Roudoukou-8 San Of Mongolian Drug Reduces The Myocardial Ischemia Reperfusion Injury Of Rat Through JAK2/STAT3 Pathway

Objective:To study the protective effects of Roudoukou-8 San on myocardial ischemia reperfusion injury（MIRI）of rats and its relationship with tyrosine protein kinase2（JAK2）/signal transducer and activator 3（STAT3）signaling pathway.Methods:（1）60 wistar male rats were randomly divided into 6 groups:sham group,MIRI model group,AG490blocker group,Roudoukou-8 San low dose group,Roudoukou-8 San medium dose group and Roudoukou-8 San high dose group,10 rats in each group.Lesion of left anterior descending coronary was ligated for 30min and reperfused for 24h.Detection of rat ECG through BL-420S biological function experiment system.The contents of lactate dehydrogenase（LDH）,creatine phosphate kinase（CK）and aspartate aminotransferase（AST）in rat serum were tested by fully automatic biochemical analyzer.The contents of catalase（CAT）,superoxide dismutase（SOD）.And glutathione peroxidase（GSH-Px）in rat myocardial tissue were tested by kit method.And the m RNA expressions of JAK2,STAT3,bcl-2 and Bax in myocardial tissue of each group were tested by reverse transcription real-time polymerase chain reaction（RT-q PCR）.the protein expressions of JAK2,p-JAK2,STAT3,p-STAT3,bcl-2and Bax in myocardial tissue of each group were tested by western blotting.（2）Isolation and culture of rat neonatal cardiomyocytes,Hypoxia/reoxygenation injury model of rat neonatal cardiomyocytes was built by sodium dithionite（Na₂S₂O₄）.The morphological changes of cardiomyocytes were observed by an inverted microscope.The vitality of the cells was tested by CCK-8 method.Cell apoptosis degree was tested by hoechst staining and flow cytometry.The contents of LDH,CK and AST in cell culture medium were tested by fully automatic biochemical analyzer.CAT,SOD and GSH-Px in cells were tested by kit method.The m RNA expressions of JAK2,STAT3,bcl-2 and Bax in myocardial cells of each group were tested by RT-q PCR.the protein expressions of JAK2,p-JAK2,STAT3,p-STAT3,bcl-2 and Bax in myocardial cells of each group were tested by western blotting.Result:（1）animal experiment results:Compared with the model group,the contents of CK,LDH and AST in rat serum（P<0.05）could be obviously decreased,and then contents of CAT,SOD and GSH-Px in myocardial tissue（P<0.05）could be obviously increased（P<0.05）.The m RNA expression of JAK2,STAT3 and bcl-2 could be obviously increased（p<0.05）and the m RNA expression of Bax could be obviously decreased（P<0.05）.The protein expression of p-JAK2/JAK2,p-STAT3/STAT3 and bcl-2/Bax could be obviously increased（P<0.05）,while the effects of Roudoukou-8 San were reduced in AG490 blocker group.（2）Cell experiment results:2mmol·L^-1Na₂S₂O₄obviously caused damage of about 50%cells.Compared with the model group,the extracts of Roudoukou-8 San with different concentrations could increase the viability of cells.Besides,the contents of CK,LDH and AST in cell culture medium could be obviously decreased（P<0.05）,and the contents of CAT,SOD and GSH-Px in cells could be obviously increased（P<0.05）.The m RNA expression of JAK2,STAT3 and bcl-2 could be obviously increased（P<0.05）and the m RNA expression of Bax could be obviously decreased（p<0.05）.the protein expression of p-JAK2/JAK2 and p-STAT3/STAT3 could be obviously increased（P<0.05）and the protein expression of bcl-2/Bax could be obviously decreased（P<0.05）,while the effects of Roudoukou-8 San extract were reduced in AG490blocker group.Conclusion:Roudoukou-8 San has definite effect that fight against MIRI.Roudoukou-8 San against MIRI of rats through JAK2/STAT3 signaling pathway.