高血压合并冠心病患者平滑肌细胞内Ca2+水平和血管紧张素Ⅱ1型受体表达的观察研究

目的 观察高血压合并冠心病患者与正常血压冠心病患者平滑肌细胞内Ca2+水平和血管紧张素Ⅱ1型(AT1)受体的表达情况.方法 ①收集首都医科大学附属北京安贞医院心脏外科行冠状动脉旁路移植术患者术中剩余大隐静脉,进行细胞培养,分为高血压搭桥组和正常血压搭桥组.②用激光共聚焦显微镜(CLSM)观察不同浓度的血管紧张素Ⅱ(AngⅡ)分别刺激后2组的平滑肌细胞内游离钙浓度的变化情况.③提取培养的平滑肌细胞总RNA、RT-PCR,观察2组AT1受体的表达情况.结果 经AngⅡ刺激后人平滑肌细胞内Ca2+均迅速升高.在高血压搭桥组AngⅡ刺激后平滑肌细胞胞内Ca2+荧光吸光度较正常血压搭桥组明显升高(P＜0.05).在平滑肌细胞观察到高血压搭桥组AT1受体表达较正常血压搭桥组略高,差异有统计学意义(P＜0.05).结论 AngⅡ刺激后人血管平滑肌细胞存在胞内Ca2+的变化.高血压冠心病患者和正常血压冠心病患者平滑肌细胞存在AT1受体表达差别.

Study on the expression of free Ca2+ and AngiotensinⅡ1 type receptor in vascular smooth muscle cells of hypertensive coronary artery diseases

Objective To observe the expression of free Ca2+ and Angiotensin Ⅱ 1 type receptor(AT1 R)in vascular smooth muscle cells(VSMCs)of patients with hypertensive or normotensive coronary artery diseases(CAD).Methods During the coronary artery bypass graft operation,the surplus saphenous vein of patients who admitted to our Cardiac Surgery Department was collected and cultured.All patients were divided into hypertensive or normotensive group.Free Ca2+ in the cultured human VSMCs was determined by confocal laser scanning microscope(CLSM)after different kinds of AngiotensinⅡ being added,respectively.Total RNA was extracted from cultured VSMCs.Then RT-PCR was conducted for the observation of the expression of AT1R in both groups.Results Ca2+in human VSMCs rapidly increased when stimulated by Angtensin Ⅱ in two groups.After stimulated by Angiotensin Ⅱ,both free Ca2+ level and the expression of AT1R in VSMCs of hypertensive patients were higher than those of the normotensive patients(P＜0.05).Conclusion There are certain changes of free calcium in the cultured human vascular smooth muscle cells,when stimulated by Angiotensin Ⅱ.There are also difierences in AT1R expression between hypertensive CAD patients and normotensive CAD patients.