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葛根素上调血管性痴呆大鼠海马甲基化 CpG 结合蛋白 2 的磷酸化。

Puerarin Up-regulates Methyl-CpG Binding Protein 2 Phosphorylation in Hippocampus of Vascular Dementia Rats.

机构信息

Department of Neurology, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710004, China.

出版信息

Chin J Integr Med. 2018 May;24(5):372-377. doi: 10.1007/s11655-018-2822-0. Epub 2018 Jan 9.

Abstract

OBJECTIVE

To observe the effect of puerarin on methyl-CpG binding protein 2 (MeCP2) phosphorylation (pMeCP2) in the hippocampus of a rat model of vascular dementia (VD).

METHODS

Thirty-six healthy Sprague-Dawley rats were randomly assigned to the sham-operated group, dementia group and puerarintreated group using a random number table (n=12 per group). The modifified permanent bilateral common carotid artery occlusion method was used to establish the VD model. The sham-operated and dementia groups were given 2 mL/d of saline, while the puerarin-treated group was given 100 mg/(kg•d) of puerarin for 17 days. The learning and memory abilities were evaluated by the Morris water maze test. Hematoxylin-eosin staining, immunohistochemical (IHC) staining and Western blot analysis were carried out to observe changes in neuron morphology and in level of pMeCP2 in the hippocampus, respectively.

RESULTS

The morphologies of rat hippocampal neurons in the puerarintreated group were markedly improved compared with the dementia group. The escape latency of the dementia group was significantly longer than the sham-operated group (P<0.05), while the puerarin-treated group was obviously shorter than the dementia group (P<0.05). Cross-platform times of the dementia group were signifificantly decreased compared with the sham-operated group (P<0.05), while the puerarin-treated group was obviously increased compared with the dementia group (P<0.05). IHC staining showed no significant difference in the number of MeCP2 positive cells among 3 groups (P>0.05). The number of pMeCP2 positive cells in the CA1 region of hippocampus in the dementia group was signifificantly increased compared with the sham-operated group, and the puerarin-treated group was signifificantly increased compared with the dementia group (both P<0.05). Western blot analysis showed no signifificant difference of MeCP2 expression among 3 groups (P>0.05). The expression of pMeCP2 in the dementia group was signifificantly increased compared with the sham-operated group, while it in the puerarin-treated group was signifificantly increased compared with the dementia group (P<0.05).

CONCLUSION

Puerarin could play a role in the protection of nerve cells through up-regulating pMeCP2 in the hippocampus, improving neuron morphologies, and enhancing learning and memory ablities in a rat model of VD.

摘要

目的

观察葛根素对血管性痴呆(VD)大鼠模型海马中甲基化CpG 结合蛋白 2(MeCP2)磷酸化(pMeCP2)的影响。

方法

36 只健康的 Sprague-Dawley 大鼠采用随机数字表法(n=12 只/组)分为假手术组、痴呆组和葛根素治疗组。采用改良的永久性双侧颈总动脉闭塞法建立 VD 模型。假手术组和痴呆组给予 2 mL/d 生理盐水,葛根素治疗组给予 100 mg/(kg·d)葛根素治疗 17 天。采用 Morris 水迷宫试验评估学习记忆能力。通过苏木精-伊红染色、免疫组织化学(IHC)染色和 Western blot 分析观察海马神经元形态和 pMeCP2 水平的变化。

结果

与痴呆组相比,葛根素治疗组大鼠海马神经元形态明显改善。痴呆组的逃避潜伏期明显长于假手术组(P<0.05),而葛根素治疗组明显短于痴呆组(P<0.05)。痴呆组的穿越平台次数明显少于假手术组(P<0.05),而葛根素治疗组明显多于痴呆组(P<0.05)。IHC 染色显示 3 组 MeCP2 阳性细胞数无明显差异(P>0.05)。痴呆组海马 CA1 区 pMeCP2 阳性细胞数明显多于假手术组,葛根素治疗组明显多于痴呆组(均 P<0.05)。Western blot 分析显示 3 组 MeCP2 表达无明显差异(P>0.05)。痴呆组 pMeCP2 表达明显高于假手术组,葛根素治疗组明显高于痴呆组(P<0.05)。

结论

葛根素可能通过上调海马 pMeCP2 发挥神经保护作用,改善神经元形态,增强血管性痴呆大鼠学习记忆能力。

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