基于"肠-脑轴"角度的氟桂利嗪所致药源性帕金森综合征发病机制研究  

Pathogenesis of flunarizine-induced parkinsonism from gut-brain axis perspective

作　　者：丁楠[1] 潘丽欣 练昌林 徐志锋[1] 王玉凯[1] 张分 赵光华 梁晓珏 赖文杰 曾玮琪 陈静娟[1] 张国华[1] Ding Nan;Pan Lixin;Lian Changlin;Xu Zhifeng;Wang Yukai;Zhang Fen;Zhao Guanghua;Liang Xiaojue;Lai Wenjie;Zeng Weiqi;Chen Jingjuan;Zhang Guohua(Department of Neurology,First People's Hospital of Foshan,Foshan 528000,China;Department of Rehabilitation,First People's Hospital of Foshan,Foshan 528000,China;Department of Laboratory,First People's Hospital of Foshan,Foshan 528000,China;Department of Neurology,Sixth Affiliated Hospital of South China University of Technology,Foshan 528200,China)

机构地区：[1]佛山市第一人民医院神经内科,佛山528000 [2]佛山市第一人民医院康复科,佛山528000 [3]佛山市第一人民医院检验科,佛山528000 [4]华南理工大学附属第六医院神经内科,佛山528200

出　　处：《中华神经医学杂志》2024年第4期333-339,共7页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金青年基金项目(8230053942);中国博士后科学基金第73批面上资助项目(2023M730597);广东省医学科学技术研究基金(A2020407)。

摘　　要：目的从"肠-脑轴"角度探讨氟桂利嗪所致药源性帕金森综合征(DIP)的发病机制。方法30只雄性C57BL/6小鼠按随机数字表法分为空白对照组与氟桂利嗪组,每组15只。空白对照组小鼠给予0.1 mL 50%聚乙二醇400+50%生理盐水混合液灌胃,1次/d、持续2周;氟桂利嗪组小鼠给予浓度6 mg/mL氟桂利嗪+50%聚乙二醇400+50%生理盐水混合液灌胃,每日剂量30 mg/kg,1次/d、持续2周。2组小鼠均于开始给药后第1、3、5、10、14天时记录体质量,于第14天时进行转棒实验以评估运动功能,并留取小鼠粪便行16s RNA测序以观察肠道菌群情况,给予伊文思蓝溶液灌胃检测肠道转运功能,随后处死小鼠,取其脑组织、肠组织并制备成匀浆或冰冻切片,应用Western blotting实验检测黑质中多巴胺能神经元表达情况,应用RT-qPCR法检测黑质中炎性因子表达情况,应用免疫荧光染色检测结肠上皮组织中紧密连接蛋白ZO-1、Claudin-5表达情况。结果与空白对照组比较,开始给药后第1、3、5、7、10、14天时氟桂利嗪组小鼠的体质量比(与给药前体质量的比值)均明显更低。与空白对照组比较,氟桂利嗪组小鼠的转棒停留时间明显缩短,跌落时转棒转速明显降低,差异均有统计学意义(P<0.05)。与空白对照组比较,氟桂利嗪组小鼠黑质中酪氨酸羟化酶蛋白水平有所下降,但差异无统计学意义(P>0.05)。与空白对照组比较,氟桂利嗪组小鼠黑质中促炎因子白细胞介素-6、肿瘤坏死因子-α水平明显升高(1.00±0.00 vs.2.79±0.83;1.00±0.00 vs.3.39±1.37),肠道依文思蓝推进率明显降低(80.67%±4.51%vs.50.67%±6.03%),结肠上皮组织中ZO-1、Claudin-5表达明显降低(27.01±1.41 vs.16.32±2.83;37.00±2.80 vs.24.52±2.12),差异均有统计学意义(P<0.05)。空白对照组与氟桂利嗪组小鼠共同存在576种肠道菌群,空白对照组单独存在744种,氟桂利嗪组单独存在634种。基于加权UniFrac距离的主成分分Objective To explore the pathogenesis of flunarizine-induced parkinsonism from gut-brain axis perspective.Methods Thirty male C57BL/6 mice were randomly divided into control group and flunarizine group(n=15).Mice in the control group were given 0.1 mL 50% polyethylene glycol 400+50% saline by gavage once/d for 2 weeks,while mice in the flunarizine group were given 6 mg/mL flunarizine+50% polyethylene glycol 400+50%saline by gavage at a daily dose of 30 mg/kg for 2 weeks.Body mass was recorded 1,3,5,7,10 and 14 d after drug administration,and motor function was assessed by rotarod test 14 d after drug administration;16s RNA sequencing was performed in the feces to observe the intestinal flora;intestinal transit function was detected by Evans blue by gavage;and then,the mice were sacrificed and homogenate or frozen sections(brain and intestinal tissues)were prepared;dopamine-ergic neuron expression was detected by Western blotting;RT-qPCR was applied to detect the expressions of inflammatory factors in the substantia nigra,and immunofluorescent staining was used to detect the expressions of ZO-1 and Claudin-5 in the intestinal epithelial tissues.Results Compared with the control group,the flunarizine group had lower body mass ratio 1,3,5,7,10 and 14 d after drug administration(ratio to body mass before drug administration).Compared with the control group,the flunarizine group had significantly shortened residence time in rod rotating and lower rotational speed when falling(P<0.05).Compared with the control group,the flunarizine group had decreased tyrosine hydroxylase protein in the substantia nigra without significant difference(P>0.05).Compared with the control group,the flunarizine group had significantly increased interleukin-6 and tumor necrosis factor-αin the substantia nigra(1.00±0.00 vs.2.79±0.83;1.00±0.00 vs.3.39±1.37),significantly lower intestinal Evans blue propulsion rate(80.67%±4.51%vs.50.67%±6.03%),and statistically decreased ZO-1 and Claudin-5 expressions in the colonic epithelial tissues(27.

关 键 词：氟桂利嗪 药源性帕金森综合征 肠-脑轴 

分 类 号：R742.5[医药卫生—神经病学与精神病学]