Protective Effect And Mechanism Of Ginseng Blood Composition On Cardiomyocytes In Shenfu Decoction

Objective:To explore the protective effect of ginseng blood composition on H9C2cardiomyocytes in Shenfu decoction,and to study its specific protective mechanism in depth.Methods:1 Study on the protective effect and mechanism of anti-hypoxia reoxygenation induced myocardial cell injury in ginseng blood components in Shenfu decoctionThe hypoxic reoxygenation injury model of H9C2 Cardiomyocytes was established,which was compatible according to the proportion of ginseng blood components in the Shenfu decoction detected in the early stage of the laboratory.Divided into C Group,H/R Group,SFP Group,SF Group,Rg1 Group,Rg2 Group,Rb1 Group,Re Group total of 8 groups to carry out experiments.The growth status of cells was observed by inverted fluorescence microscope,and the survival rate of cells was detected by MTT method.Enzyme marker detection of LDH leakage rate to determine the degree of cell damage.The active components of ginseng Rg1 and Rg2 were screened.The optimum concentration of ginsenoside Rg1 and Rg2 protection was also screened.Then the study of protective mechanism is divided into 6groups,C Group,H/R Group,SFP Group,SF Group,Rg1 Group,Rg2 Group.The degree of myocardial cell injury was determined by the detection of MDA and SOD of oxidative stress index by enzyme marker.The content of intracellular reactive oxygen ROS was observed by inverted fluorescence microscope,the concentration of intracellular Ca²⁺was detected by fluorescence spectrophotometer,and the apoptosis rate was detected by TUNEL method.2 Study on the protective effect and mechanism of anti-H₂O₂ induced myocardial cell injury in ginseng blood components in Shenfu decoctionThe H₂O₂ injury model of H9C2 Cardiomyocytes was established,which was compatible according to the proportion of ginseng blood components in the Shenfu decoction detected in the early stage of the laboratory.Divided into C Group,H₂O₂ Group,SFP Group,SF Group,Rg1 Group,Rg2 Group,Rb1 Group,Re Group total of 8 groups to carry out experiments.The damage degree of cardiomyocytes was detected by MTT and LDH index,and whether the blood composition of ginseng in Shenfu decoction had protective effect on the myocardial cells of H₂O₂ injury.The active components of ginseng Rg1 and Rg2 were screened.The optimum concentration of ginsenoside Rg1 and Rg2 protection was also screened.Then the study of protective mechanism is divided into 6 groups,C Group,H₂O₂ Group,SFP Group,SF Group,Rg1 Group,Rg2 Group.From two aspects of oxidative stress and apoptosis,the main research was carried out,and the oxidative stress index MDA and SOD were tested to determine its oxidation damage ability and antioxidant damage ability.The content of ROS,Ca²⁺concentration and apoptosis rate of intracellular reactive oxygen species were detected,and the mechanism of apoptosis was studied.3 Study on the protective effect and mechanism of the anti-Isoprenaline（ISO）induced myocardial cell injury in ginseng blood components in Shenfu decoctionThe ISO damage model of H9C2 Cardiomyocytes was established,which was compatible according to the proportion of ginseng blood components in the Shenfu decoction detected in the early stage of the laboratory.Divided into C Group,ISO Group,SFP Group,SF Group,Rg1 Group,Rg2 Group,Rb1 Group,Re Group total of 8 groups to carry out experiments.Three indexes of MTT and LDH were detected to determine the degree of myocardial cell injury,and whether it had protective effect on it after administration.The active components of ginseng Rg1 and Rg2 were screened.The optimum concentration of ginsenoside Rg1 and Rg2 protection was also screened.Then the study of protective mechanism is divided into 6 groups,C Group,ISO Group,SFP Group,SF Group,Rg1 Group,Rg2 Group.The content of ROS,Ca²⁺concentration and apoptosis rate of intracellular reactive oxygen species were detected,and the mechanism of apoptosis was studied.Results:1 Study on the protective effect and mechanism of anti-hypoxia reoxygenation induced myocardial cell injury in ginseng blood components in Shenfu decoctionHypoxic reoxygenation injury can significantly reduce the survival rate of H9C2cardiomyocytes（P<0.001）and increase the incidence（P<0.001）of lactate dehydrogenase LDH.Each dosing group could improve the survival rate of cardiomyocytes and reduce the incidence of LDH leakage,and the effect of ginseng into blood composition Rg1 and Rg2（p<0.001,p<0.01）in Shenfu Decoction was better than that of Rb1 and Re（P<0.05）.It is indicated that each drug group has protective effect on myocardial cells injured by hypoxic reoxygenation H9C2,and the protective effect of ginseng into blood composition Rg1 and Rg2 in Shenfu decoction is stronger than that of Rb1 and Re.The optimum concentration of ginsenoside Rg1 and Rg2 protective effect was screened,and the results showed that ginsenoside Rg1 with concentration of 5μg/ml and 1μg/ml with concentration of ginsenoside Rg2 had the best protective effect on injured cardiomyocytes（P<0.01）.Therefore,the subsequent experimental selection of Ginsenoside Rg1 with a concentration of 5μg/ml and1μg/ml of ginsenoside Rg2 the protective mechanism of H9C2 myocardial cell injury was studied in depth.Hypoxic reoxygenation can significantly improve