Intervention And Protective Effects Of Ginsenoside Rb1 Combined With PRDX6 On Myocardial Injury

Objective:In this study,we investigated the protective effects of ginsenoside Rb1（Gs-Rb1）and PRDX6 on myocardial injury models at in vitro and in vivo levels using isoproterenol（ISO）-induced myocardial injury cells,rat models,and tried to elucidate their metabolic mechanisms to provide new ideas for the treatment of myocardial injury in clinical settings.Methods:At the in vitro level,H9c2 rat cardiomyocytes were resuscitated,passaged and cultured into control,model,Gs-Rb1,PRDX6 and combination groups:equal volume of PBS was added to the culture medium of control group;equal volume of PBS was added to the culture medium of model group;0.4 mg/m L Gs-Rb1 was added to the culture medium of Gs-Rb1 group;PRDX6medium was added to 1.0 mg/m L PRDX6;0.4 mg/m L Gs-Rb1 and 0.8 mg/m L PRDX6 were added to the combined group medium;the groups were pretreated for 2 h and then added to ISO-injured cardiomyocytes for 24 h.Cell activity was measured by tetrazolium salt（MTT）colorimetric assay;cell ROS content was determined by the kit method;by flow cytometry,apoptosis was detected;as well as the expression levels of intracellular superoxide dismutase（SOD）,glutathione peroxidase（GPX）,and malondialdehyde（MDA）and LDH were measured.At the in vivo level,male SD rats were injected intraperitoneally with 80 mg/kg ISO for 7consecutive days to establish a myocardial injury model（control group was injected with an equal volume of saline）,and after verification of successful model establishment,the rats were randomly divided into control group,model group,Gs-Rb1 group,PRDX6 group,combination group and positive control group（n=6）for 14 days of continuous treatment:control rats were injected intraperitoneally with an equal volume of rats in the control group were injected intraperitoneally with an equal volume of saline;rats in the model group were injected intraperitoneally with an equal volume of saline;rats in the Gs-Rb1 group were injected intraperitoneally with 20 mg/kg Gs-Rb1;rats in the PRDX6 group were injected intraperitoneally with 2.0μg/kg PRDX6;rats in the combination group were injected intraperitoneally with 20 mg/kg Gs-Rb1 and 2.0 mg/kg PRDX6;rats in the positive control group were injected intraperitoneally with coenzyme Q₁₀at the end of the treatment period,rats in each group were examined by electrocardiogram,blood was collected under anesthesia and heart tissues were stained with hematoxylin-eosin（H&E）to visualize the changes in histomorphology of the myocardium and Masson staining to observe the morphological changes of myocardial fibers in rats.The expression levels of superoxide dismutase（SOD）,catalase（CAT）,glutathione peroxidase（GPX）,creatine kinase（CK）,malondialdehyde（MDA）,and lactate dehydrogenase（LDH）were measured by the kit.Transcriptomic sequencing analysis was performed on the model and combination treatment groups,and Western blot method was further validated.Results:In vitro level:MTT results showed that cell proliferation viability was increased in the Gs-Rb1 and PRDX6 groups compared to the model group（P<0.05）;cell proliferation viability was significantly increased in the combination group（P<0.01）.The results of apoptosis showed that the apoptosis rate was reduced in the Gs-Rb1 and PRDX6 groups compared with the model group（P<0.05）,and significantly reduced in the combined group（P<0.01）.The results of cell-related biochemical indexes showed that GPX and SOD activities were significantly increased and MDA content was decreased by combined treatment（P<0.05）.At the in vivo level:the results of rat electrocardiogram showed that the ST segment was significantly elevated and the heart rate was deranged in the model group compared with the control group;the situation was improved in all groups compared with the model group.The results of H&E staining showed that the situation was improved in the Gs-Rb1 group and PRDX6 group compared with the situation of myocardium with broken and disordered fibers in the model group,and the situation was significantly improved in the combined group.The Masson staining showed that a large amount of collagen deposition and myocardial fibrotic changes were seen between the myocardial tissues of rats in the model group,and the collagen fibers were significantly reduced in the combined group.The results of biochemical indexes showed that the serum CAT,GPX,SOD,LDH,MDA and CK were gradually normalized after treatment;Transcriptomic findings suggest that Gs-Rb1 and PRDX6 may ameliorate myocardial injury by regulating signaling pathways such as SIRT3 and PPAR;Western Blot showed that the Bcl-2/Bax ratio was increased（P<0.05）,the expression level of SIRT3 protein was significantly increased（P<0.01）and the expression level of NF-κB protein was decreased in the myocardial tissues of the treated rats（P<0.05）.Conclusion:1.Gs-Rb1 and PRDX6 can effectively improve cardiac function,reduce collagen deposition between myocardial fibers and restore myocardial elasticity in rats with myocardial injury.2.The protective effects of Gs-Rb1 and PRDX6 on myocardial tissue in rats with myocardial injury may be achieved by regulating myocardial tissue oxidative stress,reducing apoptosis and decreasing the expression of inflammatory response,and their main signaling pathways are SIRT3 and PPAR,etc.3.Gs-Rb1 showed synergistic effects with PRDX6.