Neuroprotective Effect and Repair Mechanism of Huangqi Glycoprotein on Mice with Experimental Autoimmune Encephalomyelitis

XING Yan-xia; LIU Bin-yu; ZHAO Yi-jin; ZHANG Li-hong; ZHANG Guang-xian; XUE Hui-qing; MA Cun-gen; ZHANG Pei-jun

doi:10.13422/j.cnki.syfjx.20182101

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Neuroprotective Effect and Repair Mechanism of Huangqi Glycoprotein on Mice with Experimental Autoimmune Encephalomyelitis

更新时间：2024-09-23

- Neuroprotective Effect and Repair Mechanism of Huangqi Glycoprotein on Mice with Experimental Autoimmune Encephalomyelitis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 24, Pages: 7-13(2018)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.20182101
  CLC： R285.5
- Published：2018
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XING Yan-xia, LIU Bin-yu, ZHAO Yi-jin, et al. Neuroprotective Effect and Repair Mechanism of Huangqi Glycoprotein on Mice with Experimental Autoimmune Encephalomyelitis[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(24): 7-13.
DOI：

XING Yan-xia, LIU Bin-yu, ZHAO Yi-jin, et al. Neuroprotective Effect and Repair Mechanism of Huangqi Glycoprotein on Mice with Experimental Autoimmune Encephalomyelitis[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(24): 7-13. DOI： 10.13422/j.cnki.syfjx.20182101.

摘要

目的：观察黄芪糖蛋白（HQGP）对实验性自身免疫性脑脊髓炎（EAE）小鼠的保护作用，并探讨其神经修复机制。方法：复制髓鞘少突胶质细胞糖蛋白35-55（MOG35-55）多肽诱导的小鼠EAE模型，20只C57BL/6雌性小鼠随机分为EAE组和HQGP组。HQGP组于免疫后第3日起，按1 mg·kg-1·d-1腹腔注射HQGP，持续给药18 d。通过五级临床症状评分和小鼠体质量变化观察HQGP对EAE小鼠的干预效应；取小鼠脊髓进行苏木精-伊红（HE）和固蓝（LFB）染色观察HQGP的神经保护作用；免疫组织化学法检测小鼠脊髓白细胞分化抗原（CD）68+巨噬细胞的浸润以及诱导型一氧化氮合酶（iNOS），微管相关蛋白2（MAP-2）和神经元核抗原（NeuN）的表达；免疫荧光组织化学双染法检测小鼠脊髓CD11b+细胞和趋化因子配体-5（CCL-5）的表达；酶联免疫吸附测定法（ELISA）检测小鼠脾脏单个核细胞（MNC）培养上清液中相关细胞因子的变化。结果： HQGP有效缓解EAE小鼠的临床症状，推迟发病时间，减轻中枢神经系统的炎性反应和髓鞘脱失；HQGP能减少小鼠脊髓CD68+巨噬细胞和CD11b+细胞的数量，抑制脊髓iNOS和趋化因子CCL-5的表达，增加MAP-2和NeuN的表达水平；HQGP能明显降低EAE小鼠肿瘤坏死因子-α（TNF-α）和白细胞介素（IL）-6的分泌，促进IL-10和γ-干扰素（IFN-γ）的分泌。结论： HQGP具有治疗EAE的潜在作用，其神经保护机制与抑制炎症反应、调控免疫细胞表达及细胞因子的分泌、减轻髓鞘脱失并促进轴突修复和神经元发育有关。

Abstract

Objective: To observe the protection of Huangqi glycoprotein (HQGP) on mice with experimental autoimmune encephalomyelitis(EAE) and further explore its mechanism of nerve repair. Method: Twenty female C57BL/6 mice were immunized subcutaneously with myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) polypeptide and randomly divided into EAE group and HQGP group.Mice in HQGP group were received intraperitoneal injection of HQGP (1 mg·kg-1·d-1) on day 3 post-immunization with continuous administration for 18 days.To observe the effect of HQGP in EAE

clinical assessment of EAE scores was evaluated and the body weight of mice was recorded every day.Spinal cords of mice were obtained for hematoxylin-eosin(HE) and Luxol fast blue(LFB) staining to observe the neuroprotective effect of HQGP.The infiltration of cluster of differentiation 68+(CD68+) macrophages and the expression of inducible nitric oxide synthase(iNOS)

microtubule-associated protein 2(MAP-2) and neuronal nuclei(NeuN) in spinal cord of mice were detected by immunohistochemistry.Double immunofluorescence histochemistry was used to detect the expression of CD11b+ cells and chemokine ligand-5(CCL-5) in spinal cord of mice.The levels of cytokines in the supernatant of cultured splenic suspension of mononuclear cells(MNCs) were determined by enzyme-linked immunosorbent assay(ELISA). Result: HQGP delayed onset and improved the clinical symptoms of EAE

accompanied by inhibiting inflammation and alleviating demyelination in the spinal cord.Compared with EAE group

HQGP reduced the infiltration of CD68+ macrophages and CD11b+ cells

inhibited the expression of iNOS and CCL-5 and increased the expression of MAP-2 and NeuN in the spinal cord.Further studies showed that HQGP could reduce the production of tumor necrosis factor(TNF)-α and interleukin(IL)-6

increase the level of IL-10 and γ-interferon(IFN-γ). Conclusion: HQGP has the potential role for the treatment of EAE and the mechanism is related to anti-inflammation

regulation of immune cell expression and cytokine secretion

reduction of demyelination and promotion of axon and neuron repair.

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