异钩藤碱减轻帕金森病小鼠多巴胺能神经元损伤的作用机制  

Mechanism of action of isorhynchophylline in attenuating dopaminergic neuronal damage in Parkinson's mice

作　　者：刘崇[1] 刘亚婷 张壮 周国平[2] LIU Chong;LIU Yating;ZHANG Zhuang;ZHOU Guoping(The Second Department of Neurology and Internal Medicine,The First People's Hospital of Nanyang City,Nanyang,He'nan 473000,China;Department of Neurosurgery,Nanyang Central Hospital,Nanyang,He'nan 473000,China)

机构地区：[1]南阳市第一人民医院神经内二科,河南南阳473000 [2]南阳市中心医院神经外科,河南南阳473000

出　　处：《安徽医药》2025年第1期23-28,共6页Anhui Medical and Pharmaceutical Journal

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20191454)。

摘　　要：目的探讨异钩藤碱对帕金森病(PD)小鼠多巴胺能神经元损伤及凋亡的影响及其可能作用机制。方法2020年6月至2022年6月选取40只小鼠建立PD模型,造模成功小鼠按照随机数字表法分为模型组、异钩藤碱低、中和高剂量组,每组各10只,另设对照组10只。异钩藤碱低、中和高剂量组小鼠分别灌胃给药20、40、80 mg/kg的异钩藤碱混悬液(0.9%氯化钠溶液配制),对照组及模型组给予等量0.9%氯化钠溶液。采用爬杆实验和旷场实验评定各组小鼠行为学变化;ELISA法检测小鼠血清中白细胞介素-1β(IL-1β)和肿瘤坏死因子α(TNF-α)水平以及脑组织中多巴胺(DA)水平;TUNEL法检测黑质区神经细胞凋亡;逆转录聚合酶链式反应(RT-PCR)法检测黑质区微RNA-376b-3p(miR-376b-3p)、转录因子SP1(SP1)和亮氨酸重复序列激酶2(LRRK2)的mRNA相对表达量;蛋白质印迹法检测黑质区SP1、LRRK2、α-突触核蛋白(α-syn)和酪氨酸羟化酶(TH)蛋白表达情况。结果与对照组(4.58±0.27)μg/L比较,模型组(1.35±0.13)μg/L小鼠DA水平显著降低(P<0.05)。与模型组比较,异钩藤碱低剂量组(1.86±0.17)μg/L、中剂量组(2.56±0.21)μg/L和高剂量组(3.40±0.29)μg/L小鼠DA水平显著升高(均P<0.05),且以异钩藤碱高剂量组最佳。与模型组比较,异钩藤碱低、中和高剂量组小鼠爬杆时间、IL-1β、TNF-α水平、神经细胞凋亡率、SP1和LRRK2 mRNA和蛋白表达水平、α-syn蛋白水平降低,运动总距离、miR-376b-3p以及TH蛋白水平升高(P<0.05),且以异钩藤碱高剂量组效果最佳(P<0.05)。结论异钩藤碱能够在一定程度上改善PD小鼠运动功能障碍,减轻神经炎症反应,降低神经细胞凋亡率,可能是通过影响miR-376b-3p/SP1/LRRK2信号系统发挥作用。Objective To investigate the effects of isorhynchophylline on dopaminergic neuronal damage and apoptosis in Parkinson´s(PD)mice and its possible mechanism of action.Methods From June 2020 to June 2022,forty micewere selected to establish the PD model,and the successful mice were assigned into model group,the low,medium and high dose of isorhynchophylline groups with 10 mice in each group according to the random number table method,and another 10 mice were selected as control group.The mice in the low,medium andhigh dose groups were given 20,40 and 80 mg/kg of isorhynchophylline suspension(prepared with 0.9%sodium chloride solution)by gavage,while the control and model groups were given equal amounts of 0.9%sodium chloride solution.The behavioral changes of mice in each group were evaluated by pole climbing test and open field test.The levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in serum and dopamine(DA)in brain were detected by ELISA.TUNEL method was used to detect neuronal apoptosis in substantia nigra.The mRNA relative expression levels of miR-376b-3p,transcription factor SP1(SP1)and leucine repeat sequence kinase 2(LRRK2)in the substantium nigra region were detected by reverse transcription quantitative polymerase chain reaction(RT-qPCR).The expressions of SP1,LRRK2,α-synuclein(α-syn)and tyrosine hydroxylase(TH)in substantia nigra were detected by Western blotting.Results Compared with the control group(4.58±0.27)μg/L,the model group(1.35±0.13)μg/L showed a significant decrease in mouse DA levels(P<0.05).Compared with the model group,the low-dose group of isorhynchophylline(1.86±0.17)μg/L,the medium-dose group(2.56±0.21)μg/L,and the high-dose group(3.40±0.29)μg/L exhibited significant increases in mouse DA levels(all P<0.05),with the high-dose group of isorhynchophylline showing the best effect.Compared with the model group,the climbing time,IL-1β,TNF-α,neuronal apoptosis rate,the expression levels of SP1 and LRRK2 mRNA and protein,α-syn protein expression were decrease

关 键 词：钩藤属 帕金森病 异钩藤碱 神经元损伤 转录因子SP1 亮氨酸重复序列激酶2 

分 类 号：R285.5[医药卫生—中药学]