Experimental Study Of Danshen Tongluo Decoction On Autophagy In Rats With Myocardial Ischemia Reperfusion Injury

Objective: To investigate the effect and mechanism of Danshen Tongluo Jiedu Decoction on autophagy in rats with myocardial ischemia-reperfusion injury(MIRI).Methods:The MIRI model was established by ligation of the left anterior descending coronary artery.The patients were randomly divided into a blank group(K group),a sham operation group(S group),an ischemia-reperfusion group(IR group),and a Danshen Tongluo Jiedu decoction group(DTJD group),autophagy inhibitor group(3-MA group),10 per group.Except for the blank group and the sham operation group,the rats in the other groups were subjected to ligation of the left anterior descending coronary artery for ischemia for 30 min and reperfusion for 30 min to prepare a myocardial ischemia-reperfusion model.Observation of myocardial ultrastructure by electron microscopy.Determination of myocardial injury markers using an automated biochemical analyzer: plasma creatine kinase isoenzyme MB(CK-MB)and serum lactate dehydrogenase(LDH).Detection of cellular oxidative stress levels by ELISA: malondialdehyde(MDA)levels and superoxide dismutase(SOD)activity.The expression of LC3 protein in myocardial tissue of each group was determined by immunohistochemistry.Detection of Myosin Beclin-1 mRNA expression by real-time quantitative reverse transcription-polymerase chain reaction(RT-PCR).Western blot was used to detect the autophagy gene levels in each group:Atg5,Beclin-1 and LC3??/LC3? protein expression.Results: Electron microscopy showed that DTJD group could regulate autophagy of myocardial cells in rats with ischemia-reperfusion injury,protect myocardial cell structure and reduce myocardial ischemia-reperfusion injury.There was no significant difference between group C and group S(P>0.05).Compared with group S,the levels of LDH and CK-MB in serum of IR group were significantly increased(P<0.01);the level of SOD in myocardial tissue was significantly decreased.MDA levels were significantly increased(P<0.01);Beclin-1 mRNA expression was increased in myocardial tissue(P<0.01);myocardial autophagy proteins Atg5,Beclin-1 and LC3??/LC3? were significantly increased(P<0.01);Compared with the DTJD group and the 3-MA group,the levels of CK-MB and LDH in the DTJD group and the 3-MA group were significantly decreased(P<0.01),the SOD level in the myocardial tissue increased,and the MDA level decreased(P<0.01).The expression of Beclin-1 mRNA in the myocardial tissue decreased.<0.01);the expression of autophagy proteins Atg5,Beclin-1 and LC3??/LC3? was decreased(P<0.01).Conclusion:1.Danshen Tongluo Jiedu Decoction can significantly reduce the damage of myocardial cell mitochondria,improve the pathological state of myocardial tissue,and affect the morphology of autophagosomes.2.Danshen Tongluo Jiedu Decoction can significantly up-regulate SOD and decrease MDA,reduce mitochondrial damage,and weaken oxidative stress,thereby down-regulating the ratio of Atg5 protein to Beclin-1 protein and LC3??/LC3?,and inhibiting autophagy in MIRI.Play a role in protecting damaged cardiomyocytes.3.Danshen Tongluo Jiedu Decoction can reduce the autophagy response of MIRI by lowering the ratio of Atg5 protein and Beclin-1 protein and LC3??/LC3?,and reduce the levels of CK-MB and LDH,thus reducing the damage of rat cardiomyocytes.