Based On The "Abnormal Ascent-descent Between Qing And Zhuo" Theory To Study The Liquid Biopsy Marker Of Urinary Exosomal RNA In Renal Interstitium Fibrosis

BackgroundRenal interstitial fibrosis(RIF)belong to the traditional Chinese medicine "edema","asthenic overstrain","lumbago","turbid urine","Long Bi" and "off grid".The disease has long duration and it is hard to diagnose and treat.In the long-term clinical observation and treatment,traditional Chinese medicine has formed a unique theory of TCM pathogenesis,which is used to guide the treatment of renal interstitial fibrosis by TCM with reliable effect.With the development of modern natural science and technology,the diagnosis and detection of renal interstitial fibrosis under the guidance of pathogenesis theory will also be developed and popularized.ObjectiveTo sort on “qing zhuo” and “abnormal ascent-descent between clear and turbid” theory,and the relationship with RIF,the liquid biopsy index of renal interstitial fibrosis urinary exosomal are studied under the guidance of the traditional Chinese medicine theory "voicing coherent",for further explanation and enrichment the theoretical connotation and material basis of "voicing coherent" and providing laboratory gauge for monitoring and guiding the treatment of "ascending and descending turbidity method".Methods1.Theoretical research: To explore "voicing" and "voicing coherent" on the historical and current literature,and analyze the relationship of "voicing coherent" and renal interstitial fibrosis,urinary exosomes.2.Experimental studies under the guidance of "voicing coherent" pathogenesis theory includes:(1)The establishment of experimental methods: Urinary exosomes are enriched by polyethylene glycol precipitator(PCP),after verified by electron microscope and western blot,qRT-PCR method are used to monitor the housekeeping gene of miRNA(U6)and mRNA(β-actin).The result is to establish the basic technology of research.(2)urinary exosomes are enriched by PCP.After identification,qRT-PCR method is used to detect exosomal miR-29 c,miR-21,E-cadherin and vimentin in urine.Results1.The “qing” and “zhuo” are the "classic" concept,the meaning is rich,can be used to represent a useful / "the subtle" and "dross" /harmful substances.“Separating clear and excreting turbid” happen in the healthy and physiological body and "Voicing coherent",is the voicing of the disorder,belongs to the pathology of disease states.Epithelial mesenchymal transition is one of the important mechanisms of renal interstitial fibrosis,the expression disorder of renal tubular epithelial source markers(E-cadherin,miR-29c)and interstitial source markers(vimentin,miR-21)in the process of renal interstitial fibrosis epithelial mesenchymal transition may be a molecular mechanism of “voicing coherent ".2.Urinary exosomes could be enriched successfully by PCP.The exosomes enriched by this method could be used for the subsequent detection of the cargo protein and nucleic acid.The experimental operation is simple and the price is cheap,which lays the foundation for the development of the following index of liquid biopsy.3.The detection of urinary exosomal RNA in patients with renal interstitial fibrosis confirmed that urinary exosomal E-cadherin,miR-29 c and vimentin and miR-21 all changed to some extent(or disorder).The exosomal E-cadherin,miR-29 c and vimentin,miR-21 in urine may conform to "voicing coherent".4.The expression of exosomal miR-29 c in patients with renal interstitial fibrosis are reduced and the results are stable,and decreased more significantly in the trend with the degree of fibrosis severity,is related to blood urea,creatinine,cystatin C and eGFR,suggesting that it is a relatively stable marker in the degree of renal interstitial fibrosis "voicing coherent".ConclusionsmiR-21 and vimentin which are auxo-action on RIF have an increasing tendency in the urinary exosomes,E-cadherin and miR-29 c,which have inhibitory effects on RIF,this agree with the renal interstitial fibrosis pathogenesis "voicing coherent",and can reflect the molecular mechanism of RIF "abnormal ascent-descent between qing and zhuo" for a certain degree.Of which the urinary exosomal miR-29 c is stable expression in urinary exosomes and reduce with fibrosis degree,and may be a relative stability marker of the "abnormal ascent-descent between qing and zhuo".