圣草次苷调控MPTP诱导的慢性小鼠帕金森疾病和机制研究  

Study on eriocitrin alleviating MPTP-induced chronic Parkinson′s disease in rats and its mechanism

作　　者：张晓愉[1] 张昆鹏[2] 王建民[1] 赵晓辉[1] ZHANG Xiao-Yu;ZHANG Kun-Peng;WANG Jian-Min;ZHAO Xiao-Hui(Department of Neurology,Xingtai People′s Hospital,Xingtai 054000,China)

机构地区：[1]邢台市人民医院神经内科,邢台054000 [2]邢台市人民医院肝胆外科,邢台054000

出　　处：《中国免疫学杂志》2020年第3期369-374,共6页Chinese Journal of Immunology

基　　金：河北省重点研发计划自筹项目(172777205)

摘　　要：目的:探究圣草次苷减轻1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的慢性小鼠帕金森疾病和机制。方法:选取75只小鼠采用随机分组法分为:健康对照组、模型组、圣草次苷低剂量组(8 mg/kg)、圣草次苷中剂量组(16 mg/kg)、圣草次苷高剂量组(32 mg/kg),每组各15只;腹腔注射20 mg/(kg·d)的MPTP溶液,持续一周制成帕金森小鼠模型;给药组小鼠分别使用对应剂量的圣草次苷干预2周。使用旷场实验、悬挂实验、游泳实验检测小鼠行为;使用HE染色和TUNEL染色检测小鼠神经元细胞的凋亡情况;使用蛋白质印迹法检测小鼠脑组织Bax、Bcl-2和Caspase-3、Caspase-9的表达情况;使用试剂盒检测小鼠黑质中SOD、MDA、LDH、GSH的含量;使用ELISA检测小鼠外周血中IL-6、TNF-α、iNOS和IL-10的含量;使用蛋白质印迹检测信号通路Nrf2、HO-1、NQO1的表达情况。结果:HE染色和TUNEL染色结果显示相比空白对照组,模型组小鼠神经细胞凋亡率显著升高(P<0.05),相比模型组,使用圣草次苷处理各组小鼠神经细胞凋亡率显著降低(P<0.05);相比空白对照组,模型组小鼠旷场移动路程、旷场中心区域活动时间、游泳时间和悬挂时间、SOD水平、Bcl-2、Nrf2、HO-1、NQO1蛋白表达水平显著降低(P<0.05),Bax、Caspase-3和Caspase-9蛋白表达水平、MDA、LDH、GSH、TNF-α、IL-6、iNOS水平显著升高(P<0.05),IL-10无显著变化(P>0.05);相比模型组圣草次苷低剂量组Nrf2、HO-1、NQO1蛋白表达水平显著升高,Bax/Bcl-2显著降低(P<0.05),其余各指标均无显著变化(P>0.05);相比模型组,圣草次苷中剂量组和圣草次苷高剂量组旷场移动路程、旷场中心区域活动时间、游泳时间、SOD、IL-10、Nrf2、HO-1、NQO1蛋白表达水平显著升高(P<0.05),Bax/Bcl-2、Caspase-3和Caspase-9蛋白表达水平、MDA、LDH、GSH、TNF-α、IL-6、iNOS显降低(P<0.05)。结论:圣草次苷可以通过抑制氧化应激、炎症反应,神经细胞的凋亡并激Objective:To explore eriocitrin alleviating chronic Parkinson′s disease(PD)in rats induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)and its mechanism.Methods:The random group method was applied to divided 75 mice into healthy control group,model group,low-dose eriocitrin(8 mg/kg)group,middle-dose eriocitrin(16 mg/kg)group and high-dose eriocitrin(32 mg/kg)group,15 cases in each group.They were given intraperitoneal injection of 20 mg/(kg·d)MPTP solution for one week to make PD mouse models.The mice in administration group were given corresponding doses of eriocitrin for 2 weeks.Their behaviors were tested by field test,suspension test and swimming test.The apoptosis of neuronal cells was detected by HE staining and TUNEL staining.The expression of Bax,Bcl-2,Caspase-3 and Caspase-9 in brain tissue was detected by Western blot.The kit was applied to detect contents of SOD,MDA,LDH and GSH in substantia nigra of mice.ELISA was applied to detect contents of IL-6,TNF-α,iNOS and IL-10 in peripheral blood.The expression of signal pathways Nrf2,HO-1 and NQO1 was detected by Western blot.Results:The results of HE staining and TUNEL staining showed that compared with blank control group,apoptosis rate of neuronal cells was significantly increased in model group(P<0.05).Compared with model group,apoptosis rate of neuronal cells was significantly decreased in all groups treated with eriocitrin(P<0.05).Compared with blank control group,movement distance in field,activity time in field center area,swimming time and suspension time,SOD level,expression level of Bcl-2,Nrf2,HO-1 and NQO1 protein were significantly decreased in model group(P<0.05),while expression levels of Bax,Caspase-3 and Caspase-9,levels of MDA,LDH,GSH,TNF-α,IL-6 and iNOS were significantly increased(P<0.05).There was no significant change in IL-10(P>0.05).Compared with model group,expression levels of Nrf2,HO-1 and NQO1 protein were significantly increased in low-dose eriocitrin group,while Bax/Bcl-2 was significantly decreased(P<0.05).There w

关 键 词：帕金森 圣草次苷 凋亡 氧化应激 炎症反应 

分 类 号：R35[医药卫生—基础医学]