真武汤调控AVP/AQP2通路改善自发性高血压大鼠肾损伤研究

Zhenwu Decoction Regulates AVP/AQP2 Pathway to Improve Renal Injury in Spontaneously Hypertensive Rats

目的:探讨真武汤对自发性高血压大鼠(SHR)基于AVP/AQP2通路调控对肾损伤的保护作用。方法:50只SHR随机分为模型对照组、依那普利0.67 mg/kg组、真武汤2.1、8.4 g/kg组,Wistar Kyoto大鼠(WKY)为正常对照组,连续灌胃给予相应药物或蒸馏水8 w。每两周测大鼠尾动脉血压;实验末,记录24 h尿量,进行尿渗透压、Na^(+)、K^(+)离子浓度、HE染色、肾脏指数检测;全自动生化仪测定尿液总蛋白(UTP)、β2-微球蛋白(β2-MG);ELISA试剂盒测定尿液水通道蛋白2(AQP2)和血清精氨酸加压素(AVP)、醛固酮(ALD)含量,免疫印迹法检测肾组织AQP2蛋白表达情况。结果:与正常对照组比较,模型对照组大鼠24 h尿量、Na^(+)、Na^(+)/K^(+)比值显著降低(P<0.01),尿液渗透压、K^(+)、UTP、β2-MG、AQP2含量、血清ALD、AVP含量、肾脏指数及肾组织AQP2蛋白表达显著增加(P<0.01)。与模型对照组比较,真武汤两个剂量组给药后能明显降低UTP、β2-MG的含量(P<0.05或P<0.01),改善SHR肾小球病变,明显降低血清ALD、AVP含量(P<0.05或P<0.01),明显促进尿液排出、明显降低渗透压、明显升高Na^(+)/K^(+)比值(P<0.05或P<0.01),显著降低肾组织AQP2蛋白表达和尿液AQP2含量(P<0.01),以真武汤8.4 g/kg组改善最明显。结论:真武汤通过调节AVP/AQP2通路保护高血压肾损伤。

Objective:To investigate the protective effect of Zhenwu Decoction on renal injury in spontaneously hypertensive rats(SHRs)based on the AVP/AQP2 pathway.Methods:Fifty SHRs were randomly divided into the model group,0.67 mg/kg enalapri group,and 2.1 g/kg and 8.4 g/kg Zhenwu Decoction groups,and the Wistar Kyoto(WKY)rats were classified into the normal control group.During the intragastric administration of the corresponding drugs or distilled water for eight weeks,the blood pressure of rats was measured at the tail artery once every two weeks.At the end of the experiment,the 24 h urine volume was recorded,and the urine osmotic pressure,Na^(+)and K^(+)ion concentrations were measured,followed by HE staining and renal index detection.The urine total protein(UTP)andβ2-microglobulin(β2-MG)were determined using an automatic biochemical analyzer.The urine aquaporin 2(AQP2),arginine vasopressin(AVP),and aldosterone(ALD)were determined by ELISA,and the renal AQP2 protein expression by Western Blotting.Results:Compared with the normal control group,the model group displayed significantly decreased 24 h urine volume,Na^(+),and Na^(+)/K^(+)ratio(P<0.01)but increased urine osmotic pressure,K^(+),UTP,β2-MG,AQP2,serum ALD and AVP,renal index,and renal tissue AQP2 protein expression(P<0.01).Compared with the model group,Zhenwu Decoction at both doses significantly diminished UTP andβ2-MG and serum ALD and AVP levels(P<0.05 or P<0.01),improved the glomerular lesions,promoted urine discharge,lowered the osmotic pressure,elevated the Na^(+)/K^(+)ratio(P<0.05 or P<0.01),and down regulated renal AQP2 protein expression and urine AQP2 level(P<0.01),with better outcomes observed in the 8.40 g/kg Zhenwu Decoction group.Conclusion:Zhenwu Decoction protects the kidney from hypertensive injury by regulating the AVP/AQP2 pathway.