雷公藤甲素对皮质酮诱导小鼠海马神经元细胞损伤的保护作用  

Protective effect of triptolide on corticosterone-induced hippocampal neuron injury in mice

作　　者：柳丽[1] 季凯琳 徐海玲 曹梦怡 苏丹[2] 刘晶[3] LIU Li;JI Kai-lin;XU Hai-ling;CAO Meng-yi;SU Dan;LIU Jing(School of Pharmacy,Changzhou University,Changzhou 213164,Jiangsu,China;Department of Pharmacy,Changzhou No.2 People's Hospial Afiliated to Nanjing Medical University,Changzhou 213100,Jiangsu,China;Department of Cardiology,General Hospital of Eastern Theater Command,PLA,Nanjing 210002,Jiangsu,China)

机构地区：[1]常州大学药学院,常州213164 [2]南京医科大学附属常州市第二人民医院药理科,常州213100 [3]东部战区总医院(原南京军区南京总医院)心内科,南京210002

出　　处：《医学研究生学报》2021年第11期1138-1143,共6页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81773963);江苏省研究生科研与实践创新计划项目(SJCX21_1293);常州市科技局科技计划项目(CJ20209014)。

摘　　要：目的雷公藤甲素已被证实具有神经保护作用,但其能否减轻应激激素皮质酮(CORT)引起的神经损伤尚无报道,文章旨在探讨TP对CORT诱导小鼠海马神经元(HT-22)细胞损伤的保护作用及其可能机制。方法建立CORT诱导的HT-22细胞损伤模型,并将HT-22细胞分为空白对照组,CORT损伤组和TP干预组,在CORT损伤前加入不同浓度TP孵育半小时。在考察PI3K/AKT抑制剂LY294002(LY)的作用时,增加TP+LY干预组,即HT-22细胞先与LY294002(10μmol/L)共孵育0.5h,再进行相应给药处理。采用MTT法检测细胞活力,生化法检测LDH释放.SOD和GSH-Px活性、MDA水平,Ho-echst染色检测细胞凋亡,Western blot检测AKT、CREB和BDNF的蛋白表达。结果与空白对照组SOD[(41.07±5.21)U/mg蛋白]和GSH-Px[(85.09±4.73)U/mg蛋白]以及MDA[(1.25±0.27)nmol/mg蛋白]相比,CORT损伤组SOD[(20.21±2.53)U/mg蛋白]和GSH-Px[(45.72±6.02)U/mg蛋白]活性水平明显降低(P<0.05),MIDA含量[(3.47±0.39)nmol/mg蛋白]显著升高(P<0.01);与CORT损伤组相比,TP干预能显著升高细胞内SOD[(38.70±4.11)U/mg蛋白]和CSH-Px活性[(88.42±5.40)U/mg蛋白],差异有统计学意义(P<0.05)、降低MDA水平[(1.68±0.12)nmol/mg蛋白],差异有统计学意义(P<0.01)。Hochst33258染色结果显示.对照组HT-22的细胞核被染成均一的淡蓝色,与之相比CORT损伤后细胞核被染成亮蓝色,并伴有细胞核固缩、碎裂、染色质凝结等现象;而TP干预组的细胞核形态及染色情况均得到了显著改善,与对照组相比,CORT组细胞凋亡率显著增加Bel-2/Bax比率显著降低(P<0.01),而TP预处理能明显减少细胞凋亡、提高Bcl-2/Bax比率(P<0.01)。与对照组相比,CORT损伤组AKT、CREB的磷酸化水平及BDNF的表达均明显降低,而TP干预显著促进了pAKT、pCREB和BDNF的表达(P<0.05)。与空白对照组比较,CORT损伤组细胞活力下降而LDH释放量增加(P<0.01);与CORT损伤组比较,TP干预组细胞活力升高(P<0.01),而LDH释放量降低(P<0.01);与Objective Triptolide(TP)has been proven to have neuroprotetive ffeets,but its ability to reduce nerve injury induced by the stress hormone corticosterone(CORT)has not been reported.This study aims to explore the protective ffct of TP on CORT-induced hippocampal neuron(HT-22)injury in mice and its possible mechanism.Methods The CORT-induced HT-22 cell injury model was established,and HT-22 cells were divided into blank control group,CORT injury group and TP intervention group.Before CORT injury,different concentrations of TP were added and ineubated for half an hour.In order to investigate the effect of PI3K/AKT inhibitor LY294002(LY),TP±LY intervention group was added,that is,HT-22 cells were co-incubated with LY294002(10μmo/L)for half an hour,and then corresponding drug administration was performed.Cell viability was detected by MTT assay,LDH release,SOD and GSH-PX activities and MDA levels were detected by biochemical assay,cell apoptosis was dected by Hoechst staining,and protein expressions of AKT,CREB and BDNF were detected by Western blot.Results Compared with the blank con-trol group's SOD[(41.07±5.21)U/mg protein],GSH-Px[(85.09±4.73)U/mg protein]and MDA[(1.25±0.27)nmol/mg protein],the activities of SOD[(20.21±2.53)U/mg protein]and GSH-Px[(45.72±6.02)U/mg protein]in CORT injured group were signifi-cantly decreased(P<0.05),while the content of MDA[(3.47±0.39)nmol/mg protein]was significantly increased(P<0.01).Com-pared with CORT injury group,TP intervention significantly increased the activities of SOD[(38.70±4.11)U/mg protein]and GSH-Px[(88.42±5.40)U/mg protein]in cells.There were stistically signifcant dfferences(P<0.05)and decreased MDA level[(1.68±0.12)nmol/mg protein],the diference was staistically significant(P<0.01).The results of Hoechst33258 staining showed that the nuclei of HT-22 in the control group were stained homogeneous light blue,while those of CORT were stained bright blue,accompanied by nuclear pyknosis.fragmentation,chromatin condensation and other phenomena.Compared with the

关 键 词：雷公藤甲素 皮质酮 氧化应激 凋亡 

分 类 号：R971[医药卫生—药品]