxia of 4 hours and reoxygenated of 2 hours. The activity of NO, NOS, ET and the expressions of iNOS mRNA and ET mRNA were observed. Result： Preconditioning with chuanxiong-pathalide A produced an increase in the survival rate of cells and the activitys of NO and NOS, and a decrease ET. Further, preconditioning with chuanxiong-pathalide A induced a high expression of iNOS mRNA and low expression of ET mRNA. Conclusion： The results showed that chuanxiong-pathalide A preconditioning can protect the cardiac microvascular endothelia from the injury caused by the hypoxia and reoxygenation, and the mechanism may be related to the up-regulation of iNOS mRNA expression and down-regulation of ET mRNA expression in the endothelium cell.