基于生物分析及动物实验探讨仙方活命饮治疗Ⅲ型前列腺炎作用机制OA

Objective:To explore the mechanism of Xianfang Huomingyin(XFHMY)in the treatment of type Ⅲ prostatitis(CP/CPPS)through network pharmacology,molecular docking,and animal experiments.Methods:The traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and the Swiss Target Prediction database were used to screen and sort out the active ingredients and corresponding targets of XFHMY.The potential therapeutic targets of CP/CPPS were collected from online databases,such as the Online Mendelian Inheritance in Man(OMIM),GeneCards,and DisGeNET.The potential core targets of XFHMY for treating CP/CPPS were further screened by constructing a protein-protein interaction(PPI)network and performing topological analysis.Meanwhile,the DAVID database was chosen to perform enrichment analysis on the intersection targets.On this basis,the AutoDock software was used for molecular docking,and the data was subsequently imported into the GraphPad Prism 8 software to generate a heat map.SD rats were divided into seven groups:A blank group,a sham operation group,amodel group,low-,medium-,and high-dose XFHMY groups(3.645,7.29,14.58 g·kg-1),and a tamsulosin hydrochloride group(0.018 mg·kg-1).Hematoxylin-eosin(HE)staining was used to evaluate the pathological changes in prostate tissue.The inflammatory factor indicators of rats in each group were detected via enzyme-linked immunosorbent assay(ELISA).Real-time fluorescence quantitative reverse transcription polymerase chain reaction(Real-time PCR)and Western blot were used to evaluate the mRNA and protein expression levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),and nuclear transcription factor-κB(NF-κB)p65 in prostate tissue.Results:The HE staining showed no significant signs of inflammatory cell infiltration in the prostate of the sham operation group compared to the blank group,while the model group had significantly inflammatory cell infiltration.The ELISA results showed that compared to the blank group,TNF-α,IL-1β,and COX-2 in the sham operation group had no significant differences.However,they were significantly higher in the model group(P＜0.01),indicating successful CP/CPPS modeling in rats.Compared with the model group,the low-,medium-and high-dose XFHMY group and the tamsulosin hydrochloride group showed significant decreases in TNF-α,IL-1β,and COX-2(P＜0.05,P＜0.01).The Real-time PCR analysis revealed that compared to the model group,the low-dose XFHMY group had reduced Akt and NF-κB p65 mRNA expression(P＜0.05,P＜0.01).In the medium-and high-dose XFHMY group and tamsulosin hydrochloride group,PI3K,Akt,and NF-κB p65 mRNA levels decreased significantly(P＜0.05,P＜0.01).Western blot analysis showed that compared to the model group,the low-dose XFHMY group had lower p-NF-κB p65/NF-κB p65(P＜0.05).The medium-and high-dose XFHMY group and the tamsulosin hydrochloride group showed significant decreases in p-PI3K/PI3K,p-Akt-ser473/Akt,p-Akt-thr308/Akt,and p-NF-κB p65/NF-κB p65(P＜0.01).Conclusion:XFHMY may exert therapeutic efficacy on CP/CPPS by inhibiting the PI3K/Akt/NF-κB signaling pathway and reducing inflammatory responses.Additionally,NF-κB activation may be related to the activation of ser473 and thr308 sites.