Cissus quadrangularis ethanol extract upregulates superoxide dismutase, glutathione peroxidase and endothelial nitric oxide synthase expression in hydrogen peroxide-injured human ECV304 cells.
BACKGROUND
Cissus quadrangularis has been widely used in traditional medicine for the treatment of hemorrhoid. However, the detailed mechanism of antioxidant defense of C. quadrangularis in endothelial cells under oxidative stress remains unclear.


OBJECTIVE
The present study aims to elucidate the protective role of ethanol extract of C. quadrangularis (CQE) including its constituents, quercetin and resveratrol, on hydrogen peroxide (H(2)O(2))-injured human umbilical vein endothelial ECV304 cells.


METHODS
Viability, genotoxicity and protein expression of ECV304 cells were analyzed by MTT, alkaline comet and Western blot, respectively. Production of intracellular reactive oxygen species (ROS) was determined using dichlorofluorescein fluorescence dye.


RESULTS
After exposing cells to CQE containing quercetin and resveratrol, DNA damage was not observed. CQE including quercetin and resveratrol significantly attenuated ROS in H(2)O(2)-injured ECV304 cells in a concentration-dependent manner. The protein expression of superoxide dismutase (Cu/Zn-SOD, Mn-SOD), glutathione peroxidase (GPx) and endothelial nitric oxide synthase (eNOS) increased in the cells treated with CQE, quercetin or resveratrol prior to H(2)O(2) exposure, as compared with control.


CONCLUSIONS
The results provide a molecular mechanism of C. quadrangularis, which could be partially related to quercetin and resveratrol, in restoring ROS in endothelial cells through the upregulation of Cu/Zn-SOD, Mn-SOD, GPx and eNOS.
Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.
Tarat
T
Sapsrithong
Weeraya
W
Kaewprem
Sarunya
S
Tongumpai
Punnee
P
Nusuetrong
Duangdeun
D
Meksuriyen
D000975:Antioxidants / Q000032:analysis / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D032422:Cissus; D020552:Comet Assay; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D000431:Ethanol / Q000737:chemistry; D005979:Glutathione Peroxidase / Q000096:biosynthesis*; D006484:Hemorrhoids; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide; D008519:Medicine, Traditional; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000096:biosynthesis*; D035261:Plant Components, Aerial; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D011794:Quercetin / Q000032:analysis / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D000077185:Resveratrol; D012997:Solvents / Q000737:chemistry; D013267:Stilbenes / Q000032:analysis / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000096:biosynthesis*
D000975:Antioxidants; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D012997:Solvents; D013267:Stilbenes; D000431:Ethanol; D011794:Quercetin; D006861:Hydrogen Peroxide; D005979:Glutathione Peroxidase; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D013482:Superoxide Dismutase; D000077185:Resveratrol