大鼠低氧性肺动脉高压肺血管重构模型的构建及肺组织中miR-34a的表达  

Construction of Pulmonary Vascular Remodeling Model of Hypoxic Pulmonary Hypertension in Rats and the Expression of MicroRNA-34a in Lung Tissue

作　　者：马兰[1,2] 刘川川 周振[1,2] 宋泽青 刘洁[1] 格日力 MA Lan1＇2, LIU Chuan-chuan1＇2, ZHOU Zhen1＇2, SONG Ze-qing3, LIU JieI, GE Ri-li 1,2(1.Research Center for High Altitude Medicine, Qinghai University Medical College, Xining, Qinghai 810001 ; 2.The Key Laboratory of High Altitude Medical Application of Qinghai Province,Xining,Qinghai 810001; 3.Department of Clinical Medicine,Qinghai University Medical College,Xining,Qinghai 810001)

机构地区：[1]青海大学医学院高原医学研究中心,青海西宁810001 [2]青海省高原医学应用基础重点实验室,青海西宁810001 [3]青海大学医学院临床医学系,青海西宁810001

出　　处：《中国高原医学与生物学杂志》2018年第1期6-12,共7页Journal of Chinese High Altitude Medicine & Biology

基　　金：国家自然科学基金项目(No.31560292);青海省科技厅自然科学青年基金项目(2015-ZJ-939Q);青海大学医学院中青年基金项目(2014-KY-1)

摘　　要：目的构建大鼠低氧性肺动脉高压肺血管重构模型,观察P53基因及miR-34a在模型大鼠肺组织中的表达情况。方法 40只雄性SD大鼠随机分为常氧对照组和低氧1周(1wk)组、低氧2周(2wk)组及低氧3周(3wk)组,各组10只。低氧组大鼠置大型低压氧舱(模拟海拔5000m)构建低氧性肺动脉高压肺血管重构模型,正常对照组在低压氧舱外正常饲养。用生理信号采集系统记录肺动脉压,并计算右心室质量比来评定判断模型是否构建成功。用Real-time PCR法测定大鼠肺组织中miR-34a和P53 mRNA的表达情况、western blot法测定大鼠肺组织中P53的蛋白表达情况,并行相关分析。结果在低氧处理的第1、2、3wk RV/(LV+S)及mPAP明显增加,差异有统计学意义(P<0.01)。大鼠肺组织中P53基因表达明显增高,呈时间依赖性;miR-34a的表达也随着低氧时间的延长逐渐增高,在第3周达最高,差异有统计学意义(P<0.01)。结论成功构建了低氧性肺动脉高压肺血管重建模型;P53基因和miR-34a随着低压低氧时间的延长表达上调。Objective To construct pulmonary vascular remodeling with hypoxic pulmonary hypertension model in rats, and to observe the expression of P53 gene and miR-34a in the lung tissue of the rat model.Method 40 male SD rats were randomly divided into normoxic control group and hypoxic 1 week group, hypoxic 2 weeks group andhypoxic 3 weeks group, 10 rats in each group.And hypoxic groups were kept in a hypobaric chamber stimulated the altitude of 5 000 m with hypoxia time.The normoxic control group were raised outside of hypobaric chamber. The physiological signal acquisition system was used to record pulmonary artery pressure, and RV/（ LV ＋S）ratio were Calculated, and to assess whether the model is successful.The expression of miR-34a and P53 mRNA in lung tissue of rats were measured by Real-time PCR method,and the protein expression of P53 in lung tissue of rats were measured by western blot.Results The mPAP and RV/（ LV＋S）ratio were significantly increased in hypoxic 1 week group, hypoxic 2 weeks group and hypoxic 3 weeks group.It indicated that the hypoxic pulmonary hypertension ani- mal models were successfully constructed with prolonged hypoxia in a hypobaric chamber. The expression of P53 gene in rat lung tissue increased significantly with time dependent.And the expression of miR-34a increased with the prolongation of hypoxia time, and reached the highest level at the third hypoxic weeks, the difference was statisti- cally significant.Conclusion We successfully constructed pulmonary vascular remodeling model of hypoxic pulmona- ry hypertension in rats.And the expression of P53 gene and miR-34a were up-regulated with the prolongation of low pressure and hypoxia time.

关 键 词：低氧 肺血管重建 肺动脉高压 microRNA-34a 

分 类 号：R363[医药卫生—病理学]