咪唑克生减缓大鼠实验性自身免疫性脑脊髓炎脑损伤的保护作用时间窗研究

doi:10.12092/j.issn.1009-2501.2018.01.001

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (1): 1-7.doi: 10.12092/j.issn.1009-2501.2018.01.001

• 基础研究 • 下一篇

咪唑克生减缓大鼠实验性自身免疫性脑脊髓炎脑损伤的保护作用时间窗研究

厉芳¹，郑浏璞²，翁建龙¹，郭涛¹，宋兴伟¹，郑荣远³

¹杭州市萧山区第一人民医院神经内科，杭州 311200，浙江； ²温州医科大学附属第一医院麻醉科，温州 325000，浙江； ³温州医科大学附属第一医院神经内科，温州 325000，浙江

收稿日期:2017-05-22 修回日期:2017-06-15 出版日期:2018-01-26 发布日期:2018-02-07
通讯作者: 郑荣远，男，本科，教授，博士生导师，研究方向：神经病学，神经药理学。 E-mail:zhengry2013@163.com
作者简介:厉芳，女，硕士研究生，住院医师，研究方向：神经药理。 Tel：0571-83807951 E-mail：308896208@qq.com 郑浏璞，共同第一作者，女，硕士研究生，主治医师，研究方向：器官脑保护。 Tel：13857745121E-mail：zlp3305@sina.com
基金资助:
国家自然科学基金项目（81070960）；浙江医学高等专科学校科研基金项目（2015XZBOT）

Therapeutic time window of effectiveness for idazoxan ameliorating brain damage of rats with experimental autoimmune encephalomyelitis

LI Fang ¹, ZHENG Liupu ², WENG Jianlong¹, GUO Tao ¹, SONG Xingwei ¹, ZHENG Rongyuan³

¹Department of Neurology, Xiaoshan First People's Hospital, Hangzhou 311200, Zhejiang, China; ² Department of Anesthesiology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang, China; ³ Department of Neurology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang, China

Received:2017-05-22 Revised:2017-06-15 Online:2018-01-26 Published:2018-02-07

摘要/Abstract

摘要：

目的: 探讨咪唑克生减缓大鼠实验性自身免疫性脑脊髓炎（EAE）发病的药效作用时间窗。方法: SD大鼠50只随机分为4组：空白对照组、EAE-盐水组、EAE-咪唑克生第1~7天给药组、EAE-咪唑克生第8~14天给药组。从EAE建模开始予腹腔注射咪唑克生（2 mg/kg）或等量生理盐水，至发病第15天处死。观察大鼠行为学变化、测定中枢神经系统（CNS）炎性细胞浸润和髓鞘脱失情况、小胶质细胞和星形胶质细胞的表达、炎性细胞因子白介素17（IL-17）和白介素10(IL-10)的表达。结果: 与EAE-盐水组比较，两组咪唑克生给药组大鼠发病率均下降，临床症状均减轻，潜伏期延长，但仅EAE-咪唑克生第8～14天给药组临床症状改善差异有统计学意义（P<0.05）；两组咪唑克生给药组均能减少CNS内炎性细胞浸润和髓鞘脱失程度，而仅EAE-咪唑克生第8～14天给药组炎症性髓鞘脱失病理学改变程度差异有统计学意义（P<0.05）；两组咪唑克生给药组均能减少小胶质细胞的数量和增加星形胶质细胞的数量，但仅EAE-咪唑克生第8～14天给药组减少小胶质细胞数量上有统计学差异（P<0.01）；EAE-咪唑克生第8～14天给药组中枢神经系统内IL-17表达减少，而IL-10表达增加（P<0.01）。结论: 咪唑克生对大鼠EAE的有效作用时间窗在实验大鼠免疫诱导后第8～14天（发病初始期），作用机制可能与抑制小胶质细胞的活化，抑制炎性细胞因子的表达有关。

关键词: 咪唑克生, 实验性自身免疫性脑脊髓炎, 小胶质细胞, 炎性细胞因子

Abstract:

AIM: To investigate the effective therapeutic time window of idazoxan in rats with experimental autoimmune encephalomyelitis (EAE). METHODS: Fifty SD rats were randomly divided into four groups: (1) Control group (n=8), which was given no treatment, but raised in the same condition as the experimental groups. (2) EAE-NS group (n=14), which received saline rather than idazoxan; (3) EAE-idazoxan 1-7 d group (n=14), which idazoxan was given at 2 mg/kg through i.p. (b.i.d.) from the first day of immunization to the 7th day; (4) EAE-idazoxan 8-14 d group (n=14), which received 2 mg/kg idazoxan through i.p. (b.i.d.) from the 8th day of immunization to the 14th day. Rat's behavioral changes were measured twice daily after immunizations until they were killed at 15th day after immunization to collect the cerebrum and the lumbar spinal cord. HE staining and luxol fast blue (LFB) staining was performed on the tissue sections to show tissue damage, infiltration of inflammatory cells and demyelination. Immunohistochemistry was used to determine the expression of Iba1 and GFAP in the spinal cord.RESULTS:Idazoxan improved neurological behaviours: EAE-idazoxan 8-14 d group significantly reduced the daily clinical scores and cumulative neurobehavioral scores compared with the EAE-NS group (P<0.05). Idazoxan reduced EAE histopathology: Inflammatory cell entry into the brain and spinal cord was significantly reduced in EAE-idazoxan 8-14 d group compared with the EAE-NS group (P<0.05). Demyelinationin the brain and spinal cord was also reduced in EAE-idazoxan 8-14 d group based on LFB staining when compared with EAE-NS group (P<0.05). Idazoxan reduced Iba1 expression: EAE induced significant increase in the number of Iba1 expressing cells in the brain and reduce astrocytic(GFAP) activation. Compared with the EAE-NS group, the number of Iba1 positive cells EAE-idazoxan 8-14 d Group was significantly reduced (P<0.01). ELISA analysis showed that the expressions of IL-17 (P<0.01) decreased and IL-10 increased significantly in the CNS in EAE-idazoxan 8-14 d group rats compared with EAE-NS group rats. CONCLUSION: The effective therapeutic time window for idazoxan is in the period of disease onset of EAE disease progression. The protective effect of idazoxan on EAE is associated with the attenuated microglial activation and shifting the balance of inflammatory cytokines.

Key words: idazoxan, experimental autoimmune encephalomyelitis, microglia, inflammatory cytokines

中图分类号:

R965.2

厉芳，郑浏璞，翁建龙，郭涛，宋兴伟，郑荣远. 咪唑克生减缓大鼠实验性自身免疫性脑脊髓炎脑损伤的保护作用时间窗研究[J]. 中国临床药理学与治疗学, 2018, 23(1): 1-7.

LI Fang, ZHENG Liupu, WENG Jianlong, GUO Tao, SONG Xingwei, ZHENG Rongyuan. Therapeutic time window of effectiveness for idazoxan ameliorating brain damage of rats with experimental autoimmune encephalomyelitis[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(1): 1-7.

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咪唑克生减缓大鼠实验性自身免疫性脑脊髓炎脑损伤的保护作用时间窗研究

Therapeutic time window of effectiveness for idazoxan ameliorating brain damage of rats with experimental autoimmune encephalomyelitis

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