Jun Deng1, Jiajia Qin2, Yi Cai3, Xiaobin Zhong2, Xu Zhang4, Shanshan Yu1. 1. Department of Pharmacy, Zhujiang Hospital, Southern Medical University, Guangdong, Guangzhou, People's Republic of China. 2. Affiliated Langdong Hospital, Guangxi Medical University, Guangxi, Nanning, People's Republic of China. 3. Department of Key Laboratory of Molecular Clinical Pharmacology and Fifth Affiliated Hospital, Guangzhou Medical University, Guangdong, Guangzhou, People's Republic of China. 4. Department of Pediatric Cardiology, Guangdong Cardiovascular Institute and Guangdong General Hospital and Guangdong Academy of Medical Sciences, Guangdong, Guangzhou, People's Republic of China.
Abstract
PURPOSE: The aim of this study is to investigate the potential roles of Rutaecarpine (Rut) in hypoxia-induced human pulmonary artery smooth muscle cells (HPASMCs) model. METHODS: HPASMCs were cultured with or without hypoxia followed by Rut administration. Cytotoxicity and cell proliferation were assessed by CCK-8 and Cell counting method. Flow cytometry was used for the measurement of cell apoptosis rates. The mRNA expression of hypoxia-induced factor (HIF)-1α and protein levels of HIF-1α, p53, p21, erythropoietin, and vascular endothelial growth factor were determined by quantitative real-time polymerase chain reaction and Western blot, respectively. RESULTS: Rut inhibited the proliferation of HPASMCs with IC50 value of 43.5 μmol·L. Hypoxia significantly increased proliferation and decreased apoptosis in HPASMCs, whereas Rut rescued this phenomenon at the appropriate concentration. Meanwhile, Rut effectively decreased the protein and mRNA expressions of HIF-1α. Knockdown of HIF-1α expression by small interfering RNA (siRNA) significantly enhanced the proapoptotic effect rather than antiproliferation effect of Rut in HPASMCs. Moreover, Rut simultaneously reduced proliferating cell nuclear antigen protein expression, whereas increased p53 and p21 protein levels. However, no significant difference was observed in the protein levels of vascular endothelial growth factor and erythropoietin. CONCLUSIONS: Our results demonstrated that Rut exerted protective effects on HPASMCs against hypoxia partly through the HIF-1α-dependent signaling pathway.
PURPOSE: The aim of this study is to investigate the potential roles of Rutaecarpine (Rut) in hypoxia-induced human pulmonary artery smooth muscle cells (HPASMCs) model. METHODS: HPASMCs were cultured with or without hypoxia followed by Rut administration. Cytotoxicity and cell proliferation were assessed by CCK-8 and Cell counting method. Flow cytometry was used for the measurement of cell apoptosis rates. The mRNA expression of hypoxia-induced factor (HIF)-1α and protein levels of HIF-1α, p53, p21, erythropoietin, and vascular endothelial growth factor were determined by quantitative real-time polymerase chain reaction and Western blot, respectively. RESULTS:Rut inhibited the proliferation of HPASMCs with IC50 value of 43.5 μmol·L. Hypoxia significantly increased proliferation and decreased apoptosis in HPASMCs, whereas Rut rescued this phenomenon at the appropriate concentration. Meanwhile, Rut effectively decreased the protein and mRNA expressions of HIF-1α. Knockdown of HIF-1α expression by small interfering RNA (siRNA) significantly enhanced the proapoptotic effect rather than antiproliferation effect of Rut in HPASMCs. Moreover, Rut simultaneously reduced proliferating cell nuclear antigen protein expression, whereas increased p53 and p21 protein levels. However, no significant difference was observed in the protein levels of vascular endothelial growth factor and erythropoietin. CONCLUSIONS: Our results demonstrated that Rut exerted protective effects on HPASMCs against hypoxia partly through the HIF-1α-dependent signaling pathway.