Mechanism Research On Benign Prostatic Hyperplasia Intervened By Danggui-Beimu-Kushen Pill Based On Label Free Proteomics

Objective:The intervention of Danggui-beimu-kushen Pill on Benign Prost atic Hyperplasia(BPH)has been systematically investigated using label free proteomics techniques.The possible treatment mechanisms will be reported throughthe screening of differentially expressed proteinsMethods:32 SD male rats will be used for the experiment.8 of them were randomly selected as normal control group(NC).The rest 24 rats were modeled for medicine intervention.In details,these rats were surgically castrated first.After the surgery,testosterone propionate was injected once per day for 21 continuous days with a dosage of 4mg/(kg·d).NC was sham operated then injected with abacteria olive oil for 21 days using the same dosage.After that,the 24 modeled rats were randomly split into three groups,namely,model group(M),Danggui-beimu-kushen Pill group(DBK),and positive control group(PC)Each group has 8 rats equally.Next,NC and M rats received physiological saline(1ml/100g)by gastric lavage daily.DBK and PC rats received Danggui-beimu-kushen Pill(lml/100g)and finasteride mixed suspension(1ml/100g)by gast ric lavage daily,respectively.All rats were sacrificed after 21 days medicine treatment.The prostate tissue and the serum of rats were collected for experimental analysis,such as measuring the prostate wet weight,calculating the prostatic index,observing HE staining changes of prostate pathological using optical microscopy,and testing the Testosterone(T),Estradiol(E2)andDihydrotestosterone(DHT)in serum by radioimmunoassay technique.The differentially expressed protein of prostate tissue from each rats group was evaluatedusing label free prot eomics technique,then screened if the fold change is>1.2(<0.83)and P<0.05,hence analyzed using Gene Ontology(GO)annotation andenrichment and Kyoto Encyclopedia of Gene and Genomes(KEGG)annotation and enrichment,and Protein-protein interaction network analysis(PPI).Results:①The prostate wet weight and PI of rats from M are significantly increased than those of NC(P<0.05).Compare with the samples from M,the prostate wet weight and PI of rats from DBK and PC are significantly decreased(P<0.05).②The results of histopathology examination reveal that the prostate hyperplasia is apparently observed in M.Furthermore,the prostrate hyperplasia of the DBK and PC is significantly alleviated or even cured as compare to M.③The test of serum sex hormones shows that both T and DHT are significantly increased(P<0.05)in M as compare to NC.On the contrary,E2 is significantly decreased(P<0.05).Compare with the samples from M,both T and DHT of the DBK and PC are significantly decreased than M(P<0.05).④1334 differentially expressed proteins of BPH was screened(567 of them are up-regulated and the rest 767 are down-regulated),and 159 differentially expressed proteins was screened for Danggui-Beimu-Kushen Pill intervened BPH(116 and 43 are up-regulated and down-regulated,respectively).The Venn diagram analys is shows 88 differentially expressed proteins are same for BPH and Danggui-Beimu-Kushen Pill intervened BPH.With the intervention of Danggui-Beimu-Kushen Pill,7 proteins exhibit retracement trend,they are Gclm,Kngl,Ap3ml,Nrd1,Pitpnb,Atp5d and Tapbp.The analysis using GO annotation and enrichment shows that the differentially expressed proteins of BPH and Danggui-beimu-kushen Pill intervened BPH are mainly distributed in the cell.The main molecular functions of the differentially expressed proteins of BPH are embodied in the binding activity and the enzyme activity,hence participating various metabo lism and biosynthetic process.The main molecular functions of differentially expressed proteins of Danggui-beimu-kushen Pill intervened BHP are related to the binding activity,then involving in the metabolism process.The analysis using KEGG enrichment compares the target passway in which the differentially expressed proteins of BPH and Danggui-beimu-kushen Pill intervened BPH were in volved,suggesting the mechanism of Danggui-beimu-kushen Pill intervened BPH is associated with the regulation of focal adhesion pathway.The PPI analysis of the differentially expressed proteins involved biology function and pathway is well agreed by other analyses such as GO and KEGG annotation and enrichment.Conclusion:①Danggui-beimu-kushen Pill demonstrates its effects in alleviating the BPH of rats.The prostate wet weight as well as the PI of BPH rats are significantly decreased with Danggui-beimu-kushen treatment,same as both T and DHT values.②The mechanism of Danggui-beimu-kushen Pill intervened BPH is associated with the focal adhesion pathway and differentially expressed proteins such as Gclm,Kngl,Ap3ml,Nrd1,Pitpnb,Atp5d and Tapbp.