Zhang Congcong, Ma Qiang, Xue Qiang, Chen Gang, Xu Xiaoyu
College of Pharmaceutical Sciences, Southwest University, Chongqing Engineering Research Center for Pharmacodynamics Evaluation, Chongqing 400716, China.
Zhongguo Zhong Yao Za Zhi. 2012 Apr;37(7):997-1001.
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).
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.
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).
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
研究中药紫葛冻干粉单味药对人脐静脉内皮细胞（HUVECs）缺氧/复氧损伤的保护作用。
体外培养HUVECs，采用Krebs液建立缺氧/复氧损伤模型。紫葛冻干粉干预后，用MTT比色法测定细胞活性，采用黄嘌呤氧化酶法检测细胞中超氧化物歧化酶（SOD）活性，采用硫代巴比妥酸显色法测定丙二醛（MDA）含量，采用硝酸还原酶法检测一氧化氮（NO）含量，采用2,4-二硝基苯肼显色法测定细胞外乳酸脱氢酶（LDH）含量，并用蛋白质免疫印迹法分析细胞凋亡相关蛋白Bcl-2、Bax和Caspase-3的表达。
与模型组相比，12.5 mg·L⁻¹紫葛冻干粉可提高细胞活力（P < 0.05）。此外，49.0、24.5、12.5 mg·L⁻¹紫葛冻干粉可防止缺氧/复氧（H/R）过程中SOD活性下降（P < 0.05）。此外,24.5、12.5 mg·L⁻¹紫葛冻干粉显著抑制MDA和NO含量的增加，并改善Bcl-2蛋白的表达（P < 0.05，P < 0.01）。49.0、24.5、12.5 mg·L⁻¹紫葛冻干粉显著降低Bax水平，上调Bcl-2/Bax比值。在复氧细胞中监测Caspase激活和凋亡情况。49.0、24.5 mg·L⁻¹紫葛冻干粉可降低Caspase-3表达（P < 0.01）。
紫葛冻干粉可预防H/R诱导的HUVECs损伤，这可能与其抑制细胞过氧化损伤、增强细胞抗氧化和抗凋亡能力有关。
