绞股蓝总苷对人脑微血管内皮氧糖剥夺的影响  

Effect of gypenosides on human brain microvascular endothelial cells induced by oxygen and glucose deprivation

作　　者：解燕昭 杨梅柳 虞武 董立朋[1] 赵景茹[1] 陈彦霞[4] XIE Yanzhao;YANG Meiliu;YU Wu(Department of Neurology,Hebei General Hospital,Hebei,Shijiazhuang 050000,China;不详)

机构地区：[1]河北省人民医院神经内科,石家庄市050000 [2]衡水学院生命科学学院 [3]衡水学院校医院 [4]河北医科大学第二医院内分泌科

出　　处：《河北医药》2023年第23期3525-3528,3534,共5页Hebei Medical Journal

基　　金：国家自然科学基金青年基金项目(编号:82001229);河北省医学科学研究课题计划项目(编号:20210914,20230293);衡水学院高层次人才科研启动基金项目(编号:2019GC18)。

摘　　要：目的探索绞股蓝总苷对氧糖剥夺诱导的人脑微血管内皮细胞(HBMEC)氧化应激反应的影响。方法体外培养HBMEC,使用不同绞股蓝总苷浓度10 mg/L、50 mg/L、100 mg/L、200 mg/L、400 mg/L干预HBMEC,通过CCK8法筛选绞股蓝总苷最佳剂量。建立氧糖剥夺(OGD)模型,将细胞随机分为Control组、OGD组、OGD+GP(绞股蓝总苷)组。Control组用含10%FBS的DMEM培养基常规培养24 h,OGD组、OGD+GP组进行OGD培养24 h,使用荧光显微镜检测活性氧(ROS)水平,Western Blot检测超氧化物歧化酶(SOD)、醌NAD(P)H脱氢酶1(NQO1)蛋白水平,qRT-PCR检测SOD、NQO1的mRNA水平。结果与Control组比较,5组绞股蓝总苷浓度(10 mg/L、50 mg/L、100 mg/L、200 mg/L、400 mg/L)干预后HBMEC活力无明显差异(P>0.05);与OGD组比较,绞股蓝总苷浓度为10 mg/L、50 mg/L、100 mg/L、200 mg/L、400 mg/L的干预后HBMEC活力差异有统计学意义(P<0.05),而绞股蓝总苷浓度为100 mg/L干预后细胞活力较高,因此绞股蓝总苷最佳剂量为100 mg/L。OGD条件下,绞股蓝总苷降低HBMEC的ROS表达;与OGD组比较,使用绞股蓝总苷干预后,HBMEC的SOD和NQO1蛋白及水平mRNA均升高(P均<0.05)。结论绞股蓝总苷对OGD后的HBMEC有抗氧化应激作用,其机制可能与降低ROS,上调SOD、NQO1水平有关。Objective To explore the effect of gypenosides(Gyp)on oxidative stress of human brain microvascular endothelial cells(HBMECs)induced by oxygen and glucose deprivation(OGD).Methods HBMECs were cultured in vitro with Gyp at the concentrations of 10mg/L,50mg/L,100mg/L,200mg/L and 400mg/L,and the optimal concentration was screened by CCK-8 assay.The in vitro OGD model for 24h was created.Then,HBMECs were respectively treated with blank control(cell culture in DMEM containing 10%fetal bovine serum[FBS]),induced with OGD,and OGD induction plus Gyp treatment.Reactive oxygen species(ROS)were detected by fluorescence microscopy.Protein and mRNA levels of superoxide dismutase(SOD)and NAD(P)H quinone dehydrogenase 1(NQO1)were detected by Western Blot and Quantitative reverse transcription polymerase chain reaction(qRT-PCR),respectively.Results There was no significant difference in the cell viability of HBMECs treated with blank control and Gyp at the concentrations of 10mg/L,50mg/L,100mg/L,200mg/L and 400mg/L(P>0.05).There was a significant difference in the cell viability of HBMECs treated with Gyp at the concentrations of 10mg/L,50mg/L,100mg/L,200mg/L and 400mg/L and OGD-induced HBMECs(P<0.05).HBMECs presented the highest viability under the treatment of 100mg/L Gyp,and thus,the optimal concentration of Gyp was determined.Gyp significantly reduced ROS level in OGD-induced HBMECs(P<0.05).Protein and mRNA levels of SOD and NQO1 were significantly higher in HBMECs treated with Gyp plus OGD compared with those induced with OGD(P<0.05).Conclusion Gyp exerts the anti-oxidation property in OGD-induced HBMECs by lowering ROS level and upregulating SOD and NQO1.

关 键 词：绞股蓝总苷 氧化应激 人脑微血管内皮细胞 活性氧 超氧化物歧化酶 醌NAD(P)H脱氢酶1 

分 类 号：R743.33[医药卫生—神经病学与精神病学]