| The treatment processes and research quality of raw materials of Chinese medicine are key scientific issues for development of traditional Chinese medicine preparations and ensure its clinical efficacy.The scientific understanding of the basis of traditional Chinese medicine is important to guide the preparation of raw materials processing,and to ensure the clinical efficacy of pharmaceutical products.Moreover,the chemical composition of traditional Chinese medicine is diverse and acting on the systematic pathological network,thus showing a complex "quantity-effect"relationship.Based on the overall view and systematic nature of Chinese medicine,the task group put forward the essence of the material basis of traditional Chinese medicine,named "structural components".There is a certain amount of "quantity" and"quantity" between the components and ingredients,which curing disease based on the complex "component-target-effect" relationship.Because of component is the basic unit of traditional Chinese medicine preparation,it is important to find the excellent components of raw materials and clarify its structural characteristics,then explore the relationship between the components and disease,which will lay the foundation for the development of traditional Chinese medicine preparation.In this study,terpene glycosides/polysaccharide two-component from Moutan Cortex were analyzed on the characteristics of structural components and the regulatory mechanism according to the pathological model.Moreover,the active components were confirmed by the data mining technology of network pharmacology and the relationship between the multi-component and the pathological target.The rat model of diabetic kidney disease was established for evaluating the relationship of"component-efficacy" and confirming the efficacy of raw materials of Moutan Cortex.Based on the analysis of gene expression profiles and pathways,the synergistic relationship of different components on multiple targets and pathways was studied based on the multi-link effect,which laid the foundation for constructing a drug delivery system with component as the basic unit.In this study,the biological effect as a starting point,with clear structure of components,which provided a systematic research ideas for finding effective and stable raw materials of traditional Chinese medicine preparation.The main contents include the following sections:1.Construction of relationship of ’"ingredients-targets" based on network pharmacology and explore the active components for preventing diabetic kidney diseaseAccording to the Pubchem database,the chemical database and the TCMSP database were used to identify 31 major active compositions in the MC with OB(oral bioavailability)>20%and DL(drug-likeness)>0.18 as the screening index.By using the model of drug target prediction,the target of predicting the active ingredients was also obtained by text mining,and 130 potential targets were obtained.After normalization of the target,the GO 3.0 functional analysis system and the KEGG pathway annotation analysis were performed using the MAS 3.0 biomolecule functional annotation system.The results showed that there were 78 potential regulatory pathways of MC,which had 32 channels related to the pathogenesis of DKD,which may be mainly involved in the main biological processes such as extracellular matrix,inflammatory reaction and oxidative stress.From the analysis of network of the component-target-biological process,we found that each active ingredient often regulates multiple targets,and the target proteins often share ligands,showing the network of biological systems.The molecular mechanism of MC for prevention DKD showed as the multi-component,multi-target and multi-channel.In addition,the analysis of network topology suggested that the terpene glycosides from MC(MC-TG)may be the main effective component of MC.2.Analysis of the structural components of raw materials from Moutan Cortex for prevention diabetic kidney diseaseThe terpene glycosides(MC-TG)and polysaccharide(MC-P)were the active components of MC.In this chapter,the preparation of the components was carried out to lay the foundation for the mechanism research of multi-component.The results showed that the yield of MC-TG was 3.67%.The content of terpene glycosides was 45.5%,The structural proportions of ingredients in the component are:paeoniflorin:paeoniflorin:benzoyl paeoniflorin = 3.41:2.48:1.00.Secondly,the MC-P was purified by DEAE-52 ion exchange chromatography column.The polysaccharides of MC-Pa and MC-Pb were obtained and their physicochemical properties,structural analysis and monosaccharide composition were studied.The results showed that the yield of MC-Pa:MC-Pb was 3.18:1.63,the glucose content of MC-Pa was 86.5%,the protein content was 1.67%,the composition of monosaccharide was as follows:Mannose:L-rhamnose:galacturonic acid:D-glucose:L-arabinose:xylose=1.00:2.25:3.21:5.89:2.52:6.22;the glucose content of MC-Pb was 92.4%,the protein content was 1.08%,the composition of monosaccharide was as follows:Mannose:L-rhamno se:galacturonic acid:D-glucose:L-arabinose:xylose=1.00:0.71:2.75:0.19:0.69:1.95;IR and UV analysis showed that MC-Pa and MC-Pb had the characteristic absorption spectra of polysaccharides.3.Evaluation of the relationship of "component-efficacy" of terpene glycosides/polysaccharide two-component from Moutan Cortex based on the pathogenesis of renal injuryIn this study,we established the DKD rat model to evaluate the efficacy of the two-component of MC on improving the renal injury of DKD rats by using the renal function parameters,renal histopathological examination and the cytokines which closely related to the pathogenesis of DKD.The results showed that the two-component of MC was able to reduce the content of blood glucose,serum creatinine,blood urea nitrogen and total protein in the kidney;improve the glomerular basement membrane thickening,interstitial inflammatory cell infiltration,podocyte structure damage;inhibit the protein expression of FN and Col IV in renal tissue;improve the activity of T-SOD and GSH-PX in serum;and control the levels of VCAM-1 and IL-1β in the kidney and serum of rats.This study suggested that the two-component of MC could significantly improve the renal injury of DKD rats,the effect is better than MC-TG or MC-P alone administration.In addition,this effect can be equivalent to the total extract of MC.Moreover,the active components could regulate different biological process and had synergistic effects.4.Advances in the effect of terpene glycosides/polysaccharide two-component from Moutan Cortex on diabetic kidney disease based on the integrated regulatory mechanism of "structural components-biological network"(1)The gene expression profile of each group was analyzed by cDNA microarray,and the effect of various components on the regulation of biological system network was discussed at the level of mRNA,which provided reference on the regulatory mechanism of the multi-component from MC in curing DKD.The results showed that the number of different expressed genes was the highest among the model group and the blank group,and the number of different expressed genes was up to 859,the number of genes was down to 652,and the biological network of the rats was speculated.Compared with the model group,the number of up-regulated genes was 808,and the number of down-regulated genes was 537,which indicated that it could regulate the expression of multiple genes and had a strong regulatory effect on biological network.Compared with the blank group,the number of different expressed genes was much smaller than compared with the model group,suggesting that the two-component of MC could regulate network to normal.In addition,based on the analysis of the function of different genes,we known that the multi-component of MC can be involved in the biological process of drug reaction,inflammatory reaction,oxidative stress and so on;and MC-TG may be mainly involved in biochemical reactions,drug reactions,inflammatory response and other biological processes;MC-P components may be mainly involved in the biological process of biochemical reactions,redox reactions,oxidative stress and so on.It could be seen that multiple components of MC could intervene DKD pathological network by participating in multiple biological processes,and each component had the emphasis of regulating biological process,and had the characteristics of synergistic and multi-link regulation.(2)Based on the analysis of the target and pathway of MC,the efficacy evaluation and the result of cDNA microarray,the effect of different components of MC on the control of different pharmacological links was carried out.We found that MC-TG was able to regulate the expression of TGF-β1/p38MAPK to inhibit extracellular matrix deposition.MC-TG could significantly down-regulate IRE1a and NF-κB p65 in the regulation of mesangial to inhibit ERS induced inflammatory response.In addition,MG-P could significantly scavenge free radicals in vitro and in vivo and inhibit the ROS production induced by AGEs in terms of antioxidant stress.Irn conclusion,the two-component of MC can regulate the development of DKD by regulating multiple pathological links based on the integrated regulatory mechanism of "structural components-biological network". |
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