青蒿琥酯对实验性自身免疫性脑脊髓炎小鼠的神经保护作用及细胞自噬的影响
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(西南医科大学附属医院神经内科,四川泸州 646000)

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R-33

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Neuroprotective effect of artesunate on experimental autoimmune encephalomyelitis and on autophagy in mice
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(Department of Neurology, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, China)

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    摘要:

    目的 探讨青蒿琥酯对实验性自身免疫性脑脊髓炎(EAE)小鼠神经保护作用及细胞自噬影响?方法 选择48 只雌性C57BL/6 小鼠随机分为空白组?模型组?青蒿琥酯低剂量组?高剂量组共4 组,每组12 只?采用MOG35-55 多肽制备EAE 模型,青蒿琥酯低剂量组?高剂量组分别予以青蒿琥酯(10 mg/ (kg·d),50 mg/ (kg·d))腹腔注射,连续10 d,观察小鼠发病情况?脑组织行luxol fast blue(LFB)染色观察脱髓鞘情况,采用Westernblot 法检测自噬相关蛋白LC3-I?LC3-II 的表达?结果 空白组小鼠均未发病,模型组?青蒿琥酯各剂量组发病小鼠出现不同程度的尾部下垂?行走不稳?后肢无力等?青蒿琥酯各剂量组较模型组相比,发病潜伏期?高峰期延迟?神经功能评分减低,高剂量组较低剂量组作用明显( P <0. 05),低剂量组和高剂量组发病高峰期比较无统计学意义( P >0. 05)?②LFB 染色见模型组脑组织髓鞘排列疏松?紊乱?低染,青蒿琥酯各剂量组髓鞘染色情况好转?③Western blot 检测模型组与空白组相比, LC3-I?LC3-II 条带光密度值及LC3-II/ LC3-I 比值均增加( P <0. 01);青蒿琥酯各剂量组较模型组相比,LC3I?LC3II 条带光密度值及 LC3-II/ LC3-I 比值均减小( P <0. 01),高剂量组较低剂量组作用明显( P <0. 05)?结论 青蒿琥酯对EAE 小鼠具有神经保护作用,减轻脑组织脱髓鞘情况,且作用机制可能与通过下调LC3-I?LC3-II 和LC3-II/ LC3-I 减轻细胞自噬有关?

    Abstract:

    Objective To investigate the neuroprotective effects of artesunate on experimental autoimmuneencephalomyelitis (EAE) and on autophagy in mice. Methods Forty-eight female C57BL/6 mice were randomly dividedinto four groups: control group, model group, and artesunate low- and high-dose groups, with 12 mice in each group. TheEAE model was induced by MOG35-55 peptide. The mice in the low- and high-dose groups were intraperitoneally injectedwith artesunate (10 or 50 mg/ (kg·d)) for 10 consecutive days. The symptoms of the mice in each group were observed.Demyelination lesions in the brain tissues were observed by luxol fast blue (LFB) staining. The expression levels ofautophagy protein markers LC3-I and LC3-II were detected through western blot analysis. Results ①The mice in thecontrol group did not develop neurological symptoms. The mice in the model group and artesunate groups developed tovarying degrees of gait instability, hindlimb weakness, and paralysis. Compared with the model group, the latent period andpeak period were delayed and neurofunctional deficiency scores were decreased in the artesunate groups. The effects in thehigh-dose group were more pronounced than those in the low-dose group ( P < 0. 05). There was no significant difference inpeak period between the low- and high-dose artesunate groups ( P > 0. 05). ② LFB staining showed that the myelin sheathof brain tissue in the model group was loose, disordered, and had low staining intensity, while these findings were improvedin the artesunate groups. ③ Western blot analysis showed that the optical density values of LC3-I, LC3-II, and LC3-II/LC3-I in the model group were higher than those in the control group ( P < 0. 01). These values were lower in the artesunategroups than in the model group ( P < 0. 01), and the findings in the high-dose artesunate group were more pronounced thanthose in the low-dose artesunate group ( P < 0. 05). Conclusions Artesunate has neuroprotective effects on EAE mice andcan reduce demyelination in brain tissue. The mechanism involved may be related to the alleviation of autophagy by downregulation of LC3-I, LC3-II, and LC3-II/ LC3-I expressions.

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罗思维,李作孝.青蒿琥酯对实验性自身免疫性脑脊髓炎小鼠的神经保护作用及细胞自噬的影响[J].中国比较医学杂志,2019,29(4):65~68.

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  • 收稿日期:2018-09-16
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  • 在线发布日期: 2019-05-10
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