Effect Of Danzhi Qing’e Formula On Benign Prostatic Hyperplasia And Urination Function In Rats

Objective:Benign prostatic hyperplasia(BPH)is a common urinary system disease with high incidence in elder men,which is accompanied by a series of lower urinary tract symptoms(LUTS).The incidence of renal qi failure increases with age,which seriously affects the quality of life.Danzhi Qing’e formula(DZQE)is composed of Psoralea corylifolia Linn.,Eucommia ulmoides Oliver,Salvia miltiorrhiza Bunge and Anemarrhena asphodeloides Bunge,which has the functions of tonifying kidney yang,promoting blood circulation and removing blood stasis,clearing heat and nourishing yin.The aims of our study were to investigate the effect of Danzhi Qing’e formula on inhibition of BPH and its related LUTS,and further uncover the potential mechanism involved.Methods:Male Wistar rats were castrated and subcutaneously injected with oestrodial/testosterone(1:100)to establish the BPH model.The effects of extract of DZQE(1.35 g/kg,2.7 g/kg,5.4 g/kg),)on the progression of BPH in rats were observed after 28 days.The bladder detrusor muscle strips of normal rats were isolated and the relaxing and contractile effects of DZQE on basal and KCl-induced detrusor strips were detected.The bladder perfusion was used to detect the effect of DZQE formula on urinary parameters in rats.Based on network pharmacology to predict the potential targets of DZQE in inhibition of BPH,samples from animal and cells(human benign prostatic hyperplasia cells(BPH-1)and prostate stromal cells(WPMY-1)were used to verify the candidate targets predicted by network pharmacology.Results:Compared with the sham-operated(Sham)group,the prostate weight and PI of in BPH group were significantly increased.After 28 days of continuous administration,DZQE at the concentrations of 1.35 g/kg,2.7 g/kg and 5.4 g/kg significantly inhibited PI in BPH rats.Pathomorphological analysis showed that DZQE significantly alleviated the disorder of prostatic epithelium and reduced the hyperplasia of prostatic stromal components in BPH rats.DZQE also significantly increased the level of testosterone(T)in serum,decreased the ratio of 17β-estradiol/testosterone(E2/T),and decreased the level of dihydrotestosterone(DHT)in serum,both of which were highly increased in BPH rats.Strip assays showed that DZQE promoted bladder detrusor contraction,increased detrusor tension,but inhibited KCl-induced detrusor contraction.Furthermore,DZQE significantly reduced the up-regulation of detrusor leakage point pressure(DLPP)and maximum voiding pressure(MVP)in BPH rats.By network pharmacological screening,102 active components and 258 potential predictive targets of DZQE were obtained,and 642 targets of BPH-related diseases were set,which were further overlapped and 86 common targets with 92 active components of BPH in the formula were finally confirmed.GO functional enrichment analysis showed that the mechanism of DZQE on inhibition of BPH involved biological processes such as cell proliferation and apoptosis,estrogen response,MAPK cascade,protein phosphorylation,aging,vasoconstriction,smooth muscle contraction and so on.KEGG enrichment results showed that DZQE inhibited BPH through 98 signaling pathways,including estrogen,MAPK,HIF,ErbB,hepatitis B,PI3K-Akt,prostate cancer and so on.The CCK8 assay showed that DZQE inhibited the proliferation of BPH-1 cells and WPMY-1 cells.Western blotting assay showed that DZQE promoted prostate ERβ expression,inhibited the expression of Aromatase in BPH-1 cells,but had no effect on the expression of ERα.And also,can inhibit the activation of ERK promote and promote the expression of p38 in MAPK pathway.Conclusion:Danzhi Qing’e formula inhibited BPH and regulate urination function in rats though estrogen and MAPK signaling pathways.