Action Mechanism Of Modified YinChen ShiXiao Powder In The Treatment Of Chronic Renal Failure Based On Network Pharmacology And Animal Experiment

Objectives:Chronic renal failure(CRF),a kind of disease with high incidence,is the final-stage kidney disease seriously endangering people’s health resulting from the development of various chronic kidney diseases.Modified Yin Chen Shi Xiao Powder,which Professor Wang Yaoguang often adopts to treat the chronic renal failure clinically,can effectively delay the development of the disease.Based on the method of network pharmacology,this thesis explores the potential targets and pathways of Modified Yin Chen Shi Xiao Powder in the treatment of CRF,so as to verify the effects of different concentrations of Modified Yin Chen Shi Xiao Powder on the rats with adenine-induced CRF and related pathway mechanisms through animal experiments,and provide a scientific basis for the clinical treatment of CRF as well as the promotion and application of this prescription.Methods:1.Screen out the active compounds of Modified Yin Chen Shi Xiao Powder and the potential targets for the CRF treatment based on the network pharmacology,conduct the GO(gene ontology)enrichment analysis and the Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis on the intersecting target genes,and construct the compound-target network and the protein-protein interactions(PPIs).2.Create the rat model of adenine renal failure,divide 60 health-level Wistar rats into groups.Except for the control group,other groups were fed with losartan potassium and different concentrations of Modified Yin Chen Shi Xiao Powder decoction for 14 days,then compared the model groups with the control group for analysis.Observed the general condition of the rats and their kidneys,the weights and the ratio of kidney weight/body weight,then observed the pathological changes of renal tissue through HE staining and Masson staining,compared the serum scr,BUN and 24 h urine protein of the rats.3.According to the results of network pharmacology,the PI3K/Akt/m TOR pathway was selected as the research object,and the Western blot and q RT-PCR technology was adopted to detect the changes of the pathway index,so as to verify the effect of Modified Yin Chen Shi Xiao Powder on the pathway and relevant proteins.Results:1.By searching 8 herbal ingredients(Astragalus Flavone,largehead atractylodes rhizome,capillary wormwood herb,trogopterus dung,longbract cattail pollen,rhubarb,oyster,glabrous greenbrier rhizome)of Modified Yin Chen Shi Xiao Powder through the database,a total of 85 chemical components are obtained,of which 64 such as quercetin,jaranol,kaempferol,testosterone palmitate and naringenin are related to chronic renal failure;a total of 219 targets are involved,of which the MAPK3,MAPK1,JUN,RELA,SRC,TP53,AKT1 shows the greatest correlation.Moreover,a total of 3,003 biological processes,123 cell components,270 molecular functions and 191 related pathways are detected to be enriched into the treatment of chronic renal failure.Some of the signaling pathways have relatively obvious effects such as the atherosclerosis-related pathways,PI3K/Akt signaling pathways,AGE-RAGE signaling pathways and cancer-related pathways.2.(1)2 rats fall out during the feeding and experiment.The initial weights of six groups of rats are normal with statistical difference(P>0.05),the mental conditions are good and the hairs are glossy;after 21 days of adenine molding,the weights in 6 groups of rats have increased,and the weight of the control group is significantly higher than that of other groups(P<0.01).After 14 days of administration,the weight of other five groups of rats is still lower than that of the control group(P<0.05),the weight of rats in the medium herbal dose group is slightly higher than that of the model group(P<0.05),and the weight of rats in the high herbal dose group and losartan potassium group is significantly higher than that of the model group(P<0.01).The renal coefficients of other groups are also higher than those of the control group: compared with the model group,the losartan potassium group has a relatively low renal coefficient with the medium herbal dose group(P<0.05),while the weight of high herbal dose group is significantly lower than that of the model group(P<0.01).(2)Pathological staining results show no abnormalities in the glomerular,tubular,renal interstitial and cell morphology of rats in the control group,except for a small amount of collagen fibers are deposited.The brown crystalline deposits appear in the other groups,and the model group shows the most serious lesion such as the changes in glomerular morphology,the cystic cavity enlargement,the uneven thickness,atrophy and partial necrosis of tubular walls,renal interstitial hyperplasia,the existence of many collagen fibers and infiltration of inflammatory cells.After 14 days of administration,the rats in model group show less histopathological changes than those in the high herbal dose group,and the high herbal dose group shows the mildest pathological changes.(3)After 14 days of administration,the serum Scr,BUN and 24 h urine protein of the rats in the model group,losartan potassium group,low herbal dose group,medium herbal dose group and high herbal dose group are higher than those in the control group,so the difference is of statistical significance(P<0.05).Except for the low herbal dose group,the indicators of other groups have been greatly improved compared with the model group,especially the high herbal dose group and the losartan potassium group have the most obvious improvement(P<0.001).3.(1)According to the Western blot results,the expression of PI3 K,AKT,m TOR,p-PI3 K and p-AKT protein in the model group is significantly higher than that of the control group,so the difference is of statistical significance(P<0.01).The expression of PI3 K and AKT protein in the low herbal dose group is lower than that of the model group(P<0.05).The expression of PI3 K,AKT and p-AKT protein in the medium herbal dose group is lower than that of the model group(P<0.05).The expression of PI3 K,AKT,m TOR,p-PI3 K and p-AKT protein in the high herbal dose group is lower than that of the model group(P<0.05).(2)According to the q RT-PCR results,the m RNA level of PI3 K,AKT and m TOR in the renal tissue of the model group has been significantly increased(P<0.01).Compared with the model group,the m RNA expression of PI3 K and AKT in the losartan potassium group has been decreased(P<0.05).The PI3 K,AKT and m TOR level of the herbal dose groups has been improved compared with that of the model group(P<0.01).The high herbal dose group shows the most obvious effect,which indicates that Modified Yin Chen Shi Xiao Powder can effectively inhibit the expression of PI3K/AKT/m TOR signaling pathway in the rats of the model group.Conclusion:1.According to the network pharmacology,Modified Yin Chen Shi Xiao Powder can play the role in the treatment of chronic renal failure through multi-component,multi-target and multi-pathway.The main ingredients such as quercetin,Jaranol,kaempferol,Testosterone palmitate,naringenin can regulate the biological processes such as response to molecule of bacterial origin,response to lipopolysaccharide,wound healing,cellular response to chemical stress,response to metal ion and cellular response to oxidative stress,so as to intervene in the atherosclerosis and cancer-related pathways as well as multiple signaling pathways including PI3K/Akt pathway and AGE-RAGE pathway.2.Modified Yin Chen Shi Xiao Powder can delay the further development of blood creatinine and urea nitrogen levels in the rats with adenine chronic renal failure,so as to effectively reduce the urine protein,delay kidney damages.The high dose group shows more obvious effect than that of other dose groups.3.Modified Yin Chen Shi Xiao Powder can regulate the expression of relevant genes and proteins in the PI3K/AKT/m TOR pathway,so it’s proved that the delayed renal failure may be related to the regulation of the PI3K/AKT/m TOR pathway.