Protective effects and plausible mechanisms of antler-velvet polypeptide against hydrogen peroxide induced injury in human umbilical vein endothelial cells.
Antler velvet polypeptide (VAP) is a prominent bioactive component of antler velvet. Whereas uncharacterized crude extracts have typically been used in pharmacological studies, in this study, the velvet polypeptide was isolated and purified by acid water extraction, ethanol precipitation, ammonium sulfate fractionation and precipitation, and chromatography, progressively. Human umbilical vein endothelial cells (HUVECs) were induced with H2O2followed purified polypeptide treatment. Cell viability was evaluated by MTT assay. The apoptosis of cells was detected by fluorescence microscopy and flow cytometry. A cell analyzer was used to measure the mitochondrial membrane potential. The intracellular reactive oxygen species (ROS) levels were determined by flow cytometry. Oxidative stress related biochemical parameters were detected, and the expression of apoptosis-related proteins was examined by Western blot analysis. The results indicated that a 7.0 kDa polypeptide (VAP II) was isolated from antler velvet. VAP II enhanced cell viability, decreased cell apoptosis, reversed depolarization of mitochondrial membrane potential, decreased ROS levels, inhibited oxidative stress, and regulated the downstream signaling apoptotic cascade expression caused by H2O2. The protective effects of VAP II on HUVECs suggests a potential strategy for the treatment of cardiovascular disease.
a Department of Biochemistry, Jilin Medical University, Jilin, Jilin 132013, P.R. China.
Wenhe
W
Zhu
a Department of Biochemistry, Jilin Medical University, Jilin, Jilin 132013, P.R. China.
Huiyan
H
Wang
a Department of Biochemistry, Jilin Medical University, Jilin, Jilin 132013, P.R. China.
Wei
W
Zhang
a Department of Biochemistry, Jilin Medical University, Jilin, Jilin 132013, P.R. China.
Na
N
Xu
a Department of Biochemistry, Jilin Medical University, Jilin, Jilin 132013, P.R. China.
Junjie
J
Xu
a Department of Biochemistry, Jilin Medical University, Jilin, Jilin 132013, P.R. China.
Yan
Y
Li
b Institute of Military Veterinary Sciences, Academy of Military Medical Sciences, Changchun, Jilin 130117, P.R. China.
Wensen
W
Liu
a Department of Biochemistry, Jilin Medical University, Jilin, Jilin 132013, P.R. China.
Shijie
S
Lv
D000818:Animals; D000999:Antlers / Q000737:chemistry*; D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D019610:Cytoprotection / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D010455:Peptides / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism
D010455:Peptides; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide
antioxidant; antioxydant; antler velvet polypeptides; apoptose; apoptosis; oxidative stress; polypeptide de bois de velours; stress oxydatif