摘要
目的探讨血管紧张素Ⅱ(AngⅡ)对肺微血管内皮细胞APJ mRNA和Apelin mRNA表达的影响。方法培养并鉴定24h新生SD大鼠肺微血管内皮细胞,取2-4代培养细胞,细胞密度5×10^5个/ml,培养至80%融合后,进行药物干预。第一部分实验分为5组(n=4):加入AngⅡ至终浓度分别为10^-9mol/L(Ⅱ组)、10^-8mol/L(Ⅲ组)、10^-7mol/L(Ⅳ组)、10^-6mol/L(Ⅴ组),Ⅰ组不加入AngⅡ,24h后采用RT-PCR技术测定Apelin mRNA和APJ mRNA的表达水平。第二部分实验分为6组(n=4):加入AngⅡ至终浓度为10^-7mol/L,分别在孵育即刻(Ⅰ组)、1h(Ⅱ组)、6h(Ⅲ组)、12h(Ⅳ组)、24h(Ⅴ组)、48h(Ⅵ组)时,采用RT-PCR技术测定Apelin mRNA和APJ mRNA的表达水平。结果第一部分实验结果:与Ⅰ组比较,Ⅱ组Apelin mRNA表达上调,APJ mRNA表达下调,Ⅲ组~Ⅴ组Apelin mRNA和APJ mRNA表达下调(P〈0.05或0.01),与Ⅱ组比较,Ⅲ组~Ⅴ组Apelin mRNA和APJ mRNA表达下调,呈浓度依赖性(P〈0.01)。第二部分实验结果:Apelin mRNA在10^-7mol/L AngⅡ孵育6h内表达上调,孵育1h时达高峰(P〈0.05或0.01),孵育6h后Apelin mRNA表达下调,且呈时间依赖性(P〈0.01);而APJ mRNA在孵育12h后呈时间依赖性持续下调(P〈0.01)。结论AngⅡ呈浓度和时间依赖性地下调Apelin mRNA和APJ mRNA的表达,可能是其参与肺微血管内皮细胞损伤的发生机制。
Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ ) on the Apelin mRNA and APJ mRNA expression in pulmonary microvascular endothelial cells (PMVECs) in rats. Methods Pulmonary microvascular endothelial cells obtained from 24 h old neonatal SD rats were cultured in DMEM liquid culture medium. The 2nd-4th generation PMVECs were inoculated on 6-well plates (5 × 10^5 ). The experiment was performed in two parts. In part Ⅰ different concentration of AngⅡ 0, 10^-9 , 10^-8 , 10^-7 , 10^-6mol/L (group Ⅰ - Ⅴ ) were added into the PMVECs. The expression of Apelin mRNA and APJ mRNA was determined at 24 h after addition of AngⅡ by RT-PCR. In part Ⅱ the cells were exposed to 10^-7mol/L AngⅡ . The expression of Apelin mRNA and APJ mRNA was determined immediately and at 1, 6, 12, 24, 48 h(group Ⅰ -Ⅵ ) after addition of Ang Ⅱ by RT-PCR. Results In part Ⅰ Apelin mRNA expression was significantly higher in group Ⅱ (Ang Ⅱ 10^-9 mol/ L) but lower in group Ⅲ-Ⅴ (AngⅡ 10^-8 s, 10^-7, 10^-6 mol/L) than in group Ⅰ (control, AngⅡ 0 mol/L). The APJ mRNA expression was significandy lowered in group Ⅱ -Ⅴ in a dose-dependent manner as compared with control group (group Ⅰ ) . In part Ⅱ both Apelin mRNA and APJ mRNA expression exhibited a bi-phasic response to Ang Ⅱ 10^-7 mol/L, increased at first and was then decreasing with time. The Apelin mRNA and APJ mRNA expression reached the peak at 1 h of incubation with Ang Ⅱ respectively. Conclusion Ang Ⅱ decreases both Apelin mRNA and APJ mRNA expression in PMVECs in a dose and time dependent manner. The down-regulation of Apelin mRNA and APJ mRNA expression may be involved in the mechanism of injury to PMVECs.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2008年第9期824-827,共4页
Chinese Journal of Anesthesiology
关键词
受体
血管紧张素
2型
内皮细胞
血管
肺
受体
G-蛋白偶联
配体
Receptor, angiotensin, type 2
Endothelial cells
Blood vessels
Lung
Receptors, G-protein-coupled
Ligands