Study On The Changes Of Saponins And The Protective Effect Of Myocardial Injury Before And After Steaming American Ginseng

Objective:To explore the best high-temperature steaming process of American ginseng,establish HPLC fingerprint of steaming products of American ginseng,and characterize the overall changes of saponins before and after steaming.Further,HPLC and UPLC-MS methods were used to explore the changes of the saponins’chemical components before and after processing American ginseng,with the changes of components as indicators.To explore the protective effect of American ginseng on myocardial injury in rats before and after steaming,and to explore the possible mechanism.Methods:1.Taking the total amount of ginsenosides Rb₁+Re+Rg₁ and the comprehensive scores of appearance traits as evaluation indexes,single factor investigation was conducted and three factors and three levels were screened.BBD response surface experiment was used to study the optimal steaming process of American ginseng2.HPLC fingerprints were established for 16 batches of American ginseng raw products and their corresponding steamed products,and the established fingerprints were compared to analyze the changes of saponins composition fingerprints before and after steaming.3.The content of 16 batches of Radix ginseng and its corresponding steamed products was determined by HPLC.The contents of 10 saponins were identified and calculated by standard products,and the content changes of chemical components before and after steaming of Radix ginseng were analyzed.4.The saponins of American ginseng before and after steaming were analyzed by UPLC-MS method,and Xcalibur 4.0 software was used for analysis.The components were identified by comparing with the reference products and referring to the component mass spectrum information in the literature,and the different components were screened.5.After 30 days of intragastric administration of raw and steamed extracts of American ginseng,ISO was injected subcutaneously to replicate myocardial injury model,serum was taken for Elisa kit determination,and myocardial tissue was taken for HE staining,Masson staining and immunohistochemical Western-Blot assay.Results:1.the optimal process was obtained as follows:the steaming time was 100min,the steaming temperature was 111℃,and the drying temperature was 70℃.The predicted score value was 107.741 based on the above experimental conditions.2.The fingerprints of 16 batches of raw American ginseng and 16 batches of steamed American ginseng were established,in which 21 common peaks were identified and 7components were identified;A total of 22 common peaks were identified by the fingerprint of 16 batches of steamed American ginseng,and 10 of them were identified,showing high similarity between sample groups.3.A HPLC method for simultaneous determination of 10 saponins was established.The results showed that no Rb₂,Rb₃ and 20（S）-Rg₃ were detected in raw American ginseng.After steaming,the contents of Rb₁,Rb₂,Rb₃ and 20（S）-Rg₃ increased,the content of Rd decreased,and the total amount of ten saponins increased.4.A total of 37 saponins were identified from raw and processed American ginseng by UPLC-MS method,21 from raw American ginseng and 30 from steamed American ginseng.There are 14 kinds of ingredients,7 kinds of unique ingredients in raw products and 16 kinds of unique ingredients in steamed products.5.The results of heart weight index,ELisa kit and HE and Masson staining all showed the same results,namely obvious myocardial injury after modeling,which proved the success of modeling.The damage was reduced in the drug administration group,but no significant difference was found in the low-dose group,the effect of the high-dose group was significantly better than that of the low-dose group,and the effect of steamed American ginseng was slightly better than that of raw ginseng.The results of immunohistochemical and WB experiments showed that compared with the blank group,the expression of Bax and Caspase-3 protein increased significantly in the ISO modeling group,and the expression of Bcl-2 decreased significantly.The improvement effect was not significant in the low dose group,and the expression of Bax and Caspase-3 was decreased in the high dose group,while the expression of Bcl-2 was increased.Conclusion:A stable and fast cooking process of American ginseng was established through process optimization,and the changes of saponins before and after cooking were analyzed through the fingerprint of raw and steamed products of American ginseng and the qualitative and quantitative determination of saponins components.It was proved that the species and content of saponins changed significantly before and after processing,which may be the basis for correcting and improving the efficiency of American ginseng after processing.Pharmacological experiments showed that both raw and steamed American ginseng could improve myocardial damage caused by ISO by inhibiting apoptosis,and the effect of steamed American ginseng was better than that of raw American ginseng.