宁夏枸杞总黄酮对H2O2损伤人脐静脉内皮细胞的保护作用

Protective Effect of Total Flavonoids from Lycium barbarum on Injured Human Umbilical Vein Endothelial Cells Induced by Hydrogen Peroxide

摘要:

目的: 研究宁夏枸杞总黄酮对过氧化氢(H₂O₂)损伤的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)的保护作用及可能的机制. 方法: 采用H₂O₂诱导HUVEC氧化应激损伤模型.实验分为正常组、1 mmol·L^-1H₂O₂损伤模型组、枸杞总黄酮保护(100,200,400 mg·L^-1) 预孵育24 h组,以及阳性对照(维生素C 20 mg·L^-1)组,预孵育24 h.采用噻唑蓝(MTT)法检测枸杞总黄酮对1 mmol·L^-1 H₂O₂损伤 4 h细胞活性,测定细胞上清液中丙二醛(MDA),乳酸脱氢酶(LDH),超氧化物歧化酶(SOD),一氧化氮(NO). 结果: 与正常组比较,H₂O₂损伤模型组MDA含量、LDH活性均增高,NO生成量、SOD活性明显下降,其差异均具有显著性(P<0.01).枸杞黄酮保护组MDA含量、LDH活性均下降,NO生成量、SOD活性较H₂O₂损伤模型组明显增高(P<0.01),MDA含量、LDH活性的下降程度,NO生成量、SOD活性的增高程度,与枸杞总黄酮呈剂量依赖性. 结论: 宁夏枸杞总黄酮对H₂O₂所致人血管内皮损伤有保护作用,其机制可能与宁夏枸杞总黄酮抑制损伤细胞的脂质过氧化,以及有效提高机体抗氧化酶的活性、清除自由基,促进一氧化氮(NO)的合成有关.

Abstract:

Objective: To study the protection action and possible mechanism of total flavonoids from Lycium barbarum(FLB) on the injured human umbilical vein endothelial cells(HUVEC)induced by hydrogen peroxide(H₂O₂). Method: The HUVEC oxidative stress damage model was established by using H₂O₂.This study was divided into as follows:normal group, 1 mmol·L^-1 H₂O₂ injury group, FLB protective groups(100,200,400 mg·L^-1)incubation for 24 hours and positive control (vitamin C 20 mg·L^-1)group incubation for 24 h. MTT method was used to detect cell activity that was protected by FLB from damage caused by 1 mmol·L^-1 H₂O₂.The levels of malondialdehyde (MDA)and superoxide dismutase (SOD),the activity of lactate dehydrogenase (LDH), the generation of nitric oxide(NO)in the supernatan were measured. Result: The levels of MDA and the activity of LDH were increased,the generation of No and the activity of SOD decreased obviously in H₂O₂ damage model group,the differences are significant (P<0.01).In FLB protection group, the activity of LDH and the levels of MDA were decreased, the generation of NO and the activity of SOD were increased obviously, the differences were significant(P<0.01). There was a dose dependent manner between the decreased degree of MDA, the activity of LDH, the degree of NO generation,the activity of SOD and FLB. Conclusion: FLB has the ability to protect vascular endothelial from damage caused by H₂O₂, its mechanism of the protecting of total FLB may be related to inhibiting lipid peroxidation damage cells, effectively improving the cell's antioxidant enzyme activity, eliminatimg free radicals, and promoting the production of NO.