【摘要】 目的观察血管钙化大鼠模型主动脉醛固酮及其受体表达的变化。方法 21只7周龄雄性SD大鼠随机分为正常组、钙化组、钙化+螺内酯组,每组7只。钙化组和螺内酯组采用维生素D3(300000U/kg一次肌肉注射)和尼古丁(25mg/kg溶于花生油中早、晚各灌胃1次)诱导大鼠血管钙化模型。螺内酯组造模第二天给予螺内酯40mg/(kg·d)灌胃,持续6周。用钙离子测试盒、碱性磷酸酶试剂盒测定钙含量和碱性磷酸酶活性,VonKossa染色检测血管钙化程度,放射免疫法检测大鼠血浆和血管组织的醛固酮含量,免疫组织化学法检测血管组织盐皮质激素受体表达。结果 VonKossa染色可见血管钙化大鼠主动脉有大量黑色颗粒沉淀,钙化组的血管钙含量、碱性磷酸酶活性明显高于正常组[分别为(0.40±0.05)比(0.23±0.03)μmol/g蛋白,(188.00±31.37)比(112.00±20.10)U/g蛋白,均P<0.01]。钙化组的血管醛固酮含量及其受体表达比正常组明显上调。与钙化组相比,使用螺内酯能够减轻血管钙含量、碱性磷酸酶活性和减少血管醛固酮含量[(0.28±0.04)比(0.40±0.05)μmol/g蛋白,(143.90±26.17)比(188.00±31.37)μmol/g蛋白,(21.80±3.54)比(28.43±4.47)ng/g蛋白,均P<0.05]。3组血浆醛固酮水平差异无统计学意义(P>0.05)。结论钙化大鼠血管组织醛固酮及其受体表达上调,提示醛固酮可能参与了血管钙化的发生和发展过程。更多还原

【Abstract】 Objective To investigate the expressions of aldosterone and its receptor on aorta in a rat vascular calcification model. Methods Twenty-one seven-week-old male SD rats were randomly divided into normal , calcification and calcification plus spironolactone group ( n=7 / group ) . On the first day in calcification and spironolactone group , vascular calcification was induced by a single dose intramuscular injection of vitamin D3 ( 300 000U / kg ) in the morning plus two doses of nicotine ( 25 mg / kg dissolved in peanut oil ) gavage in the morning and in evening. The next day after calcification induction , rats in spironolactone group were gavaged with spironolactone [ 40mg /( kg·d )] for 6weeks. Calcium content and alkaline phosphatases ( ALP ) activity were detected using calcium assay kit and alkaline phosphatase detection kit respectively. Vascular calcification was determined by Von Kossa staining. Aldosterone contents in plasma and aorta were determined by radioimmunoassay , and mineralocorticoid receptor immuneactivity was determined by immunohistochemistry. Results Von Kossa staining showed that mass black granules deposited in aortic wall of the vascular calcified rats. Calcium content and ALP activity in calcified group were increased significantly than those in the control group [( 0.40±0.05 ) vs ( 0.23±0.03 ) μ mol / g ( protein ), ( 188.00±31.37 ) vs ( 112.00±20.10 ) U / g ( protein ), both P<0.01 ] . Meanwhile , aortic aldosterone [( 28.43±4.47 ) vs ( 15.07±5.00 ) ng / g ( protein ), P<0.01 ] and mineralocorticoid receptor level in calcified group were up-regulated significantly compared with those in the control group. In addition , spirolactone diet could reduce the degree of vascular calcium contents , ALP activity and aldosterone content [( 0.28±0.04 ) vs ( 0.40±0.05 ) μ mol / g ( protein ),( 143.90±26.17 ) vs ( 188.00±31.37 ) μ mol / g ( protein ),( 21.80±3.54 ) vs ( 28.43±4.47 ) ng / g ( protein ), all P< 0.05 ] . There were no significant differences in plasma aldosterone contents among these groups ( P>0.05 ) . Conclusion Aldosterone and mineralocorticoid receptor were significantly increased in calcified vessels suggesting that aldosterone may be involved in the development of vascular calcification.更多还原