梓葛冻干粉针剂对人脐静脉内皮细胞体外缺血再灌注模型损伤的保护作用

Protective Effects of Zige Lyophilized Powder on Human Umbilical Vein Endotheilial Cells Ischemia-Reperfusion Injury in vitro

目的:研究中药单体组方梓葛冻干粉针剂对人脐静脉内皮细胞(HUVECs)体外缺血再灌注模型损伤的保护作用。方法:体外培养HUVECs,采用氧糖剥夺(Krebs液)方法建立缺氧-复氧模型以模拟体内缺血再灌注损伤。分为正常组、模型组、尼莫地平组、梓葛低、中、高剂量组,复氧时正常组及模型组给予生理盐水,余组给予相应的药物干预后,测定细胞活力、细胞内超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量、一氧化氮(NO)含量、细胞外乳酸脱氢酶(LDH)释放量;分析细胞凋亡相关蛋白Bcl-2、Bax、Caspase-3的表达。结果:与正常组相比,模型组A值、SOD活性显著性降低(P<0.05);MDA和NO含量以及LDH释放量均显著性升高(P<0.05),Caspase-3和Bax蛋白表达显著升高,Bcl-2蛋白表达显著下降,Bcl-2/Bax显著降低(P<0.01)。与模型组比较,梓葛低剂量组A值显著升高(P<0.05);梓葛低、中剂量组MDA和LDH含量显著降低(P<0.05),Bcl-2蛋白表达水平显著升高(P<0.05,P<0.01);梓葛低剂量组NO含量显著降低(P<0.05);梓葛冻干粉针剂的各剂量组SOD活性均显著提高(P<0.05),Bax蛋白水平显著降低(P<0.05,P<0.01),Bcl-2/Bax均显著提高(P<0.05,P<0.01);梓葛中、高剂量组Caspase-3蛋白表达水平显著降低(P<0.01)。结论:梓葛冻干粉针剂可显著拮抗缺氧-复氧对人脐静脉内皮细胞的损伤,其机制与抑制细胞自由基的产生、提高抗氧化能力及调节凋亡相关蛋白表达密切相关。

Objective：To study the protective effects of Zige l yophilized powder on human umbilical vein endotheilial cells （HUVECs）subjected to oxygen-glucose deprivation （OGD）/reperfusion in vitro. Method：HUVECs were exposed to OGD to imitate ischemia-reperfusion in vitro. HUVECs were divided into normal group,model group,nimodipine group,Zige lyophilized powder low, middle and high dosage group.was applied.During reoxgenation, HUVECs in normal group and model group were treated with saline.HUVECs in the other groups were interfered with corresponding drugs.Cell vitality,activity of SOD,contents of MDA, NO and LDH were detected.The expression of apoptotic-related protein Bcl-2, Bax and Caspase-3 were measured. Results： Compared with normal group, cell vitality, activity of SOD, expression of protein Bcl-2, ratio of Bcl-2/ Bax were decreased,contents of MDA, NO, LDH,expressions of Bax and Caspase-3 were increased in model group.Compared with model group,cell vitality was increased, content of NO was decreased in Zige lyophilized powder low dosage group, contents of MDA, LDH were decreased, expression of protein Bcl-2 was increased in Zige lyophilized powder low and middle dosage groups;expression of protein Caspase-3 was decreased in Zige lyophilized powder middle and high dosage groups; activity of SOD,ratio of Bcl-2/Bax were increased and expression of Bax was decreased in Zige lyophilized powder low, middle and high dosage groups. Conclusion ：Zige lyophilized powder has a significant therapeutic effect on the hypoxia- reoxygenation injury of cultured HUVECs,which may be related to inhibiting production of free radicals,elevating the ability of antioxidisty and regulating the expression of apoptotic-related protein.