Study Protopanaxadiol-type Ginsenosides Metabolism By Gut Microbiota

Objective:Since December of 2019,when the outbreak of pneumonia caused by COVID-19 in Wuhan,Hubei and other areas,traditional Chinese medicine has played an important role in this treatment of epidemic situation.Referenced to Ginseng Antiphlogistic Powder from Taiping Huimin Heji Ju Fang,ginseng comes with a function to balance body inside.By oral administration of ginseng or ginseng product,gut microflora and ginsenoside components will interact each other to complete the metabolic absorption process and then produce effects on human body in intestine,which is the main absorption site of human body.While being main component of ginsengoside,protopanaxadiol?PPD?-type ginsenosides offer multi functions.The purpose of this paper is to investigate the metabolic effect of gut microflora on PPD-type ginsenosides and the effect of PPD-type ginsenosides on gut microflora.Method:1.Production of glycol ginsenosides:Extract total ginsenosides from dried ginseng by decocting,then separate and purify PPD-type ginsenosides by macroreticular resin adsorption.2.Transformation and pathway analysis of PPD-type ginsenosides in gut microflora:Establish an analytical method for the identification of saponin in transformed samples by RRLC-Q-TOF MS/MS,consider fragmentation pattern of main components,compare with Reference Standard,and reference to the relevant literature to qualitatively identify the saponin components in samples.3.Research of PPD-type ginsenosides conversion and transformed efficiency:Establish quantitative determination of saponin content in convert sample,process experimental data in batch,then calculate content of every saponins in sample.4.Effect of PPD-type ginsenosides on gut microflora:Based on 16S rDNA gene sequencing technique,analysing effect of PPD-type ginsenosides on gut microflora by indexes of microflora diversity,gut microflora and so on.Results:1.Extracting and purifying of PPD-type ginsenosides:Macroreticular resin:D101;Sample concentration:15 mg/mL;Absorption time:12 h;Mobile phase:75%ethyl alcohol.2.Identification of PPD-type ginsenosides metabolite and fitting of metabolic pathway:Ginsenoside Rb₁ converts to 7 metabolites,with the pathway of Rb₁?Rd?F₂?CK?PPD,Rb₁?Gyp-XVII?F₂?CK?PPD,Rb₁?Rd?Rg₃?Rh₂?PPD;Ginsenoside Rc converts to 7 metabolites,with the pathway of Rc?Rd?F₂?CK?PPD,Rc?Rd?Rg₃?Rh₂?PPD,Rc?C-Mc1?F₂?CK?PPD;Ginsenoside Rb₂ converts to 7metabolites,with the pathway of Rb₂?Rd?F₂?CK?PPD,Rb₂?Rd?Rg₃?Rh₂?PPD,Rb₂?CO?F₂?CK?PPD,Ginsenoside Rb₃ converts to 7 metabolites,with the pathway of Rb₃?Rd?F₂?CK?PPD,Rb₃?Rd?Rg₃?Rh₂?PPD,Rb₃?Gyp-IX?F₂?CK?PPD.3.PPD-type ginsenosides conversion and transformed efficiency:When sample transformed for 60 hours,the transformed efficiency calculated as below:Ginsenoside Rb₁95.06%,Ginsenoside Rc 93.83%,Ginsenoside Rb₂ 92.71%,Ginsenoside Rb₃ 93.81%,Ginsenoside Rd 93.14%,Ginsenoside F₂ 92.48%,Ginsenoside Rg₃ 91.82%,Ginsenoside CK 13.23%,Ginsenoside Rh₂ 16.57%.4.Diversity analysis of gut microflora:In the level of phylum,the sample with ginsenoside have obviously increase on Firmicutes and Proteobacteria than the sample without;the sample with ginsenoside have obviously decrease on Bacteroidetes than the sample without?p<0.01?.In the level of genus,the sample with ginsenoside have obviously increase on Escherichia-Shigella than the sample without;the sample with ginsenoside have decreases on Dorea,Prevotella₉,Megasphaera,while there is a obviously decrease on Dorea?p<0.01?.Conclusion:Human gut microflora has metabolic effect on PPD-type ginsenosides,this effect mainly proceeds in deglycosylation,and metabolizes ginsenosides to secondary aglycones,which absorbed in blood system.During this convert process,PPD-typel ginsenosides appear to be stable in metabolic form and metabolic time,metabolic end product will be PPD,but that is not a perfectly convert,most components appear to be ginsenoside CK and ginsenoside Rh₂.PPD-type ginsenosides help increase beneficial bacterias and reduce harmful ones,by changing structure and composition in phylum and genus,this may bring positive impact to disease treatment and health care function.