the MDA content（P<0.001）in H9C2 cardiomyocytes,and the MDA content in cells can be significantly reduced in each drug delivery Group（P<0.001）.Hypoxic reoxygenation can significantly reduce the SOD content of superoxide dismutase（P<0.001）,each drug group can improve the intracellular SOD content（P<0.01,P<0.05）to varying degrees.Hypoxic reoxygenation can significantly improve the Ca²⁺concentration（P<0.01）in cytoplasm,and each drug group has a protective effect on cell membranes,which can reduce the concentration of Ca²⁺in cytoplasm（P<0.01,P<0.05）to varying degrees.Hypoxic reoxygenation can significantly increase the apoptosis rate of cardiomyocytes（P<0.01）.Hypoxia/reoxygenation can increase the content of ROS in cells,and each drug group can reduce the content of ROS in cells.The drug delivery Group has protective effect on cardiomyocytes,which can reduce the apoptosis rate of cardiomyocytes（P<0.01,P<0.05）.2 Study on the protective effect and mechanism of H₂O₂ induced myocardial cell injury by ginseng blood components in Shenfu decoctionH₂O₂ can significantly reduce the survival rate of cardiomyocytes（p<0.001）,Shenfu decoction Serum,Shenfu decoction,ginseng into the blood component Rg1,ginseng into the blood component Rg2 can significantly improve the survival rate of cardiomyocytes（p<0.001）.By detecting LDH to judge the degree of myocardial cell injury,H₂O₂ can significantly reduce the LDH leakage rate（p<0.001）of cardiomyocytes,and the LDH leakage rate（p<0.001）of cardiomyocytes can be significantly improved by using Shenfu decoction serum,Shenfu decoction,ginseng into the blood component Rg1 and ginseng into the blood component Rg2.The effect of ginseng blood composition on H₂O₂ induced myocardial cell injury in Shenfu Decoction was explained.The degree of myocardial cell injury was further judged by the detection of MDA and SOD.The MDA content（p<0.001）of cardiomyocytes could be significantly increased,and the MDA content（p<0.001）of cardiomyocytes was significantly reduced in the Shenfu decoction serum of H₂O₂,Shenfu decoction,ginseng into the blood component Rg1,ginseng into the blood component Rg2.H₂O₂ can significantly reduce the SOD content of cardiomyocytes（p<0.001）,and the SOD content（p<0.01,p<0.05）of cardiomyocytes can be improved by including Shenfu decoction serum,Shenfu decoction,ginseng blood composition Rg1 and ginseng blood composition Rg2.H₂O₂ can significantly increase the content of[Ca²⁺]i content（P<0.01）and reactive oxygen ROS in the cytoplasm of cardiomyocytes.The content of[Ca²⁺]i content（p<0.05）and reactive oxygen ROS in the cytoplasm of cardiomyocytes can be increased by the Shenfu decoction serum,Shenfu decoction,ginseng blood composition Rg1 and ginseng into the blood composition Rg2.H₂O₂can significantly increase the apoptosis rate of cardiomyocytes（P<0.001）.The apoptosis rate（p<0.01,p<0.05）of myocardial cells can be improved by the Shenfu decoction serum,Shenfu decoction,ginseng blood composition Rg1 and ginseng blood composition Rg2.3 Study on the protective effect and mechanism of anti-Isoprenaline（ISO）induced myocardial cell injury in ginseng blood components in Shenfu decoctionISO can significantly reduce the survival rate of cardiomyocytes（p<0.001）,Shenfu decoction Serum,Shenfu decoction,ginseng into the blood components Rg1,ginseng into the blood component Rg2 can significantly improve the survival rate of cardiomyocytes（p<0.001）,the myocardial cells have a protective effect.ISO can significantly reduce the LDH leakage rate（p<0.001）of cardiomyocytes,Shenfu decoction Serum,Shenfu decoction,ginseng into the blood composition Rg1,ginseng into the blood component Rg2 can significantly improve the myocardial cell LDH leakage rate（p<0.001）.MDA and SOD are typical oxidative stress indexes,ISO can significantly improve the MDA content of Cardiomyocytes（p<0.001）,Shenfu decoction Serum,Shenfu decoction,ginseng into the blood components Rg1,ginseng into the blood component Rg2 significantly reduce the MDA content of Cardiomyocytes（p<0.001）.ISO can significantly reduce the SOD content of cardiomyocytes（p<0.001）,Shenfu decoction Serum,Shenfu decoction,ginseng into the blood components Rg1,ginseng into the blood components Rg2 can improve the SOD content of cardiomyocytes（p<0.01,p<0.05）.ISO can significantly improve the[Ca²⁺]i content（P<0.01）in cytoplasmic cytoplasm of cardiomyocytes.The[Ca²⁺]i content（p<0.01,p<0.05）in the cytoplasm of cardiomyocytes can be increased by the Shenfu decoction serum,Shenfu decoction,ginseng blood composition Rg1 and ginseng into the blood composition Rg2.ISO can improve the content of ROS in myocardial fine reactive oxygen species.The content of reactive oxygen ROS in cardiomyocytes can be reduced by the Shenfu decoction serum,Shenfu decoction,ginseng blood composition Rg1 and ginseng blood composition Rg2.ISO can significantly increase the apoptosis rate of cardiomyocytes（P<0.001）.The apoptosis rate（p<0.01,p<0.05）of myocardial cells can be improved by the Shenfu decoction serum,Shenfu decoction,ginseng blood composition Rg1 and ginseng blood composition Rg2.Conclusion:Ginseng Blood composition in Shenfu decoction has protective effect on injured cardiomyocytes,and its protection may be related to protecting cell integrity,inhibiting oxidative stress,inhibiting calcium overload and inhibiting apoptosis.