血管紧张素Ⅱ对大鼠肺微血管内皮细胞APJ mRNA和Apelin mRNA表达的影响

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24 h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5×105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10-9 mol/L(Ⅱ组)、10-1 mol/L(Ⅲ组)、10-7mol/L(Ⅳ组)、10-6 mol/L(Ⅴ组),Ⅰ组不加入AngⅡ,24 h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)、1 h(Ⅱ组)、6 h(Ⅲ组)、12 h(Ⅳ组)、24 h(Ⅴ组)、48 h(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6 h内表达上调,孵育1 h时达高峰(P<0.05或0.01),孵育6 h后Apelin mRNA表达下调,且呈时间依赖性(P<0.01);而APJ mRNA在孵育12 h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.

Effect of angiotension Ⅱ on Apelin mRNA and APJ mRNA expression in pulmonary microvascular endothelial cells in rats

Objective To investigate the effects of angiotensinⅡ (AngⅡ) on the Apelin mRNA and APJ mRNA expression in pulmonary microvascular endothelial ceils (PMVECs) in rats. Methods Pulmonary microvascular endothelial ceils 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 (5×105). 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 Ang Ⅱ by RT-PCR. In part Ⅱ the cells were exposed to 10-7 mol/L Ang Ⅱ. 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, 10-7, 10-6 mol/L) than in group Ⅰ (control, Ang Ⅱ 0 mol/L). The APJ mRNA expression was significantly lowered in group Ⅱ-Ⅴ in a dose-dependent manner as compared with control group (group Ⅰ). In partⅡ beth Apelin mRNA and APJ mRNA expression exhibited a bi-phasic response to AngⅡ 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.