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Protective Effects of Hydrogen Sulfide in Hypoxic Human Umbilical Vein Endothelial Cells: A Possible Mitochondria-Dependent Pathway
Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai 201203, China
Institute of Biomedical Sciences, Fudan University, Shanghai 201203, China
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 31 December 2012; in revised form: 29 May 2013 / Accepted: 3 June 2013 / Published: 25 June 2013
Abstract: The aim of the study was to investigate the protective effects of sodium hydrosulﬁde (NaHS), a H2S donor, against hypoxia-induced injury in human umbilical vein endothelial cells (HUVECs) and also to look into the possible mechanisms by which H2S exerts this protective effect. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and scratch wound healing assay were chosen to measure the cell viability and migration-promoting effects. The fluorescent probe, DCFH-DA and 5,5',6,6'-Tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide (JC-1) were applied to detect the reactive oxygen species (ROS) level and mitochondrial membrane potential (ΔΨm). Furthermore, western blots were used to measure the expressions of the apoptosis-related proteins. Under hypoxic conditions, 300 μM and 600 μM of H2S could protect HUVECs against hypoxia-induced injury, as determined by MTT assay. Following the treatment of 60 µM NaHS for 18 h, scratch wound healing assays indicated that the scratch became much narrower than control group. After treatment with 60 µM, 120 µM, and 600 µM NaHS, and hypoxia for 30 min, flow cytometry demonstrated that the ROS concentrations decreased to 95.08% ± 5.52%, 73.14% ± 3.36%, and 73.51% ± 3.05%, respectively, compared with the control group. In addition, the JC-1 assay showed NaHS had a protective effect on mitochondria damage. Additionally, NaHS increased Bcl-2 expression and decreased the expression of Bax, Caspase-3 and Caspase-9 in a dose-dependent way. Our results suggest that H2S can protect endothelial cells and promote migration under hypoxic condition in HUVECs. These effects are partially associated with the preservation of mitochondrial function mediated by regulating the mitochondrial-dependent apoptotic pathway.
Keywords: HUVEC; hydrogen sulfide; hypoxia; protective effects; migration-promoting effects; mitochondria; reactive oxygen species; NaHS
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MDPI and ACS Style
Shen, Y.; Guo, W.; Wang, Z.; Zhang, Y.; Zhong, L.; Zhu, Y. Protective Effects of Hydrogen Sulfide in Hypoxic Human Umbilical Vein Endothelial Cells: A Possible Mitochondria-Dependent Pathway. Int. J. Mol. Sci. 2013, 14, 13093-13108.
Shen Y, Guo W, Wang Z, Zhang Y, Zhong L, Zhu Y. Protective Effects of Hydrogen Sulfide in Hypoxic Human Umbilical Vein Endothelial Cells: A Possible Mitochondria-Dependent Pathway. International Journal of Molecular Sciences. 2013; 14(7):13093-13108.
Shen, Yaqi; Guo, Wei; Wang, Zhijun; Zhang, Yuchen; Zhong, Liangjie; Zhu, Yizhun. 2013. "Protective Effects of Hydrogen Sulfide in Hypoxic Human Umbilical Vein Endothelial Cells: A Possible Mitochondria-Dependent Pathway." Int. J. Mol. Sci. 14, no. 7: 13093-13108.