Nrf2/HO-1信号通路在6-羟基多巴胺帕金森病模型大鼠神经损伤中的作用  

Role of Nrf2/HO-1 signaling pathway in 6-hydroxydopamine-induced nerve injury of Parkinson’s disease model rats

作　　者：王昆[1] 罗小娜 李时光[1] 冯丽君[1] 刘新生[1] WANG Kun;LUO Xiao-na;LI Shi-guang;FENG Li-jun;LIU Xin-sheng(Department of Neurology,Zhengzhou First Peoples' Hospital,Henan 450004,China)

机构地区：[1]郑州市第一人民医院神经内科

出　　处：《中国医药生物技术》2019年第3期237-244,共8页Chinese Medicinal Biotechnology

摘　　要：目的探究核因子E2-相关因子2/血红素加氧酶-1(Nrf2/HO-1)信号通路在6-羟基多巴胺诱导帕金森病(PD)模型大鼠神经损伤中的作用。方法将大鼠分为对照组、PD组、Nrf2激活剂(Hesperin)组、Nrf2抑制剂(Brusatol)组,每组12只,APO旋转诱导实验评估大鼠神经行为学变化并观察各组大鼠异常不自主(AIM)评分;免疫组化检测脑组织黑质中酪氨酸羟化酶(TH)阳性表达情况;透射电镜观察大鼠神经元细胞形态变化;对比各组脑黑组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活性。体外培养SH-SY5Y细胞诱导PD模型细胞,将细胞分为对照组、PD组、Hesperin组、Brusatol组,MTT检测各组细胞活性;流式细胞术检测细胞凋亡情况;免疫印迹法检测脑黑质部以及细胞中Nrf2、HO-1、Bcl2、caspase3、Bax蛋白表达情况。结果与对照组相比,PD组APO诱导圈数、AIM评分均升高,转棒滞留时间缩短,脑组织黑质区TH神经元比例降低,细胞核皱缩,核内异染色质变多呈小块状分布;SOD、GSH-Px、CAT水平降低,MDA水平升高,PD组细胞增殖抑制率、凋亡率升高,脑黑质组织以及细胞中Nrf2、HO-1、Bcl2蛋白表达降低,caspase3、Bax蛋白表达升高,差异均具有显著性(P<0.05);与PD组相比,Hesperin组和PD组APO诱导圈数、AIM评分均降低,转棒滞留时间增长,脑组织黑质区TH神经元比例升高,细胞核形态明显恢复;SOD、GSH-Px、CAT水平升高,MDA水平降低,细胞增殖抑制率、凋亡率降低,脑黑质组织以及细胞中Nrf2、HO-1、Bcl2蛋白表达升高,caspase3、Bax蛋白表达降低,差异均具有显著性(P<0.05);与PD组相比,Brusatol组APO诱导圈数、AIM评分均升高,转棒滞留时间缩短,脑组织黑质区TH神经元比例降低,细胞核严重皱缩;SOD、GSH-Px、CAT水平降低,MDA水平升高,PD组细胞增殖抑制率、凋亡率升高,脑黑质组织以及细胞中Nrf2、HO-1、Bcl2蛋白表达降低,caspase3、BObjective To investigate the role of nuclear factor E2-related factor 2/heme oxygenase-1(Nrf2/HO-1)signaling pathway in 6-hydroxydopamine-induced nerve injury of Parkinson's disease model rats.Methods Rats were divided into control group,PD group,Nrf2 activator group(Hesperin)and Nrf2 inhibitor group(Brusatol),with 12 rats in each group.The neurobehavioral changes of the rats were evaluated by APO rotation induction experiment and the abnormal involuntary(AIM)score of each group of rats was observed.Immunohistochemistry was used to detect the positive expression of tyrosine hydroxylase(TH)in the substantia nigra of brain tissues,and the morphological changes of neurons in rats were observed by transmission electron microscope.The activities of superoxide dismutase(SOD),malondialdehyde(MDA),glutathione peroxidase(GSH-Px)and catalase(CAT)in the substantia nigra tissues of each group were compared.SH-SY5Y cells were cultured in vitro,and the cells were divided into control group,PD group,Hesperin group and Brusatol group.MTT was used to detect cell viability in each group,and apoptosis was detected by flow cytometry.The expression of Nrf2,HO-1,Bcl2,caspase-3,and Bax in the substantia nigra and the cells were detected by Western blot.Results Compared with the control group,the number of APO induction cycles and AIM score were increased and the retention time of rotatory rods was shortened in the PD group.Moreover,the proportion of TH neurons in the substantia nigra of PD model was decreased,and the nuclei were observed to shrink,with more heterochromatin in the nucleus being distributed in small size.Levels of SOD,GSH-Px,and CAT were decreased,while the level of MDA was increased in the model group.The proliferation inhibition rate and apoptosis rate of SH-SY5Y cells were also increased in the PD group.Expressions of Nrf2,HO-1,and Bcl2 in brain substantia nigra and the cells were decreased,while the expression of caspase3 and Bax were increased in the PD model with significant differences(P<0.05 versus control).Compa

关 键 词：羟多巴胺 帕金森病 血红素加氧酶-1 核因子E2-相关因子2 神经损伤 

分 类 号：R742.5[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]