Pei Yu-Qiang, Zheng Yong-Qiu, Ding Yao-Dong, Xu Qi-Xiang, Cao Di, Wu Ya-Ning, Wang Rui, Yang Jia-Xin, Liang Jing, Ma Qian, Ge Hai-Long
Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing, China.
Drug Research and Development Center, School of Pharmacy, Third-Grade Pharmacology Laboratory of State, Administration of Traditional Chinese Medicine, Anhui Provincial Engineering Research Center for Polysaccharide Drugs, Wannan Medical College, Wuhu, China.
Front Pharmacol. 2021 Jan 11;11:581230. doi: 10.3389/fphar.2020.581230. eCollection 2020.
Triptolide (TP), a naturally derived compound from , has been proven effective in protecting against cardiovascular system, but the molecular mechanisms underlying its protective effects are poorly understood. In the current study, we sought to test the potential protective role of TP in the regulation of vascular calcification in a rat model and explore whether TP attenuates medial vascular calcification by upregulating miRNA-204. Vitamin D3 plus nicotine (VDN) was used to induce a vascular calcification (VC) model of rat aorta. Von Kossa and Hematoxylin-Eosin staining were applied to assess the degree of calcification of rat aortas. Calcium content and alkaline phosphatase activity were measured. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was applied to quantify miRNA-204 expression. The localization of runt-related transcription factor-2 (RUNX2) and bone morphogenetic protein-2 (BMP2) expressions were detected by immunohistochemistry and western blotting. Administration of TP greatly reduced vascular calcification in a dose-dependent manner compared with VC controls. The increase in ALP activity and calcium content was ameliorated by TP. Moreover, protein expression levels of BMP2 and RUNX2 were significantly reduced in calcified aortas. MiRNA-204 expression was increased in the TP-treated groups compared with VC controls and the effects of TP were reversed by the intravenous injection of miRNA-204-interfering lentivirus. However, the miRNA-204-overexpressing lentivirus had no additional effects on ALP activity, calcium content, BMP2 and RUNX2 expressions compared with those from TP group. TP inhibited BMP2 and RUNX2 expression and attenuated vascular calcification via upregulating the level of miRNA-204. TP appears to be a potential new therapeutic option for treating vascular calcification.
雷公藤甲素（TP）是一种从天然提取的化合物，已被证明对心血管系统具有保护作用，但其保护作用的分子机制尚不清楚。在本研究中，我们试图在大鼠模型中测试TP在调节血管钙化中的潜在保护作用，并探讨TP是否通过上调miRNA-204来减轻血管中层钙化。采用维生素D3加尼古丁（VDN）诱导大鼠主动脉血管钙化（VC）模型。应用冯·科萨染色和苏木精-伊红染色评估大鼠主动脉钙化程度。测量钙含量和碱性磷酸酶活性。应用定量逆转录聚合酶链反应（qRT-PCR）定量miRNA-204表达。通过免疫组织化学和蛋白质印迹法检测矮小相关转录因子-2（RUNX2）和骨形态发生蛋白-2（BMP2）表达的定位。与VC对照组相比，TP给药以剂量依赖性方式显著降低了血管钙化。TP改善了碱性磷酸酶活性和钙含量的增加。此外，钙化主动脉中BMP2和RUNX2的蛋白表达水平显著降低。与VC对照组相比，TP处理组中miRNA-204表达增加，静脉注射miRNA-204干扰慢病毒可逆转TP的作用。然而，与TP组相比，miRNA-204过表达慢病毒对碱性磷酸酶活性、钙含量、BMP2和RUNX2表达没有额外影响。TP通过上调miRNA-204水平抑制BMP2和RUNX2表达并减轻血管钙化。TP似乎是治疗血管钙化的一种潜在新疗法。
