| Ginger is the dried rhizome of Zingiber officinale Rosc,Which has the effects of assist yang and disperse cold,restore Yang and rescuing patient from collapse,warm the lung and dissolve fluid retention,and is used clinically to treat vomiting,diarrhea and anti-inflammatory.The processed ginger is made of dried ginger by sand scalding.It has the functions of warming channel for arresting bleeding and deficiency-cold in the spleen and stomach.It is commonly used in clinic to stop bleeding,treat abdominal pain,diarrhea and anti-inflammatory.Ulcerative colitis is a common digestive system disease with diarrhea,blood in the stool,and intestinal inflammation as the main symptoms.Due to it's complicated pathogenesis,the progress of finding effective prevention and treatment drugs has been limited in recent years.Dried ginger and processed ginger has potential value in the treatment of ulcerative colitis.In this study,the intervention of alcohol extract and crude polysaccharide of dried ginger and processed ginger on dextran sodium sulfate induced ulcerative colitis in mouse was studied by metabonomics,in order to provide a new evidence for the treatment of clinical ulcerative colitis.Methods:(1)According to the fourth part of The Chinese Pharmacopoeia(2015 Edition),made the processed ginger.Dry ginger and processed ginger pieces were extracted by water and ethanol respectively to make crude polysaccharide;Then 80% ethanol was used for reflux extraction and spin-drying to prepare alcohol extracts.Establishment of ulcerative colitis mouse model group by freely drinking 2% dextran sodium sulfate and give a certain volume of normal saline by gavage every day.Then,the dried ginger and processed ginger's polysaccharide administration groups(400 mg / Kg)and alcohol extraction administration groups(200 mg / Kg)were respectively administered to the model mouse in seven days,the blank group ate and drank freely and were given a certain volume of normal saline by gavage every day,then all mice were sacrificed on day 8.By observing the changes of body weight,disease activity index(DAI),spleen index,pathological section of colon tissue and the content of inflammatory factors,the anti-inflammatory effects of alcohol extract and crude polysaccharide of the dried ginger and processed ginger were evaluated.(2)UHPLC-HRMS was used to detect metabolites in the blank and model serum of normal mice and ulcerative colitis mice.Partial least squares discrimination(PLS-DA)was used for multivariate analysis to screen for differential metabolites.The differential metabolites were introduced into MetaboAnalyst to build metabolism pathway for analysis of metabolites and metabolic pathways related to ulcerative colitis.(3)Targeted metabolomics was used to analyze the changes in bile acid spectrum in the mouse liver and colon contents of normal group,ulcerative colitis mouse group,and each administration group.The related pathways and metabolism involved in bile acids were analyzed by consulting relevant literature.Then to explore the mechanism,similarities and differences of the effect of each administration group on ulcerative colitis from the overall metabolic level.(4)Based on the mass spectrum analysis method of UHPLC-HRMS to investigate the differences in chemical composition between the methanol extracts of dried ginger and processed ginger,and related compounds were identified based on precise molecular mass,ion fragmentation rules,retention time,etc.Multivariate analysis using PLS-DA to screen out different chemical components.Combined with the in vitro anti-inflammatory(NO)experiment using Raw 264.7 mouse macrophages,the reasons for the changes in pharmacological efficacy of dried ginger after processing were analyzed.Results:(1)Compared with the blank group,the weight of the model group was significantly reduced(P<0.05),the colon length was significantly shortened(P<0.05),the disease activity index,spleen index,TNF-?,IL-1 ?,and IL-6 were significantly increased(P<0.05).The results showed that the model group was successful.After administration,it was found that compared with the model group,the weight loss,disease activity index,colon atrophy and the increase of three inflammatory factors in each group were significantly alleviated(P<0.05).(2)Based on UHPLC-HRMS metabonomics and pathway analysis,18 differential metabolites were identified from serum,which mainly involved bile acid synthesis pathway,taurine and taurine metabolism pathway.(3)Based on targeted metabolomics analysis,31 bile acids were found from the liver,and 50 bile acids were found in the colon contents.Multivariate analysis showed that the relative content of 12 bile acids in the liver and 26 bile acids in the colon contents changed significantly.Metabolic pathway analysis showed that the abnormal metabolism of bile acid may be related to the abnormal function of farneside receptor,G protein coupled receptor and vitamin D receptor in inflammatory state.At the same time,we found that secondary bile acids may play a more important role in regulating ulcerative colitis,and compared with other drug-administered groups,the alcohol extract group of processed gingercould more effectively regulate secondary bile acid metabolism disorders.(4)A total of 74 compounds including gingerols,diphenylheptanes and flavonoids were identified by mass spectrometry based on UHPLC-HRMS,and the contents of 11 gingerols ? shogaols and zingerones were significantly increased after processing.Combined with Raw 264.7 mouse macrophage anti-inflammatory(NO)in vitro experiments showed that the anti-inflammatory effect of shogaols is better.Conclusion:In this study,it was found that the ethanol extracts and crude polysaccharide active substances of dried ginger and processed ginger could effectively alleviate the pathological condition of mouse with ulcerative colitis.And bile acid is closely related to ulcerative colitis.The abnormal metabolism of bile acid may be related to the abnormal function of farneside receptor,G protein coupled receptor and vitamin D receptor in inflammatory state.Among them,secondary bile acids may play a more important role in regulating ulcerative colitis,and compared with other drug-administered groups,the alcohol extract group of processed ginger can more effectively regulate secondary bile acid metabolism disorders.At the same time,it has been preliminarily proved that zingiberol compounds may be the material basis for different functions of dried ginger and processed ginger. |
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