| Objective: Data mining was used to analyze the characteristics of TCM for the treatment of jaundice,and to provide theoretical guidance and new ideas for the clinical treatment of jaundice in traditional Chinese medicine.To explore the biological mechanism of SLD(Huanglian-Huhuanglian)in the treatment of jaundice by using network pharmacology,LC-MS analysis and miRNA sequencing technology,and to provide a research basis for the pharmacological mechanism of SLD in the treatment of jaundice.Methods:1.Based on "Chinese Medical Dictionary" and "Prescription Dictionary",the prescriptions for the treatment of jaundice were screened and classified by age,and their medication rules were analyzed through data mining.2.The pharmacological mechanism of SLD in the treatment of jaundice was analyzed by network pharmacology.The active components of SLD were retrieved and screened by TCMSP database,and the corresponding target names were retrieved in Uniprot database according to "MOL ID".Disease targets related to jaundice were searched in the Gene Cards database using the keyword "jaundice".Venny2.0 software was used to obtain the common targets of "drug-disease",and Cytoscape 3.7.2 was used to draw the association network map.Finally,the key targets were analyzed by GO enrichment analysis and KEGG pathway enrichment analysis.3.Analysis the major ingredients of SLD by LC-MS.4.Establish a mouse jaundice model induced by ANIT to verify the efficacy of SLD in the treatment of jaundice.5.miRNA sequencing was performed to screen the differential mirnas and target genes of SLD in the treatment of jaundice,and GO and KEGG analysis was performed to explore the molecular mechanism of SLD in the treatment of jaundice.Results:1.Through the analysis of the prescriptions for the treatment of jaundice in different ages,it was found that there were 11 prescriptions for the treatment of jaundice in the Eastern Han Dynasty.The most frequently used traditional Chinese medicines were rhubarb,gardenia,and licorice.Gardenia,Gardenia-Phellodendron;there are 8 prescriptions for the treatment of jaundice in the Eastern Jin Dynasty,and the most frequently used traditional Chinese medicines are Yinchen,Gardenia,and light Douchi,and the Yinchen-Gardenia drug pair was screened out;There are 34 prescriptions in total,and the most frequently used traditional Chinese medicines are rhubarb,tuckahoe,and skullcap.The rhubarb-coptis,rhubarb-yinchen-gardenia,yinchen-gardenia-rhubarb drug pairs were screened out;there are 160 prescriptions for the treatment of jaundice in the Song Dynasty.First,the most frequently used traditional Chinese medicines were Yinchen,Rhubarb,and Zhigancao,and the pairs of Rhubarb-Scutellaria-Gardenia,Poria-Atractylodes were screened out;there were 11 prescriptions for the treatment of jaundice in the Jin and Yuan Dynasties,and the most frequently used traditional Chinese medicines were: Atractylodes,tangerine peel,and licorice,the Poria-Atractylodes pair was screened out;there were 161 prescriptions for the treatment of jaundice in the Ming Dynasty,and the most frequently used traditional Chinese medicines were Yinchen,Licorice,and Atractylodes.There were 160 prescriptions for the treatment of jaundice in the Qing Dynasty,and the most frequently used Chinese medicines were Yinchen,Poria,and Atractylodes.After data mining,we found SLD,a prescription for treating jaundice by clear hot-humid and heat of weakness.2.Network pharmacology analysis found that SLD has 24 active ingredients,225 drug targets,and 3456 jaundice disease targets.There are 122 intersecting targets,and 23 core targets are obtained through PPI protein network interaction screening,which are PTEN,HSP90AA1,JUN,ESR1,MYC,EGFR,PPARG,MMP9,TP53,RELA,IL6,CASP3,HIF1 A,TNF,CCND1,PTGS2,CCL2,FOS,MMP2,NFKBIA,AKT1,CXCL8,IL10.Involved in reactive oxygen species metabolic processes,cellular responses to chemical stress,lipopolysaccharide responses,responses to molecules of bacterial origin,oxidative stress;membrane rafts,membrane microdomains;biological functions such as DNAbinding transcription factor binding,lipids and atherosclerosis sclerosis,hepatitis B,Kaposi’s sarcoma-associated herpesvirus infection,human cytomegalovirus infection and other signaling pathways.3.The results of LC-MS showed that the major ingredients of SLD may be quercetin,berberine,catechin,albiflorin.4.The results of miRNA sequencing and q-PCR verification showed that SLD treated jaundice mainly by upregulating mmu-miR-30b-3p,mmu-miR-486b-5p,mmu-miR-146a-5p,and downregulating mmu-miR-465a-3p.Differences before and after treatment miRNAs were mainly enriched in cell parts,organelles,binding,catalytic activity,cellular processes and other biological processes,signal transmission,viral infectious diseases,transport and metabolism,folding/sorting/degradation(PI3K/Akt signal pathways),lipid metabolism and other signal pathways.5.After target gene screening and,it was found that Rhov,Ripk1,Arhgef18,Nadsyn1,Baiap2l1,Vmn2r110,PTEN,Enox1 and MMP2 were the target genes of differential miRNA regulated by SLD in the treatment of jaundice.Through further q-PCR verification,it was found that SLD could decrease the expression of PTEN and MMP2 of jaundice mice.Conclusions:1.Shuanglian Dan is a little prescription for jaundice treatment,which embodied "clearing dampness-heat and deficiency heat,and combined treatment of liver and gallbladder,spleen and stomach".2.The main components of SLD were berberine and quercetin.3.The main active components of SLD in the treatment of jaundice might be PTEN,FOS,AKT1.4.In this study,four potential miRNA targets for the treatment of jaundice were preliminarily identified and verified: mmu-miR-30b-3p,mmu-miR-486b-5p,mmu-miR-146a-5p,and mmu-miR-465a-3p.5.Downstream regulated genes of potential miRNA targets for SLD treatment of jaundice include MMP2,PTEN,Rhov,Ripk1,Arhgef18,Nadsyn1,etc.6.The treatment of jaundice by SLD may involve oxidative stress process and PI3K/AKT signaling pathway. |
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