蒙花苷通过抑制Hsa_circ_0001359/TGF-β1抗前列腺纤维化的研究

Linarin Anti-prostatic Fibrosis via Inhibiting Hsa_circ_0001359/TGF-β1

目的观察蒙花苷对Hsa_circ_0001359水平和TGF-β1表达的抑制效应,揭示其改善良性前列腺增生腺体纤维化的作用机制。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,Masson和天狼猩红染色观察腺体胶原沉积,分光光度法检测腺体羟脯氨酸含量,免疫组织化学检测腺体上皮-间质转化(epithelialmesenchymal transition,EMT)标记物的表达,实时荧光定量PCR检测腺体EMT转录因子及上皮细胞EMT标记物mRNAs,以及Hsa_circ_0001359的水平,蛋白印迹法检测TGF-β1的表达。结果在动物水平,与正常对照组比较,模型组前列腺胶原沉积和EMT特征明显,羟脯氨酸的含量以及TGF-β1的mRNA水平显著升高(P<0.01);与模型组比较,各蒙花苷组均改善腺体的胶原沉积与EMT,显著降低羟脯氨酸的含量以及TGF-β1的mRNA水平(P<0.01)。在细胞水平,50 mg·L^(−1)的蒙花苷显著抑制睾酮诱导的上皮细胞增殖和EMT,降低Hsa_circ_0001359的水平,抑制TGF-β1的表达(P<0.01或<0.05)。结论蒙花苷能够通过抑制Hsa_circ_0001359/TGF-β1而改善前列腺纤维化。

OBJECTIVE To investigate the underlying mechanism of linarin on improving benign prostatic hyperplasia glandular fibrosis via observing its effects on inhibiting Hsa_circ_0001359 level and TGF-β1 expression.METHODS Both the mice prostate hyperplasia model and human epithelial cell line proliferation model were established by testosterone.The glandular collagen deposition was evaluated by Masson and sirius red staining.The hydroxyproline content was detected using spectrophotometry.The expression of epithelial mesenchymal transition(EMT)markers were evaluated by immunohistochemical analysis.The levels of glandular EMT transcription factors and epithelial EMT markers mRNAs,as well as the Hsa_circ_0001359 were evaluated by real-time fluorescence quantitative PCR analysis.The expression of TGF-β1 was explored by Western blotting.RESULTS At the animal level,compared with the normal control group,the model group mice showed prostate fibrosis,EMT characteristics,as well as significant higher content of hydroxyproline and mRNA level of TGF-β1(P<0.01).Compared with the model group,linarin inhibited glandular EMT and fibrosis,as well as significant reduced the content of hydroxyproline and mRNA level of TGF-β1(P<0.01).At the cellular level,50 mg·L^(−1) linarin significantly inhibited testosterone-induced epithelial cell proliferation and EMT,reduced Hsa_circ_0001359 level and decreased TGF-β1 expression(P<0.01 or P<0.05).CONCLUSION Linarin can attenuate prostatic fibrosis via inhibiting Hsa_circ_0001359/TGF-β1.