CB2受体激活对慢性PD模型小鼠运动功能和黑质胶质细胞活化影响
Effect of cannabinoid type 2 receptor activation on motor function and glial cell activation in the substantia nigra in a mouse model of chronic Parkinson's disease
刘欣宇 1张丽 1马泽刚1
作者信息
- 1. 青岛大学基础医学院生理学教研室,山东青岛 266071
- 折叠
摘要
目的 通过行为学、免疫印迹技术及免疫组织化学技术探讨大麻素Ⅱ型(CB2)受体对1-甲基-4-苯基吡啶(MPTP)诱导的慢性帕金森病(PD)模型小鼠运动功能、黑质(SN)区酪氨酸羟化酶(TH)蛋白表达及小胶质细胞和星形胶质细胞活化的影响.方法 将30只8周龄雄性C57BL/6野生型(WT)小鼠随机分为WT对照组(A组)、WT MPTP 组(B 组)、WT CB2 受体激动剂(JWH133)组(C 组)、WT MPTP+JWH133 组(D 组)和 WT MPTP+JWH133+CB2受体拮抗剂(AM630)组(E组),12只8周龄雄性CB2受体敲除(CB2-KO)C57BL/6小鼠随机分为CB2-KO对照组(F组)和CB 2-KO MPTP组(G组).模型组小鼠首先腹腔注射20 μg/(kg·d)AM630和(或)10 μg/(kg·d)JWH133,每天1次,连续注射30 d;然后腹腔注射30 mg/(kg·d)的MPTP,每周2次,持续4周.对照组小鼠腹腔注射等量的生理盐水.应用行为学实验检测各组小鼠爬杆与转棒时间,免疫印迹技术检测SN区TH蛋白的表达,免疫组织化学染色检测SN区小胶质细胞和星形胶质细胞数量和形态变化.结果 与A组相比,B组小鼠爬杆时间增加,转棒时间减少;与B组相比,D组小鼠爬杆时间减少,转棒时间增加;与D组相比,E组小鼠爬杆时间增加,转棒时间减少;与F组相比,G组小鼠爬杆时间增加,转棒时间减少.上述差异具有统计学意义(F=29.70、45.45,q=4.87～18.09,P＜0.05).与A组相比,B组小鼠SN区TH蛋白表达水平下降;与B组相比,D组小鼠SN区TH蛋白表达水平上调;与D组相比,E组小鼠SN区TH蛋白表达水平下降;与F组相比,G组小鼠SN区TH蛋白表达水平下降.上述差异具有统计学意义(F=24.88,q=5.09～8.88,P＜0.001).小鼠SN区活化的小胶质细胞和星形胶质细胞计数显示,与A组相比,B组明显增加;与B组相比,D组明显减少;与D组相比,E组明显增加;与F组相比,G组明显增加.上述差异具有统计学意义(F=269.80、708.50,q=13.29～54.78,P＜0.01).结论 激活CB2受体能够改善MPTP诱导的慢性PD模型小鼠的运动功能障碍,抑制小鼠SN区小胶质细胞和星形胶质细胞的活化.
Abstract
Objective To investigate the effect of cannabinoid type 2(CB2)receptor on motor function,the protein expres-sion of tyrosine hydroxylase(TH),and the activation of microglial cells and astrocytes in the substantia nigra(SN)in a mouse model of chronic Parkinson's disease(PD)induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)based on beha-vioristics,Western blotting,and immunohistochemistry.Methods A total of 30 wild-type(WT)male C57BL/6 mice,aged 8 weeks,were randomly divided into WT control group(group A),WT MPTP group(group B),WT CB2 receptor agonist JWH133 group(group C),WT MPTP+JWH133 group(group D),and WT MPTP+JWH133+CB2 receptor antagonist AM630 group(group E),and 12 CB2 receptor-knockout(CB2-KO)male C57BL/6 mice were randomly divided into CB2-KO control group(group F)and CB2-KO MPTP group(group G).The mice in the model group were given intraperitoneal injection of 20 μg/(kg·d)AM630 and/or 10 μg/(kg·d)JWH133 once a day for 30 consecutive days,followed by intraperitoneal injection of 30 mg/(kg d)MPTP twice a week for four weeks,while those in the control group were given intraperitoneal injection ofan equal volume of normal saline.Behavioral experiments were used to measure the time of poleclimbing and the time spent on the rotating rod;Western blotting was used to measure the protein expression of TH in the SN;immunohistochemical staining was used to observe changes in the number and morphology of microglial cells and astrocytes in the SN.Results Compared with group A,group B had a significant increase in the time of poleclimbing and a significant reduc-tion in the time spent on the rotating rod;compared with group B,group D had a significant reduction in the time of poleclimbing and a significant increase in the time spent on the rotating rod;compared with group D,group E had a significant increase in the time of poleclimbing and a significant reduction in the time spent on the rotating rod;compared with group F,group D had a signi-ficant increase in the time of poleclimbing and a significant reduction in the time spent on the rotating rod(F=29.70,45.45;q=4.87-18.09;P＜0.05).Compared with group A,group B had a significant reduction in the protein expression level of TH in the SN of mice;compared with group B,group D had a significant increase in the protein expression level of TH;compared with group D,group E had a significant reduction in the protein expression level of TH;compared with group F,group G had a significant re-duction in the protein expression level of TH(F=24.88,q=5.09-8.88,P＜0.001).Compared with group A,group B had signifi-cant increases in the numbers of activated microglial cells and astrocytes in the SN of mice;compared with B,group D had signifi-cant reductions in these numbers;compared with group D,group E had significant increases in these numbers;compared with group F,group G had significant increases in these numbers(F=269.80,708.50;q=13.29-54.78;P＜0.01).Conclusion Ac-tivation of CB2 receptor can improve dyskinesia and inhibit the activation of microglial cells and astrocytes in the SN of mice with MPTP-induced chronic PD.
关键词
受体,大麻酚,CB2/帕金森病/1-甲基-4-苯基吡啶/旋转棒性能试验/黑质/小神经胶质细胞/星形细胞/小鼠,近交C57BLKey words
receptor,cannabinoid,CB2/Parkinson disease/1-methyl-4-phenylpyridinium/rotarod performance test/sub-stantia nigra/microglia/astrocytes/mice,inbred C57BL引用本文复制引用
基金项目
山东省重点研发计划资助项目(2019GSF10-8095)
出版年
2024
国家科技期刊平台
NSTL
万方数据