Comparative Study On The Chemical Constituents And The Prevention And Treatment Of Acute Renal Injury Induced By Cisplatin Of Ginger-processed American Ginseng And American Ginseng

American ginsenoside（Panax quinquefolium L.,AG）is a perennial herb belonging to the genus Ginseng of the acanthopanax family.It has the effects of supplementing qi,nourishing yin,clearing heat,and promoting fluid production.Due to its cold and cool nature,its clinical application is limited.Many ancient books record different processing methods of American ginseng to improve its cold and cool characteristics."Wu Jutong Medical Record"records a large number of processing methods of American ginseng,including American ginseng fried with ginger juice,ginger juice yellow and ginger fried;Ginger-processed American ginseng（GPAG）has been included in Volume 1 of the"Processing Specifications for Traditional Chinese Medicine Pieces in Jilin Province"（2020 Edition）.Although clinical practice and pharmacological studies have shown that processed American ginseng with ginger can enhance its effects of supplementing qi,moistening dryness,and strengthening qi,the changes in chemical components of American ginseng caused by processing and the impact on the changes in blood components of American ginseng are still unclear.Therefore,in this study,UPLC-Q-TOF-MS^E technology was used to systematically study the changes in chemical components of American ginseng after processing with ginger and the impact on the blood inflow components of American ginseng.Based on the blood components of GPAG,the potential intervention disease types of GPAG were predicted using network pharmacology methods.Based on this,the mechanism of prevention and treatment of cisplatin induced acute renal injury by GPAG quinquefolium was deeply studied using pharmacodynamic and untargeted metabolomics methods.The main research contents include the following aspects:1.Comparative Analysis of Chemical Constituents of GPAG and AGIn order to clarify the impact of ginger processing on the chemical composition changes of American ginseng,this study systematically analyzed the chemical components of GPAG and AG using UPLC-Q-TOF-MS^E technology,and identified 85compounds including triterpenoid saponins and gingerols.Through comparative analysis of the types of compounds in GPAG and AG,it was found that GPAG had four more components derived from ginger than AG,including 6-Gingerole,6-Gingerol,8-Gingerol,and 10-Gingerol,suggesting that the ginger processing process had no impact on the types of chemical components in American ginseng in vitro.Using PCA and OPLS-DA multivariate statistical analysis methods,the differential components and relative content changes of GPAG and AG were analyzed.The results showed that there were significant differences in the content of 52 compounds（P<0.05）between GPAG and AG,including 24 compounds such as Ginsenoside Rb₃,Ginsenoside Rb₁,and Ginsenoside Rf.The content of Ginsenoside decreased significantly after ginger processing.It is speculated that Ginsenoside may interfere with the metabolic transformation process of chemical components of Ginsenoside during the process of moistening and drying with ginger juice.Correlation analysis was used to analyze the correlation between the different components of GPAG.The results showed that there was a correlation between the components of ginger and AG,further indicating that the components of ginger juice interfered with the metabolic transformation of American ginseng saponins during the ginger processing process.Quantitative analysis was conducted on 12 main compounds out of 52 different compounds,and the content determination results were consistent with the relative content analysis results.This study clarified the impact of ginger processing on the composition changes of American ginseng,laying a material foundation for the participation of GPAG in the differential analysis of the blood components of AG.2.Comparative Analysis of Blood Ingredient between Ginger and American GinsengIn order to explore the effect of ginger processing on the blood inflow components of American ginseng,this study further explored the differences and changes in the blood inflow components of GPAG and AG based on clarifying the in vitro components of GPAG.Based on the UPLC-Q-TOF-MS^E method,the pharmaceutical components in the plasma of rats administered with GPAG and AG ginseng by gavage were analyzed and identified.The results showed that there were 37 blood infused components identified in the GPAG group,and 27 blood infused components in the AG group.Compared with the results of in vitro component analysis,7 chemical components of American ginseng were only identified in plasma.By comparing the components of Ginsenoside Rh₇,Ginsenoside Rh₈,Ginsenoside Rh₁₀,Ginsenoside C-K and other rare ginsenosides,as well as 6-Gingerdione,6-Gingerol,8-Gingerol,and 10-Gingerol,which are only identified in the GPAG group,it is speculated that the components of GPAG can promote the absorption and metabolism of macromolecular ginsenosides in the body after absorption into the blood,thereby converting them into rare saponins.Further analysis of the potential metabolic transformation rules among blood entering components showed that American ginseng processed with ginger can accelerate the hydrolysis and metabolism of various ginsenosides,resulting in the final metabolism of PPD and PPT