论文部分内容阅读
目的:内皮素1(ET-1)和血管紧张素Ⅱ(AngⅡ)均能刺激肾上腺皮质细胞增殖,促进球状带细胞分泌醛固酮,但是,两者联合作用对皮质细胞产生的影响目前研究较少,涉及的信号传导通路尚不清楚。本实验拟研究ET-1和An-gⅡ联合处理对肾上腺皮质癌H295R细胞的ERK磷酸化的影响。方法:不同剂量ET-1和AngII刺激H295R细胞,用Western blot检测ERK1/2磷酸化水平;用AngⅡ 1型受体拮抗剂坎地沙坦及ETA拮抗剂BQ610预处理细胞,再加入ET-1及AngⅡ单独或共同刺激细胞,Western blot检测磷酸化ERK1/2。结果:ET-1和AngII均呈剂量依赖性刺激细胞ERK1/2磷酸化,两者的最佳刺激剂量均为100 nM;ETA受体拮抗剂BQ610能完全阻断ET-1的作用,而AngⅡ 1型受体拮抗剂坎地沙坦完全阻断AngⅡ的作用。ET-1和AngⅡ共处理细胞与两者单独刺激细胞在ERK1/2激活上无明显改变。结论:ET-1和AngII均激活H295R细胞ERK,且两者无交互作用。
PURPOSE: Endothelin-1 (ET-1) and angiotensinⅡ (AngⅡ) can stimulate the proliferation of adrenal cortical cells and promote the secretion of aldosterone by globular cells. However, there are few studies on the effect of the combination of both on the production of cortical cells. The signaling pathways involved are not clear. This study was to investigate the effect of ET-1 and An-gⅡ combined treatment on ERK phosphorylation in adrenocortical carcinoma H295R cells. Methods: The H295R cells were stimulated with different doses of ET-1 and AngII, and the phosphorylation of ERK1 / 2 was detected by Western blot. The cells were pretreated with canisartan, an antagonist of AngⅡ type 1 and BQ610, an ETA antagonist. ET-1 And Ang ¢ ò stimulated cells alone or jointly, and phosphorylated ERK1 / 2 was detected by Western blot. RESULTS: Both ET-1 and AngII phosphorylated ERK1 / 2 in a dose-dependent manner. The optimal stimulation dose was 100 nM. ETA receptor antagonist BQ610 completely blocked ET-1. AngⅡ Canisartan, a type 1 receptor antagonist, completely blocked Ang II. There was no significant change in ERK1 / 2 activation between ET-1 and AngⅡ co-treated cells and both stimulated cells alone. Conclusion: Both ET-1 and AngII activate ERK in H295R cells without interaction.