| Objective:American ginseng（Panax quinquefolius L.）is a perennial herbaceous plant belonging to the family Araliaceae and is a precious traditional Chinese medicine widely used in clinical treatment of cardiovascular diseases.To enhance the comprehensive utilization of American ginseng and expand its medicinal channels,research has been conducted on the anti-myocardial damage activity of American ginseng flower,which are the non-medicinal parts of American ginseng.The study selected the chemical components of AGF and has theoretical value and guiding significance for the development and utilization of non-medicinal parts of American ginseng resources.Methods:In this study,a comparative analysis of the pharmacological substance basis of American ginseng flower was conducted based on the requirements of the 2020 edition of the Chinese Pharmacopoeia for the appearance,thin-layer identification,and content determination of American ginseng.Ten different chemical components of American ginseng flower were extracted using high-temperature and high-pressure treatment to increase the transformation of rare ginsenosides and extraction with polar solvents including n-hexane,chloroform,ethyl acetate,n-butanol,and water.The MTT and DCFH-DA were used to determine the cell viability and reactive oxygen species（ROS）level of H2O2-induced H9c2 myocardial cells damage model,respectively,to screen for active components of American ginseng flower that have anti-myocardial damage activity in vitro.The study established a myocardial injury mouse model by intraperitoneally injecting doxorubicin（DOX）into SPF-grade Kunming mice.The model mice were randomly divided into eight groups:normal group,model group,verapamil group（positive drug）,PEE group,before and after high-temperature and high-pressure treated American ginseng flower,and American ginseng root group.After 14 days of oral administration,the mice were examined for appearance,weight,and other indicators.The heart tissue of the mice was stained with H&E to observe the morphology of myocardial tissue cells.Blood was collected for enzyme-linked immunosorbent assay（ELISA）to determine the levels of LDH,CK,CK-MB and AST.The content of ginsenosides,proteins,and flavonoids in PEE was determined using the glacial acetic acid-perchloric acid,total protein quantification test kit,and aluminum nitrate-sodium sulfite,respectively.The pharmacological substance basis of the anti-myocardial damage activity of PEE was explored using liquid chromatography-mass spectrometry（LC-MS）technology.Results:Through an investigation of the pharmacological components of American ginseng flower,it was found that they are rich in ginsenosides,providing a theoretical and material basis for future research.Among the components extracted by gradient extraction,the American ginseng flower ethyl acetate extract（PEE）obtained through high temperature and high pressure treatment showed good ability to increase the cell viability of H9c2myocardial cells induced by H2O2 damage and reduce cellular ROS levels.In mouse model of DOX-induced myocardial injury,treatment with PEE effectively reduced the HMI value and the levels of myocardial enzymes and AST in the serum,protecting myocardial cell damage.H&E staining results showed that PEE treatment effectively alleviated myocardial cell apoptosis,maintained the original structural morphology of myocardial cells,and promoted the growth of myocardial cell numbers.Chemical analysis showed that compared to the pre-high temperature and high pressure American ginseng flower ethyl acetate extract（OEE）of the same polarity,PEE had a similar chemical composition,with nearly identical total ginsenoside content,slightly lower total protein and flavonoid content.The results of LC-MS analysis showed that compared to OEE,PEE had a significant increase in the content of ginsenosides Rk1 and Rg5 and a certain proportion of these components,suggesting that the anti-myocardial injury activity of PEE may be due to the presence of ginsenosides Rk1 and Rg5.Conclusion:The American ginseng flower contain abundant ginsenoside components similar to those found in American ginseng,providing a theoretical and material basis for their potential medicinal value.The PEE may serve as a novel active component for treating myocardial damage,as it effectively alleviates and treats H2O2-induced H9c2myocardial cell damage and DOX-induced mouse myocardial injury,with the ginsenosides Rk1 and Rg5 possibly serving as the active medicinal components responsible for its anti-myocardial damage effects.These findings suggest that the ginsenoside resources found in non-medicinal parts of American ginseng are worth further exploration and utilization,providing a new direction and research method for mitigating the shortage of American ginseng resources and improving their overall utilization. |
