Study Of Wenyang Tongmai Fang On Myocardial Ischemia Reperfusion Cell Poptosis In Rats And The Mechanism Of Akt/GSK-3 Beta Pathway

ObjectiveBy studying the effects of Chinese medicines Wenyang Tongmai Fang to apoptosis of myocardial ischemia reperfusion in rats and Akt/GSK-3 beta pathway,to explore the mechanism of the protection of ischemic cell apoptosis in a rat model of reperfusion injury.MethodsThe 70 male SD rats were divided into 4 groups,include: sham group(sham group),ischemia reperfusion group(IR group),Wenyang Tongmai Fang group(WY group)(Wenyang Tongmai Fang is based on Zhishi Xiebai Guizhi Decoction to change),Zhishi Xiebai Guizhi Decoction group(ZS group).They were fed in laboratory a week,and sham group and IR group rats were fed with distilled water,WY group rats were fed with traditional Chinese medicine Wenyang Tongmai Fang,ZS group rats were fed with Zhishi Xiebai Guizhi Decoction.Within 14 days,those rats was made by intragastric administration.Except for sham group,other three groups were received occlusion of left coronary anterior descending artery(LAD),ischemia 30 min,reperfusion 120 min,establish ischemia reperfusion model,by ECG manifestation to judge whether make a successful ischemia reperfusion model.By Evans blue/TTC double staining method to observe the myocardial infarct size.Study about contents of c Tn I,LDH and CK-MB in serum.The pathological changes of myocardial tissue were observed.Myocardial cell apoptosis was detected by TUNEL kit.By Ca2+induced,detection of myocardial mitochondria m PTP if it is opened.Detect the expression Akt,p-Akt,GSK-3?,p-GSK-3?,Caspase-3,Caspase-9,cytochrome C protein by Western-blot method.ResultsCompared with the sham group,the content of c Tn I,LDH,CK-MB(P<0.01)in serum,apoptosis rate of myocardial cells(P<0.01)and expression of Caspase-3 Caspase-9 and cytochrome C(P<0.05)in the model group significantly increased.In the model group,severe swelling of mitochondria(P<0.01),myocardial fiber arrangement disorder,there are a lot of red blood cells and inflammatory cell infiltration.Compared with the model group,WY group and ZS group could significantly reduce myocardial infarct size(P<0.01),decrease the content of c Tn I,LDH,CK-MB(P<0.01),improved myocardial alignment disorder,reduce apoptosis rate of myocardial cells(P<0.01),and effectively inhibit the swelling of myocardial mitochondria(P<0.01),increased expression of p-Akt and p-GSK-3 beta significantly(P<0.01),decreased Caspase-3 Caspase-9 and cytochrome C expression(P<0.05).About the myocardial infarction area,there is no significant difference between them(P>0.05).Compared with ZS group and WY group,content of LDH,c Tn I decreased significantly(P<0.05)and CK-MB no statistical significance(P>0.05)in WY group.And in WY group,myocardial pathological morphology were improved,myocardial cell apoptosis rate and mitochondrial m PTP are inhibited effectively(P<0.05),increase the expression of p-Akt,p-GSK-3 beta(P<0.05),reduce the expression of Caspase-9,Caspase-3,and cytochrome C(P<0.01).ConclusionsCompared with the model group,Wenyang Tongmai Fang and Zhishi Xiebai Guizhi Decoction can inhibit the cell apoptosis of myocardial ischemia reperfusion injury in rats,Compare with Zhishi Xiebai Guizhi Decoction,Wenyang Tongmai Fang can significantly reduce the content of c Tn I and LDH inserum of rats with ischemia reperfusion injury,improved myocardial alignment disorder,reduce apoptosis rate of myocardial cells,inhibition of mitochondrial m PTP opening,increased expression of p-Akt and p-GSK-3 beta,decreased Caspase-3 Caspase-9 and cytochrome C expression.The mechanism of Wenyang Tongmai Fang protection ischemia reperfusion rats may be through the Akt/GSK-3? pathway,inhibition of mitochondrial membrane opening,thereby reducing the release of cytochrome C and activation of Caspase-3.