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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。
Objective To observe the changes of aldosterone and its receptor expression in the aortic vascular calcification rat model. Methods Twenty-one male Sprague-Dawley rats, 7 weeks old, were randomly divided into normal group, calcification group and calciferous + spironolactone group, with 7 rats in each group. Calcification group and spironolactone group using vitamin D3 (300000U / kg an intramuscular injection) and nicotine (25mg / kg soluble in peanut oil in the early and late each gavage once) induced vascular calcification model. Spironolactone 40mg / (kg · d) intragastrically on the second day of spironolactone group for 6 weeks. Calcium content and alkaline phosphatase activity were measured by calcium ion test kit and alkaline phosphatase kit. The degree of vascular calcification was detected by VonKossa staining. The content of aldosterone in plasma and vascular tissue was detected by radioimmunoassay. Mineralocorticoid receptor expression. Results VonKossa staining showed that a large number of black particles were precipitated in the aorta of rats with vascular calcification. The contents of calcium and alkaline phosphatase in calcification group were significantly higher than those in normal group [(0.40 ± 0.05) vs (0.23 ± 0.03) μmol / g Protein, (188.00 ± 31.37) vs (112.00 ± 20.10) U / g protein, all P <0.01]. Calcified group of vascular aldosterone content and its receptor expression was significantly higher than the normal group. Compared with the calcification group, spironolactone could reduce the content of calcium, alkaline phosphatase and decrease the content of aldosterone in blood vessel [(0.28 ± 0.04) vs (0.40 ± 0.05) μmol / g protein, (143.90 ± 26.17) vs 188.00 ± 31.37) μmol / g protein, (21.80 ± 3.54) vs (28.43 ± 4.47) ng / g protein, all P <0.05]. There was no significant difference in plasma aldosterone level between the three groups (P> 0.05). Conclusion The upregulation of aldosterone and its receptor in calcified rat blood vessels suggests that aldosterone may be involved in the occurrence and development of vascular calcification.