基于PI3K/AKT/mTOR信号通路研究黄莪胶囊抗良性前列腺增生的作用及机制

代琴慧; 胡江宁; 姚建标; 沈晨君; 赵华军; 朱智慧

doi:10.13748/j.cnki.issn1007-7693.20243105

基于PI3K/AKT/mTOR信号通路研究黄莪胶囊抗良性前列腺增生的作用及机制

Effect and Mechanism of Anti-benign Prostatic Hyperplasia of Huang’e Capsule Based on PI3K/AKT/mTOR Signaling Pathway

摘要

摘要:
目的基于网络药理学方法探讨黄莪胶囊通过磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase，PI3K)/蛋白激酶B(protein kinase B，AKT)/mTOR信号通路抗良性前列腺增生(benign prostatic hyperplasia，BPH)的作用及其机制。
方法构建丙酸睾酮诱导BPH大鼠模型，观察大鼠体质量、前列腺湿重、前列腺指数，评价黄莪胶囊对BPH的抑制作用；苏木素-伊红染色试验观察大鼠前列腺腺腔，检测各组大鼠上皮细胞厚度、腺腔面积及乳头突起的长度；ELISA检测大鼠血清中前列腺特异性抗原(prostate specific antigen，PSA)的表达；借助TCMSP、TCMID数据库筛选黄莪胶囊主要化学成分和潜在靶点；整合OMIM、GeneCards、DisGeNET数据库获得BPH相关靶点；利用Cytoscape 3.7.2构建蛋白相互作用网络；采用Metascape数据库进行GO和KEGG通路富集分析；通过AutoDock软件进行分子对接，并用Pymol进行结果可视化；免疫印记法验证黄莪胶囊抗前列腺增生KEGG筛选的作用机制。
结果黄莪胶囊降低BPH大鼠前列腺湿重、指数，显著改善前列腺组织的形态，使前列腺的上皮细胞厚度减少、乳头突起减少、腺腔面积减小；黄莪胶囊降低了BPH模型大鼠PSA的表达。筛选得到黄莪胶囊85个有效药效成分，与BPH相关的137个潜在有效靶点；相关作用靶点为TP53、AKT1、ESR1、CASP3、JUN、MYC、TNF、IL6；治疗BPH有效成分槲皮素、山柰酚、木犀草素、β-谷甾醇；富集分析分别得到GO功能2193条，KEGG富集分析195条，其抗BPH关键信号通路主要涉及癌症通路、PI3K/AKT信号通路、脂质与动脉粥样硬化、乙型肝炎、人巨细胞感染、疱疹病毒感染等。通过对前列腺组织进行蛋白表达验证发现黄莪胶囊能降低其PI3K/AKT/mTOR信号通路相关蛋白的表达。
结论黄莪胶囊通过抑制PI3K/AKT/mTOR信号通路改善BPH。

Abstract:
OBJECTIVE To investigate the anti-benign prostatic hyperplasia(BPH) effect of Huang’e capsule and its mechanism through the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mTOR signalling pathway used network-based pharmacological approach.
METHODS Testosterone propionate-induced BPH was established as a rat model. The body weight, prostate wet weight, and prostate index of the rats were measured to evaluate the inhibitory effect of the capsule on BPH. HE staining test was performed to observe the glandular cavity of the rat prostate, and the thickness of the epithelium, glandular cavity area, and length of the papillae were detected in each group. ELISA was performed to examine the expression of prostate-specific antigens in the sera of rats, and TCMSP and TCMID databases were used to screen for the main chemical components and potential targets of the capsule. Integration of OMIM, GeneCards, and DisGeNET databases to obtain BPH-related targets, construction of protein interaction networks using Cytoscape 3.7.2, GO and KEGG pathway enrichment analysis using the Metascape database, validation of the mechanism of action of Huang’e capsule in anti-prostate hyperplasia KEGG screening by immunoblotting.
RESULTS The capsule reduced the wet weight and index of the prostate gland in BPH rats, and significantly improved the morphology of prostate tissue, which resulted in a reduction in the thickness of the epithelium of the prostate, a reduction in the number of papillae protrusions, an increase in the area of the gland lumen, and reduced the expression of prostate-specific antigens in BPH model rats. The 85 active ingredients of the Huang’e capsule were screened; 137 potentially effective targets were related to BPH, and the relevant targets were TP53, AKT1, ESR1, CASP3, JUN, MYC, TNF, and IL6.The active ingredients for the treatment of BPH were quercetin, kaempferol, luteolin and β-sitosterol, which yielded 2193 GO functions and 195 KEGG functions. The key signalling pathways of anti-BPH were mainly involved in the phosphatidylinositol 3-kinase, PI3K, protein kinase B, and AKT signalling pathways. The key anti-BPH signalling pathways are mainly related to the PI3K/AKT signalling pathway. The protein expression in prostate tissues was verified, it was found that Huang’e capsules could significantly reduce the expression of PI3K/AKT/mTOR signalling pathway-related proteins in prostate tissues.
CONCLUSION Huang’e capsule can improve BPH through PI3K/AKT/mTOR signalling pathway.

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