Study On The Mechanism Of Effective Components In Gynura Procumbens Against Acute Kidney Injury

Acute renal injury(acute kidney injury,AKI)is a common clinical renal disease with high mortality caused by a variety of causes,which can be caused by drugs,surgery,renal ischemia and external stimulation.In recent years,with the development of medical technology,a variety of human diseases have been effectively treated,but the treatment of AKI has not been essentially improved,its medical cost is high,and the related morbidity and mortality even have a trend of increasing year by year.Recent studies have shown that incomplete repair of AKI can lead to kidney fibrosis and eventually lead to chronic kidney disease(CKD).More and more evidence shows that the occurrence of AKI and subsequent renal repair work is complex and multifactorial,involving renal tubules,microvessels and a variety of inflammatory mechanisms,the interaction of these mechanisms will magnify and aggravate the development of AKI.It is for these reasons that AKI has been widely concerned by the international community and has obviously become an urgent public health problem faced by the whole world.Gynura procumbens is a perennial herbal medicine and food homologous plant,it has been reported that it has a good anti-inflammatory effect,but so far,its active components and mechanism are not clear.Accordingly,this paper carries out the following research:1.Efficacy analysis and effective fraction screening of extract of Gynura procumbens from chrysanthemum.At present,the research on the active substances of Gynura procumbens is mainly focused on its ethanol extract,but its activity in the treatment of AKI has not been reported,and the study on its effective medicinal parts has not been reported.The effective components can be quickly separated by ethanol extraction,and the effective parts can be quickly selected by comparing the efficacy results of different medicinal parts.Therefore,in this study,glycerol-induced Wistar rats were selected to establish AKI model.By comparing the biochemical indexes and HE staining results of each group,it was found that the ethyl acetate part of the ethanol extract of Gynura procumbens had the most obvious effect.This study laid a foundation for the discovery of follow-up active components.2.Analysis of effective components and study on the efficacy of effective components in vivo and in vitro.In the previous study,we found that the ethyl acetate part of Gynura procumbens was mainly phenolic acids.Therefore,in this study,the LC-MS technique was used to infer that the main active components of ethyl acetate of Gynura procumbens were3,5-dicaffeoylquinic acid,1,3-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid.The contents of the three main components were determined by HPLC.Through the rat kidney podocytes induced by LPS,the effects of the three main active components on inflammatory cells were investigated,and the best active ingredient was 4,5-dicaffeoylquinic acid.Finally,using the AKI rat model,the levels of BUN and CRE in serum,kidney index and kidney tissue structure of rats in different administration groups were compared.It was found that 4,5-dicaffeoylquinic acid had the effect of treating AKI.This experiment laid a foundation for the follow-up study of the mechanism of treatment of AKI with 4,5-dicaffeoylquinic acid.3.Study on the Therapeutic Mechanism of 4,5-dicaffeoylquinic acid on Glycerol-induced Acute Renal injury in Rats based on Plasma metabonomics.Metabonomics is generally used as a means of disease diagnosis and drug analysis,and it has been considered as a promising and reliable new diagnostic method.The metabonomics method based on LC-MS can quickly obtain the comprehensive information of endogenous metabolites in biological samples.In this study,we combined plasma metabonomics and Western blot technology to verify the mechanism of proteins related to AMPK/mTOR signal pathway.The results of plasma metabonomics showed that 4,5-dicaffeoylquinic acid exerted its effect mainly by affecting fatty acid metabolism and pyruvate metabolism,and had significant regulatory effects on six components(3-methylhistidine,guanidoacetate, phosphatidylcholine O-38:1,phosphatidylethanolamine O-42:3,3-indoxyl Sulfate,Malic acid).The results of;Western blot showed that p53,ULK1 and Glutaminase were significantly up-regulated in AMPK/mTOR signal transduction pathway.The results showed that 4,5-dicaffeoylquinic acid exerted its effect on fatty acid metabolism and pyruvate metabolism mainly by affecting fatty acid metabolism and pyruvate metabolism.The results showed that p53,ULK1 and Glutaminase were significantly up-regulated in AMPK/mTOR signal pathway.AMPK,mTOR,IDH2,IDH1 and UCP2 decreased significantly.Therefore,4-dicaffeoylquinic acid has the efficacy of treating AKI,which lays a solid theoretical foundation for the research and development of new drugs and further research.