[Protective effect of Zige lyophilized powder on hypoxia/reoxygenation injury of human umbilical vein endothelial cells].
OBJECTIVE
To study the protective effect of single prescription of traditional Chinese medicine Zige lyophilized powder on hypoxia/reoxygenation injury of human umbilical vein endothelial cells (HUVECs).


METHODS
HUVECs were cultured in vitro and Krebs liquid was adopted to establish the hypoxia/reoxygenation injury model. After intervention of Zige lyophilized powder, MTT colorimetric method was used to determine cell activity, the xanthine oxidase method is adopted to detect the activity of superoxide dismutases (SOD) in cells, the thibabituric acid developing method was adopted to determine the content of malondialdehyde (MDA), the nitrate reductase method was used to detect the content of nitric oxide (NO), the 2,4-dinitrophenylhydrazine developing method was adopted to the content of extracellular lactate dehydrogenase (LDH) , and the Western blot method was used to analyze the expression of cell apoptosis-related proteins Bcl-2, Bax and Caspase-3.


RESULTS
Compared with the model group, 12.5 mg x L(-1) of Zige lyophilized powder can enhanced cell viability (P < 0.05). Besides, 49.0, 24.5, 12.5 mg x L(-1) of Zige lyophilized powder prevents the decline of SOD activity during H/R (P < 0.05). Furthermore, 24.5, 12.5 mg x L(-1) Zige lyophilized powder significantly inhibited the increase of MDA and NO content and improved the expression of Bcl-2 protein (P < 0.05, P < 0.01). And 49.0, 24.5, 12.5 mg x L(-1) of Zige lyophilized powder significantly decreased Bax level and up-regulated Bcl-2/Bax ratio. Caspase activation and apoptosis were monitored in the reoxygenated cells. 49.0, 24.5 mg x L(-1) Zige lyophilized powder can decreased Caspase-3 expression (P < 0.01).


CONCLUSIONS
Zige lyophilized powder can prevent H/R induced injury to HUVECs, which may be related to its effect in inhibiting cell peroxidation injury and enhancing cell antioxidant and antiapoptotic capacities
College of Pharmaceutical Sciences, Southwest University, Chongqing Engineering Research Center for Pharmacodynamics Evaluation, Chongqing 400716, China. ccz@swu.edu.cn
Congcong
C
Zhang
Qiang
Q
Ma
Qiang
Q
Xue
Gang
G
Chen
Xiaoyu
X
Xu
D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D015687:Cell Hypoxia / Q000187:drug effects*; D002478:Cells, Cultured; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008315:Malondialdehyde / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D004365:Drugs, Chinese Herbal; D051028:bcl-2-Associated X Protein; D009569:Nitric Oxide; D008315:Malondialdehyde; D007770:L-Lactate Dehydrogenase; D013482:Superoxide Dismutase; D053148:Caspase 3