Effects Of Wenyangtongmaifang Via AC/cAMP Pathway On Ischemia Reperfusion Myocardium Of Rats AQP

ObjectiveTo deserve the effect of Wenyangtongmai Decoction on myocardial ischemia reperfusion injury(myocardial ischemia reperfusion injury,MIRI)rats water channel protein AQP,and to explore its possible mechanism.Methods36 SD rats were randomly divided into 4 groups,sham operation group,model group,Wenyangtongmai Decoction group(Wen Yangtongmai is based on Zhishi Xiebai Guizhi and ren shen、sanqi),Zhishi Xiebai Guizhi Decoction group,9 rats in each group.The distilled water is fed for one week and then grouped for 14 days.Feeding methods: The sham operation group only opened the chest and did not ligation at the left coronary artery.The other three groups were thoracotomy and ligation of the left anterior descending coronary artery(LAD)of the left coronary artery(30min),and then the 120 min reperfusion model was established by reperfusion.Specific methods as following: The sham operation group only opened the chest and did not ligation at the left coronary artery.The other three groups were thoracotomy and ligation of the left anterior descending coronary artery(LAD)of the left coronary artery(30min),and then the 120 min reperfusion model was established by reperfusion.Detection the changes of LDH and c Tnl in serum,Observation of pathological changes of myocardium in rats with HE,the contents of AC and c AMP in myocardial tissue were made by Elisa,immunohistochemical was used to check AQP1,AQP4 expression,AQP1 and AQP4 m RNA expression by Real-Time PCR,PKA,P-PKA,CREB,P-CREB,AQP1,AQP4 changes of myocardial protein by Western-blotting.ResultsCompared with sham operation group,the model group’s myocardial fibers were wavy,cell vacuolizated and a large number of red cells infiltrated,the contents of LDH,c Tnl in serum were significantly increased(P<0.01),the contents of AC,c AMP in plasma were decreased significantly(P<0.01),the expression of P-PKA,P-CREB,AQP1,AQP4 protein were decreased(P<0.01),the expression of AQP1,AQP4 m RNA were increased(P<0.01).Compared with the model group,the two administration groups can significantly improve the myocardial fibers and reduce the accumulation of inflammatory cells and change the morphology of the cells,reduced the contents of LDH,c Tnl in serum and the contents of AC,c AMP in plasma(P<0.01),the protein of P-PKA,AQP1,AQP4 and AQP1,AQP4 m RNA expression were significantly decreased(P<0.01).Compared with Zhishi Xiebai Guizhi Decoction group,Wenyangtongmai Decoction group can improve myocardial pathological tissue more efficiently,reduce cell swelling,decreased the contents of LDH,c Tnl in serum and the contents of AC,c AMP in plasma(P<0.01),reduced the expression of P-PKA,AQP1,AQP4 protein and AQP1,AQP4 m RNA(P<0.01).The protein expression of P-CREB in the two drug groups did not change.ConclusionsWenyangtongmai and Zhishi Xiebai Guizhi Decoction can reduce myocardial pathological changes and improve the content of LDH,c Tnl in serum,improve the myocardial ischemia reperfusion injury effectively,Wenyangtongmai is more better.By decreasing the contents of AC and c AMP in plasma and the protein expression of P-PKA,AQP1,AQP4 and AQP1,AQP4 m RNA,the swelling of cells was alleviated.So indicated that Wenyangtongmai protection decreased ischemia-reperfusion injury may related with the down-regulation of AC/c AMP signaling pathway,and inhibited AQP1,AQP4 m RNA and protein expression.