丹参酮ⅡA对猪主动脉内皮细胞受血管紧张素Ⅱ作用时产生NO及eNOS基因表达的影响

Effect of Tanshinone ⅡA on Angiotensin Ⅱ Induced Nitric Oxide Production and Endothelial Nitric Oxide Synthase Gene Expression in Cultured Porcine Aortic Endothelial Cells

目的探讨丹参酮ⅡA对血管内皮细胞的保护作用。方法采用硝酸还原酶法、逆转录聚合酶链式反应(RT-PCR)和免疫组织化学法,分别检测不同作用时间(1h、6h、24h)的血管紧张素Ⅱ(AngⅡ)以及在AngⅡ作用的不同时间点(0h点为A组、6h点为B组)加入丹参酮ⅡA对培养的猪主动脉内皮细胞产生NO及其内皮型一氧化氮合酶(eNOS)的蛋白和mRNA表达。结果(1)随着AngⅡ作用时间的延长,血管内皮细胞NO的产生及eNOS的表达呈显著下降(P<0.01),表现出时间依赖性的负性作用。(2)丹参酮ⅡA可抑制AngⅡ对内皮细胞分泌NO及eNOS基因表达的负性作用(P<0.01)。(3)在丹参酮ⅡA作用1h、6h,A组的抑制效应明显强于B组(P<0.05);随着作用时间延长至24h,两组比较差异无显著性。结论丹参酮ⅡA可抑制AngⅡ对血管内皮细胞分泌NO以及细胞eNOS基因表达的负性作用。

Objective To investigate the protective effect of tanshinone Ⅱ A on porcine aortic endothelial cells （PAEC）. Methods PAEC were stimulated with angiotensin Ⅱ （Ang- Ⅱ ） for different acting time （ 1 h, 6 h and 24 h） and Tanshinone Ⅱ A was added along with Ang- Ⅱ stimulation （Group A） or 6 h after it （Group B）. The nitric oxide （NO） level, the protein and mRNA expression of nitric oxide synthase （eNOS） in PAEC were measured by nitric acid deoxidizing assay, RT-PCR and immunohistochemical assay, respectively. Results With the prolongation of acting time of Ang- Ⅱ , the level of NO and eNOS expression in PAEC sequentially decreased in a negative acting time dependent manner （P 〈 0.01 ）, which could be inhibited by tanshinone Ⅱ A treatment independent to the dosage used （P 〈 0.01）. The inhibitory effect of tanshinone Ⅱ A was better in Group A than that in Group B either at 1 h or at 6 h after treatment （P 〈 0.05）. However, 24 h later, no significant difference was found between the effect in the two groups （P 〉 0.05）. Conclusion Tanshinone Ⅱ A could inhibit the negative effect of Ang- Ⅱ on NO production and eNOS expression in PAEC.