丁苯酞对帕金森病伴抑郁小鼠黑质纹状体NLRP3和IBA-1表达的影响

doi:10.3969/j.issn.1003-9198.2022.07.006

摘要/Abstract

摘要： 目的观察丁苯酞对PD伴抑郁小鼠抑郁行为的改善效果及对黑质纹状体区炎症小体核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)及小胶质细胞标志蛋白IBA-1表达的影响。方法 C57成年雄性小鼠40只,随机选取10只为正常对照组,其余小鼠腹腔注射MPTP制备PD小鼠模型,再给予慢性不可预知温和应激构建PD伴抑郁小鼠模型,将其随机分为PD伴抑郁组、美多芭治疗组和美多芭联合丁苯酞治疗组,每组各10只。美多芭治疗组行美多芭(6.25 mg/kg)持续灌胃治疗1周;美多芭联合丁苯酞治疗组行美多芭(6.25 mg/kg)和丁苯酞(120 mg/kg)持续灌胃治疗1周。通过强迫游泳测试、悬尾测试和糖水偏好实验对各组小鼠行为进行评价。应用蛋白免疫印迹法检测各组小鼠黑质纹状体内NLRP3及IBA-1的表达情况。应用免疫荧光法观察各组小鼠黑质纹状体组织中小胶质细胞激活情况及多巴胺神经元细胞损伤情况。结果与正常对照组相比,模型小鼠的强迫游泳静止时间及悬尾静止时间延长、糖水偏好率降低、NLRP3和IBA-1的表达量升高,差异均有统计学意义(P<0.05)。与PD伴抑郁组相比,美多芭治疗组及美多芭联合丁苯酞治疗组的强迫游泳静止时间及悬尾静止时间缩短、糖水偏好率增加、NLRP3和IBA-1的表达量下降,且以美多芭联合丁苯酞治疗组更为明显,差异均有统计学意义(P<0.05)。与正常对照组相比,模型小鼠的异常激活小胶质细胞数量明显增多、胞体变大、轴突减少,多巴胺神经元细胞数量明显减少且细胞体萎缩。与PD伴抑郁组相比,美多芭联合丁苯酞治疗组激活小胶质细胞数量明显减少,多巴胺神经元细胞数量有所增多。结论丁苯酞可能通过抑制小胶质细胞及炎症小体的激活、减轻炎症反应来改善PD伴抑郁小鼠的抑郁症状。

关键词: 帕金森病, 抑郁, 丁苯酞, NLRP3, 小胶质细胞

Abstract: Objective To observe the effects of butylphthalide on depressive behavior and the expression of NOD-like receptor protein 3 (NLRP3) and ionised calcium binding adaptor molecule 1 (IBA-1) in the substantia nigra striatum of mice with Parkinson's disease (PD) and depression. Methods Forty male adult C57 mice were divided into normal control group (n=10) and model group (n=30). The model group was given intraperitoneal injection of MPTP first, and then given chronic unpredictable mild stress to construct the PD combined with depression model. Among the 30 model mice, 10 mice were not treated (PD with depression group), 10 mice were administered with metopar (6.25 mg/kg) for one week (metopar group), and the other 10 mice were administered with metopar (6.25 mg/kg) combined with butylphthalide (120 mg/kg) for one week ( metopar combined with butylphthalide group). The behavior of mice was evaluated by forced swimming test, tail hanging test and sugar water preference test. The expression of NLRP3 and IBA-1 in the substantia nigra striatum was detected by Western Blot. Immunofluorescence was used to observe the activation of microglia and the damage of dopamine neurons in the substantia nigra striatum. Results Compared with normal control group, the forced swimming rest time and tail suspension rest time in PD with depression group were prolonged, the preference rate of sugar water was decreased, and the expression of NLRP3 and IBA-1 was increased, with statistically significant differences (all P<0.05). Compared with PD with depression group, the forced swimming resting time and suspended tail resting time were shortened, the sugar water preference rate was increased, and the expression of NLRP3 and IBA-1 was decreased in metobar group and metobar combined with butylphthalide group, especially in metobar combined with butylphthalide group, with statistical differences (all P<0.05). Compared with normal control group, the number of abnormally activated microglia in PD with depression group was significantly increased, the cell body was enlarged and the axon was decreased; The number of dopamine neurons was decreased, and the cell body was atrophied. Compared with PD with depression group, the number of activated microglia was significantly decreased and the number of dopamine neurons was increased in tmedoba combined with butylphthalide group. Conclusions Butylphthalide can improve the depressive symptoms of PD mice with depression by inhibiting the activation of microglia and inflammatory bodies, and reducing the inflammatory response.

