蒙花苷通过抑制circ_0001359/TGF-β1的抗前列腺纤维化研究*

Linarin Attenuates Prostatic Fibrosis Via Inhibiting Circ_0001359/TGF-β1

关键词:  蒙花苷  Circ_0001359  前列腺  纤维化

Key words:  Linarin  Circ_0001359  Prostate  Fibrosis

摘要:

目的 研究蒙花苷对良性前列腺增生腺体纤维化的改善作用,及抑制环状RNA circ_0001359和促纤维化因子TGF-β1水平的作用机制。方法 建立睾酮诱导的小鼠前列腺增生和人前列腺上皮细胞株增殖模型。通过masson和天狼猩红染色,检测羟脯氨酸含量评价蒙花苷对腺体纤维化的改善作用。通过免疫组织化学检测腺体上皮-间质转化(EMT)标记物的表达,实时荧光定量PCR(qPCR)检测腺体EMT转录因子及上皮细胞EMT标记物的mRNAs水平评估蒙花苷对前列腺EMT的影响。通过qPCR和蛋白印迹检测前列腺circ_0001359和TGF-β1的水平,探讨蒙花苷抑制circ_0001359/TGF-β1的可能作用机制。结果 在动物水平,将模型组和正常对照组的小鼠作比较,前者具有明显的前列腺纤维化和EMT特征,且明显提升了TGF-β1的mRNA水平。将蒙花苷组和模型组的小鼠作比较,前者可使TGF-β1的mRNA的水平显著降低,抑制腺体EMT,改善纤维化特征。在细胞水平,50 mg.L-1^的蒙花苷干预能够显著抑制睾酮诱导的上皮细胞增殖和EMT,降低circ_0001359的水平,并抑制TGF-β1的表达。结论 蒙花苷能够通过抑制circ_0001359/TGF-β1而改善前列腺纤维化。

Abstract:

Objective This study was to investigate the effects of Linarin on prostatic fibrosis and the underlying mechanism via inhibiting circ_0001359/TGF-β1. Methods Both the mice prostate hyperplasia model and human epithelial cell line proliferation model were established by testosterone. The anti-fibrosis effect of Linarin was evaluated by masson and sirius red staining, as well as by detecting hydroxyproline content. The regulating function of Linarin on EMT was evaluated by immunohistochemical analysis for glandular epithelial mesenchymal transition (EMT) markers, as well as by real-time fluorescence quantitative PCR (qPCR) analysis for glandular EMT transcription factors and epithelial EMT markers. The possible mechanism of Linarin on inhibiting circ_0001359/TGF-β1 was explored by detecting the levels of circ_0001359 and TGF-β1 via qPCR and western blotting. Results At the animal level, when compared with the normal control group, the model group mice showed significant prostate fibrosis, EMT characteristics and higher mRNA level of TGF-β1. When compared with the model group, Linarin significant reduced the mRNA level of TGF-β1, inhibited glandular EMT and fibrosis. At the cellular level, 50 mg·L-1 Linarin significantly inhibited testosterone-induced epithelial cell proliferation and EMT, reduced circ_0001359 level and decreased TGF--β1 expression. Conclusion Linarin can attenuate prostatic fibrosis via inhibiting circ_0001359/TGF-β1.