白杨素减轻人脐静脉内皮细胞的氧化应激损伤

Chrysin attenuates oxidative stress in human umbilical vein endothelial cells

目的:观察白杨素对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)氧化应激损伤的保护作用。方法:常规培养HUVEC细胞,分为5组:对照组,不加任何干预;H2O2组,用400μmol/L过氧化氢处理2 h;白杨素高、中、低剂量组,提前用100、50、25μmol/L的白杨素孵育12 h,然后用400μmol/L过氧化氢处理2 h。处理结束后,用MTT法检测活细胞数目,并检测乳酸脱氢酶(LDH)的漏出。收集细胞上清液及细胞裂解液,检测NO、丙二醛(MDA)产量及内皮型一氧化氮合酶(eNOS)、超氧化物歧化酶(SOD)活性。用荧光染料DCEF-DA染色后,酶标仪检测活性氧(ROS)的产量。用荧光定量RTPCR的方法检测黏附分子的表达。结果:过氧化氢导致内皮细胞活细胞数目减少,LDH漏出增多,MDA、ROS产生增多,SOD活性下降,黏附分子表达增多,内皮细胞NO产生减少,eNOS活性下降。提前用白杨素预孵,可以增加细胞存活率,减轻氧化应激损伤,减少黏附分子的表达,eNOS蛋白活性增加,细胞NO的产量增加。结论:白杨素可以减轻HUVEC的氧化应激损伤,减少黏附分子的表达,增加细胞NO的合成能力。

AIM： To observe the protective effects of chrysin against H202-induced oxidative stress injury in human umbilical vein endothelial cells （HUVECs） and the possible mechanism. METHODS： HUVECs were subjected to H202 in the absence or presence of chrysin （25, 50, 100 μmol/L） for 2 h. The 3-（4,5-Dimethyhhiazol-2-yl 1） -2,5 -diphenyltetrazolium bromide （MTT） assay was used to assess cell viability. Reactive oxygen species was detected by dichlorofluorescein diaeetate （DCFH-DA）. The release of lactate dehydrogenase （ LDH ）, the level of malondialdehyde （ MDA ）, NO, superoxide dismutase （SOD） and endothelial nitric oxide synthase （eNOS） in culture media or in cells were assessed using commercial kit according to the manufacturer＇s instructions. The expressions of adhesion molecule mRNA were detected by reversetranscription-quantitative polymerase chain reaction. RESULTS：The results showed that H202 increased the release of LDH, the production of MDA and ROS, the mRNA level of adhesion molecules, decreased the cell viability and the activity of SOD and eNOS, reduced the production of NO. Pretreating HUVECs with chrysin improved cell viability and inhibited the release of LDH, reduced the production of ROS and MDA, alleviated the mRNA level of ad- hesion molecules, elevated the production of NO, improved the activity of SOD and eNOS. CONCLUSION： Chrysin might exert protective effects against oxidative stress in HUVEC.