type ginsenosides into aglycones.Through the analysis of components in vitro and in vivo,it can be seen that ginger processing can promote the absorption of ginsenosides into the blood and play a better role in vivo.This study lays the foundation for the next comparative study of network pharmacology based on the blood components of GPAG and AG.3.Comparative Study on the Mechanism of GPAG and AG in Preventing and Treating Cisplatin-induced Acute Renal Injury Based on Network PharmacologyIt has been reported in the literature that American ginseng has a good preventive and therapeutic effect on cisplatin induced acute renal injury.In order to deeply explore the potential pharmacological effects and mechanisms of GPAG,this study uses a targeted network pharmacological method based on blood inflow components to systematically predict the potential disease intervention and pharmacological mechanisms of GPAG and AG.Disease enrichment prediction analysis based on Swiss Target Prediction and Dis Ge Net databases found that GPAG have an intervention effect on kidney damage.Combined with clinical application and literature reports,AG has a good preventive effect on cisplatin induced acute renal injury.Further exploration using network pharmacology methods found that GPAG have more potential targets for preventing and treating cisplatin induced acute renal injury than AG.The potential targets in the GPAG group are mainly enriched in key signal pathways related to cisplatin-induced acute renal injury,including PI3K-Akt signal pathway,JAK-STAT signal pathway,NF-kappa B signal pathway,arachidonic acid metabolism pathway,and necrotic apoptosis signal pathway.Targets such as SRC,PIK3R1,HSP90AA1,PIK3CA,MAPK1,and PTPN11 are not only closely related to the above differential pathways,"It is also a protein with high correlation in PPI analysis,and these target proteins may be key targets for GPAG in preventing and treating cisplatin-induced acute renal damage.".Through molecular docking methods,the interaction between the 10different blood entry compounds and the core target proteins of GPAG and AG was verified.The results showed that stable conformations could be formed between the main differential components and the core target proteins of GPAG and AG.It is speculated that these components may be the key components of GPAG that have better prevention and treatment effects on cisplatin-induced acute renal injury than AG.The research results lay a foundation for the comparative study of the pharmacological effects and mechanisms of GPAG on cisplatin induced acute renal injury.4 Comparative Study on the Mechanism of Effects of GPAG and AG on Cisplatin-induced Acute Renal Injury in Rats Based on MetabonomicsIn order to deeply explore the pharmacological effects and mechanisms of GPAG on cisplatin induced acute renal injury in rats,a comparative study was conducted on the pathophysiology and endogenous metabolic profile changes of the established model of cisplatin induced acute renal injury in rats based on pharmacodynamic evaluation and non-targeted metabonomics in urine and serum.The results showed that both GPAG could reduce BUN,Cr,Pro,and TNF in the serum of cisplatin-induced AKI model rats-α、IL-6、IL-1βAnd MDA levels,improve the activities of SOD and GSH in kidney tissue,and improve the pathological changes of kidney tissue caused by AKI.Studies have shown that both GPAG have a certain protective effect on cisplatin induced acute renal injury,and GPAG have a stronger pharmacological effect.Based on clarifying the pharmacological effects,a comparative study was conducted on the metabolic mechanisms of GPAG in preventing and treating cisplatin induced acute renal injury using non-targeted metabonomics techniques in urine and serum.Compared with the control group,29 and 24 potential biomarkers in the urine and serum of cisplatin-induced acute renal injury model rats were screened for significant changes,respectively.In the AG group,11 markers in the urine and 4 markers in the serum were significantly regulated,while in the GPAG group,25 markers in the urine and 14markers in the serum were significantly regulated,involving the citric acid cycle（TCA cycle）,phenylalanine metabolism,tryptophan metabolism There are 9 metabolic pathways involved in bile acid biosynthesis,purine metabolism,taurine and sub taurine metabolism,arginine biosynthesis,lysine biosynthesis,and fatty acid metabolism.It is suggested that GPAG may improve cisplatin induced acute renal injury by regulating these metabolic pathways.This study elucidated the possible mechanism of GPAG in preventing and treating cisplatin-induced acute renal injury from the perspective of non-targeted urine and serum metabolomics,laying a foundation for further research on GPAG in preventing and treating cisplatin-induced acute renal injury.To sum up,In this study,UPLC-Q-TOF-MS^E technology,network pharmacology,molecular docking methods,pharmacodynamics,and metabolomics were used to conduct a comparative study of the chemical components in vitro and in vivo of GPAG and AG,as well as their mechanisms of preventing and treating cisplatin induced acute kidney injury,providing theoretical basis for revealing the scientific connotation of the effects of processed Ginger American Ginseng on supplementing qi,moistening dryness,and strengthening qi,Meanwhile,it laid a research foundation for guiding the clinical application of GPAG.