Key words: Parkinson's disease, depression, butyphthalide, NOD-like receptor protein 3, microglia

中图分类号:

R742.5

杨浩辉, 刘斌, 李炳翰, 范少凯, 张艳淑. 丁苯酞对帕金森病伴抑郁小鼠黑质纹状体NLRP3和IBA-1表达的影响[J]. 实用老年医学, 2022, 36(7): 670-674.

YANG Hao-hui, LIU Bin, LI Bing-han, FAN Shao-kai, ZHANG Yan-shu. Effects of butylphthalide on NLRP3 and IBA-1 expression in substantia nigra striatum of mice with Parkinson's disease and depression[J]. Practical Geriatrics, 2022, 36(7): 670-674.

参考文献

[1] BYEON H. Development of a depression in Parkinson's disease prediction model using machine learning [J]. World J Psychiatry, 2020, 10(10):234-244.
[2] 曹飞, 王琴琴. 小胶质细胞介导的神经炎症在帕金森病发生发展中的作用[J]. 济宁医学院学报, 2019, 42(4):289-293, 297.
[3] HE Y, HARA H, NUNEZ G. Mechanism and regulation of NLRP3 inflammasome activation[J]. Trends Biochem Sci, 2016, 41(12):1012-1021.
[4] HUANG S, CHEN Z, FAN B, et al. A selective NLRP3 inflammasome inhibitor attenuates behavioral deficits and neuroinflammation in a mouse model of Parkinson's disease[J]. J Neuroimmunol, 2021, 354:577543.
[5] SAVAGE J C, PICARD K, GONZALEZ-IBANEZ F, et al. A brief history of microglial ultrastructure: distinctive features, phenotypes, and functions discovered over the past 60 years by electron microscopy [J]. Front Immunol, 2018, 25(9):803.
[6] GOETZ C G, PAL G. Initial management of Parkinson's disease[J]. BMJ, 2014, 349:g6258.
[7] 罗日鑫, 袁琼茹, 周泳芬, 等. 丁苯酞治疗帕金森病的现状与展望[J]. 临床荟萃, 2018, 33(11):1002-1004.
[8] 侯新鸽, 叶建, 宋允章, 等. 丁苯酞联合舍曲林对合并抑郁的帕金森病患者的效果[J]. 国际精神病学杂志, 2020, 47(3):540-542, 546.
[9] CHEN Y J, WU T T, LI H, et al. Dl-3-n-butylphthalide exerts dopaminergic neuroprotection through inhibition of neuroinflammation[J]. Front Aging Neurosci, 2019, 11:44.
[10] 林臻, 陈洪志, 赵航, 等. MPTP诱导C57BL/6小鼠帕金森模型的制备和评估[J]. 中国比较医学杂志, 2020, 30(8):57-62, 85.
[11] IZCO M, BLESA J, VERONA G, et al. Glial activation precedes alpha-synuclein pathology in a mouse model of Parkinson's disease[J]. Neurosci Res, 2020, 170:330-340.
[12] CHATTERJEE K, ROY A, BANERJEE R, et al. Inflammasome and α-synuclein in Parkinson's disease: a cross-sectional study[J]. J Neuroimmunol, 2020, 338:577089.
[13] LAHOOTI B, CHHIBBER T, BAGCHI S, et al. Therapeutic role of inflammasome inhibitors in neurodegenerative disorders[J]. Brain Behav Immun, 2021, 91:771-783.
[14] WU A G, ZHOU X G, QIAO G, et al. Targeting microglial autophagic degradation in NLRP3 inflammasome-mediated neurodegenerative diseases[J]. Ageing Res Rev, 2021, 65:101202.
[15] QIN Y, QIU J, WANG P, et al. Impaired autophagy in microglia aggravates dopaminergic neurodegeneration by regulating NLRP3 inflammasome activation in experimental models of Parkinson's disease[J]. Brain Behav Immun, 2021, 91:324-338.
[16] 王军峰, 杨宝. 丁苯酞对脑缺血患者的临床疗效及对血清炎症因子的影响[J]. 中外医学研究, 2021, 19(4):52-54.
[17] LI X, SHI M Q, CHEN C, et al. Phthalide derivative CD21 ameliorates ischemic brain injury in a mouse model of global cerebral ischemia: involvement of inhibition of NLRP3[J]. Int Immunopharmacol, 2020, 86:106-714.