Department of Dermatology, The First Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
Department of Dermatology, Guangzhou Red Cross Hospital, Medical College, Jinan University, Guangzhou, 510220, China.
J Pharmacol Sci. 2019 May;140(1):33-42. doi: 10.1016/j.jphs.2019.04.001. Epub 2019 Apr 15.
The present study aimed to investigate the functional components from Bushen Yijing Formula and their inhibition of endothelial-mesenchymal transition (EndMT) and fibrosis in human umbilical vascular endothelial cells (HUVECs). HUVEC fibrosis was induced by treatment of transforming growth factor β (TGF-β) as the cellular model. Expression of EndMT biomarker gene and cofactors were determined by quantitative real-time-PCR, western blotting, and immunofluorescence. Angiogenesis capacity of vein endothelial cells was evaluated using tube formation assay. Ursolic acid and drug-contained serum ameliorated EndMT biomarker gene expression changes and angiogenesis capacity suppression induced by TGF-β treatment. Slug, Snail, and Twist gene expression and phosphorylation of mammalian target of rapamycin (mTOR) and AKT altered by TGF-β in HUVECs were suppressed by ursolic acid and drug-contained serum. Treatment with the mTOR signaling pathway inhibitor, rapamycin, inhibited the phosphorylation of mTOR and AKT, decreased Snail and Vimentin protein levels, and increased VE-cad protein levels. Overexpression of Snail gene promoted expression of EndMT-related genes and suppressed angiogenesis in HUVECs, which were attenuated by application of ursolic acid and drug-contained serum. Ursolic acid from Bushen Yijing Formula inhibits human umbilical vein endothelial cell EndMT and fibrosis, mediated by AKT/mTOR signaling and Snail gene expression.
本研究旨在探讨补肾益精方的功能成分及其对人脐静脉内皮细胞（HUVEC）内皮-间质转化（EndMT）和纤维化的抑制作用。采用转化生长因子β（TGF-β）处理作为细胞模型诱导 HUVEC 纤维化。通过定量实时 PCR、Western blot 和免疫荧光测定 EndMT 生物标志物基因和辅助因子的表达。通过管形成试验评估静脉内皮细胞的血管生成能力。熊果酸和含药血清改善了 TGF-β 处理诱导的 EndMT 生物标志物基因表达变化和血管生成能力抑制。熊果酸和含药血清抑制了 TGF-β 诱导的 Slug、Snail 和 Twist 基因表达和哺乳动物雷帕霉素靶蛋白（mTOR）和 AKT 的磷酸化。mTOR 信号通路抑制剂 rapamycin 处理抑制了 mTOR 和 AKT 的磷酸化，降低了 Snail 和波形蛋白蛋白水平，增加了 VE-cad 蛋白水平。Snail 基因的过表达促进了 HUVEC 中 EndMT 相关基因的表达，并抑制了血管生成，而熊果酸和含药血清的应用则减弱了这种作用。补肾益精方中的熊果酸通过 AKT/mTOR 信号和 Snail 基因表达抑制人脐静脉内皮细胞 EndMT 和纤维化。
