神经调节素1β对氧糖剥夺/复氧诱导PC12细胞损伤的保护作用及其机制  

PROTECTIVE EFFECT OF NEUREGULIN1βON OXYGEN-GLUCOSE DEPRIVATION/REOXYGENATION-INDUCED PC12 CELL INJURY AND ITS MECHANISM

作　　者：孙珊珊 于曦 翟秋月 于竹芹[3] SUN Shanshan;YU Xi;ZHAI Qiuyue;YU Zhuqin(Department of Neurology,Qilu Hospital of Shandong University(Qingdao),Qingdao 266035,China)

机构地区：[1]山东大学齐鲁医院(青岛)神经内科,山东青岛266035 [2]青岛大学医学部中西医结合中心 [3]青岛大学医学院松山医院内科

出　　处：《精准医学杂志》2023年第6期485-489,493,共6页Journal of Precision Medicine

基　　金：青岛大学医疗集团科研专项(2021-36)。

摘　　要：目的探讨神经调节素1β(NRG1β)对氧糖剥夺/复氧(OGD/R)诱导的PC12细胞损伤的保护作用及其机制。方法建立PC12细胞OGD/R模型,随机分为对照组(常规培养)、模型组(OGD/R处理)以及干预组(给予OGD/R处理+NRG1β干预),采用细胞计数试剂盒(CCK-8)检测各组细胞的活力,采用荧光强度分析技术检测各组细胞活性氧(ROS)水平,采用荧光分光光度法检测各组细胞丙二醛(MDA)水平及还原型谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)吸光度比值,采用透射电镜观察各组细胞线粒体损伤情况,采用Western blot方法检测各组细胞中谷胱甘肽过氧化物酶4(GPX4)的表达情况。结果与对照组比较,模型组的细胞活力显著下降(t=25.76,P<0.01),ROS水平显著升高(t=12.43,P<0.01),GSH/GSSG吸光度比值显著降低(t=9.17,P<0.01),MDA水平显著升高(t=29.46,P<0.01);与模型组比较,干预组的细胞活力显著升高(t=8.03,P<0.01),ROS水平显著降低(t=10.34,P<0.01),GSH/GSSG吸光度比值显著升高(t=15.71,P<0.01),MDA水平显著降低(t=2.96,P<0.05)。透射电镜结果显示,与对照组相比,模型组线粒体损伤增加,NRG1β干预后减缓了线粒体损伤。Western blot方法显示,模型组GPX4表达水平较对照组显著降低(t=23.06,P<0.01),而干预组较模型组显著升高(t=6.07,P<0.05)。结论NRG1β可通过影响铁死亡关键蛋白GPX4的表达,缓解OGD/R诱导的PC12细胞损伤。Objective To investigate the protective effect of neuregulin1β(NRG1β)on PC12 cell injury induced by oxygen-glucose deprivation/reoxygenation(OGD/R)and its mechanism.Methods OGD/R models of PC12 cells were established and randomly divided into control group(routinely cultured),model group(OGD/R-treated),and intervention group(treated with OGD/R and intervened by NRG1β).The cell viability of each group was measured by cell counting kit-8.The levels of reactive oxygen species(ROS)were measured by fluorescence intensity analysis.The levels of malondialdehyde(MDA)and the absorbance ratio of reduced glutathione/oxidized glutathione(GSH/GSSG)were measured by fluorescence spectrophotometry.Mitochondrial damage in each group was observed under a transmission electron microscope.The expression of glutathione peroxidase 4(GPX4)was measured by Western blot.Results Compared with the control group,the model group had significantly decreased cell viability(t=25.76,P<0.01),a significantly increased ROS level(t=12.43,P<0.01),a significantly decreased absorbance ratio of GSH/GSSG(t=9.17,P<0.01),and a significantly increased MDA level(t=29.46,P<0.01).Compared with the model group,the intervention group showed significantly increased cell viability(t=8.03,P<0.01),a significantly decreased ROS level(t=10.34,P<0.01),a significantly increased absorbance ratio of GSH/GSSG(t=15.71,P<0.01),and a significantly decreased MDA level(t=2.96,P<0.05).Transmission electron microscopy results showed that mitochondrial damage was increased in the model group compared with the control group,and NRG1βintervention alleviated mitochondrial damage.Western blot analysis showed that the GPX4 protein expression was significantly lower in the model group than in the control group(t=23.06,P<0.01),and significantly higher in the intervention group than in the model group(t=6.07,P<0.05).Conclusion NRG1βcan alleviate PC12 cell injury induced by OGD/R through regulating the expression of GPX4,a key protein of ferroptosis.

关 键 词：神经调节蛋白1 PC12细胞 铁死亡 谷胱甘肽过氧化酶 氧糖剥夺/复氧 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]