黄酮哌酯通过抑制TGF-β1减轻双酚A诱导的前列腺纤维化

Flavoxate attenuates bisphenol A-induced prostatic fibrosis by inhibiting TGF-β1

摘要

目的 研究黄酮哌酯对双酚A（BPA）诱导的前列腺纤维化的抑制作用和机制。方法 在动物水平通过每日灌胃10 µg·kg^-1的BPA建立大鼠BPA暴露模型,持续6周后,收集大鼠前列腺组织进行Masson和天狼猩红染色观察各组动物前列腺组织胶原沉积情况,并通过免疫组化和qPCR检测E-钙黏蛋白（E-cad）、波形蛋白（VIM）和α-平滑肌肌动蛋白（α-SMA）评估各组动物前列腺组织上皮–间质转化（EMT）程度。在细胞水平通过用100 nmol·L^-1的BPA孵育人前列腺上皮RWPE-1细胞建立前列腺细胞BPA暴露模型,通过qPCR检测α-SMA、E-cad及胶原相关基因Col1A1和Col3A1的mRNA水平,评价各组细胞胶原分泌和EMT的影响;并通过Western blot检测TGF-β1的蛋白表达水平,探讨黄酮哌酯的作用机制。结果 在动物水平,黄酮哌酯在80或40 mg·kg^-1给药剂量下抑制BPA诱导的大鼠前列腺胶原沉积和EMT。在细胞水平,相较于模型组,50 µg·L^-1的黄酮哌酯显著降低RWPE-1细胞α-SMA、VIM、Col1A1和Col3A1的mRNA水平,并增加E-cad的mRNA水平,抑制TGF-β1的表达。且该效应在一定程度上被TGF-β通路激动剂抵消。结论 黄酮哌酯通过降低TGF-β1的水平,抑制胶原沉积与EMT,改善BPA诱导的前列腺纤维化。

Abstract

Objective To study the inhibitory effect and mechanism of flavoxate on prostatic fibrosis induced by bisphenol A (BPA). Methods At the animal level, the BPA exposure model was established by orally administering 10 µg·kg^-1 BPA daily to rats for 6 weeks. Then, the rat prostates were collected to evaluate tissue collagen deposition by Sirius red and Masson staining, as well as to assess epithelial-mesenchymal transition (EMT) by immunohistochemistry analysis and quantitative real-time PCR (qPCR) analysis for E-cadherin (E-cad), vimentin (VIM) and α-smooth muscle actin (α-SMA). At the cellular level, the BPA exposure model was established by incubating human prostatic epithelial RWPE-1 cells with 100 nmol·L^-1 BPA. The collagen secretion and EMT of RWPE-1 cells were evaluated by qPCR analysis for the mRNA levels of α-SMA, E-cad, Col1A1 and Col3A1. The mechanism of flavoxate was investigated by Western blot analysis for TGF-β1. Results At the animal level, daily treatment with 80 or 40 mg·kg^-1 flavoxate attenuated BPA-induced rat prostate collagen deposition and EMT. At the cellular level, compared with the model group, 50 µg·L^-1 flavoxate significantly downregulated α-SMA, VIM, Col1A1 and Col3A1 while upregulated E-cad in RWPE-1 cells, accompanied by suppression of TGF-β1. These effects were partially reversed by a TGF-β pathway agonist. Conclusion Flavoxate can improve BPA-induced prostatic fibrosis by suppressing TGF-β1 and inhibiting collagen deposition and EMT.

关键词

黄酮哌酯 / 双酚A / 前列腺 / 纤维化 / 上皮-间质转化

Key words

flavoxate / bisphenol A / prostate / fibrosis / epithelial-mesenchymal transition

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基金资助

江汉大学学生科研项目（2024zd061）; 江汉大学校级科研项目资助计划（2023KJZX23）