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菖蒲挥发油通过调节多巴胺能和谷氨酸能系统影响抽动障碍大鼠的突触可塑性。

Volatile oil from Acori graminei Rhizoma affected the synaptic plasticity of rats with tic disorders by modulating dopaminergic and glutamatergic systems.

机构信息

Medical College, Hexi University, Zhangye, Gansu, China.

Clinical College of Chinese Medicine, Gansu University of Chinese Medicine, Lanzhou, Gansu, China.

出版信息

J Ethnopharmacol. 2024 Dec 5;335:118676. doi: 10.1016/j.jep.2024.118676. Epub 2024 Aug 13.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Acori graminei Rhizoma is a commonly used traditional Chinese medicine for treating TD, with its main component being calamus volatile oil. Volatile Oil from Acori graminei Rhizoma (VOA)can protect nerve cells and alleviate learning and memory disorders. However, the mechanism of anti-tic of VOA is still unclear.

AIM OF THE STUDY

We aimed to explore the effects of Volatile Oil from Acori Tatarinowii Rhizoma (VOA) on striatal dopaminergic and glutamatergic systems and synaptic plasticity of rats with Tic Disorder (TD), as well as its pharmaceutical mechanism against TD.

MATERIALS AND METHODS

This study involved 48 (three-week-old) Sprague Dawley (SD) rats, which were randomly divided into two primary groups: Control (8) and TD (40). Rats in the TD group were injected intraperitoneally with 3,3-iminodipropionitrile (IDPN) to construct the TD rat model. They were divided into five subgroups: Model, Tiapride, VOA-high, VOA-medium, and VOA-low (N = 8). After modeling, VOA was administrated to rats in the VOA groups through gavage (once/day for four consecutive weeks), while rats in the blank control and model groups received normal saline of the same volume. The animals' behavioral changes were reflected using the stereotypic and motor behavior scores. After interferences, patterns of striatal neurons and the density of dendritic spines were investigated using H&E and Golgi staining, and the ultrastructure of striatal synapses was examined using Transmission Electron Microscopy (TEM). Furthermore, Ca content was determined using the Ca detector, and Dopamine (DA) and Glutamate (GLU) contents in serum and striatum were detected through ELISA. Finally, DRD1, DRD2, AMPAR1, NMPAR1, DAT, VMAT2, CAMKⅡ, and CREB expression in the striatum was detected using Quantitative real-time PCR (qRT-PCR), Western Blotting (WB) and Immunohistochemical (IHC) methods.

RESULTS

Compared to rats in the blank control and model groups, rats in the VOA groups showed lower stereotypic behavior scores. Furthermore, rats in the VOA groups exhibited relieved, neuron damage and increased quantities of neuronal dendrites and dendritic spines Additionally, based on TEM images show that, the VOA groups showed a clear synaptic structure and increased amounts of postsynaptic dense substances and synaptic vesicles. The VOA groups also exhibited reduced Ca contents, and upregulation of DRD1, DRD2, DAT, AMPAR1, and NMPAR1 and downregulation of VMAT-2, CAMKⅡ, and CREB in the striatum.

CONCLUSIONS

In summary, VOA could influence synaptic plasticity by tuning the dopaminergic and glutamatergic systems, thus relieving TD.

摘要

ethnopharmacological 相关性:菖蒲根茎是一种常用于治疗抽动障碍的传统中药,其主要成分是菖蒲挥发油。菖蒲根茎挥发油(VOA)可以保护神经细胞,减轻学习和记忆障碍。然而,VOA 的抗抽搐机制尚不清楚。

目的

本研究旨在探讨菖蒲根茎挥发油(VOA)对抽动障碍(TD)大鼠纹状体多巴胺能和谷氨酸能系统及突触可塑性的影响,以及其治疗 TD 的药物机制。

材料和方法

本研究共涉及 48 只(3 周龄)斯普拉格-道利(SD)大鼠,随机分为两组:对照组(8 只)和 TD 组(40 只)。TD 组大鼠腹腔注射 3,3-亚氨基二丙腈(IDPN)构建 TD 大鼠模型。它们分为五组:模型组、泰必利组、VOA-高剂量组、VOA-中剂量组和 VOA-低剂量组(每组 8 只)。建模后,通过灌胃给予 VOA 组大鼠 VOA(连续四周,每天一次),空白对照组和模型组给予相同体积的生理盐水。采用刻板行为和运动行为评分反映动物行为变化。干扰后,采用 H&E 和高尔基染色观察纹状体神经元形态和树突棘密度,采用透射电镜(TEM)观察纹状体突触超微结构。此外,采用钙探测器测定钙含量,采用 ELISA 法检测血清和纹状体中多巴胺(DA)和谷氨酸(GLU)含量。最后,采用实时定量 PCR(qRT-PCR)、Western blot(WB)和免疫组化(IHC)方法检测纹状体中 DRD1、DRD2、AMPA1、NMPAR1、DAT、VMAT2、CAMKⅡ和 CREB 的表达。

结果

与空白对照组和模型组大鼠相比,VOA 组大鼠刻板行为评分较低。此外,VOA 组大鼠神经元损伤减轻,神经元数量增加,树突数量和树突棘数量增加。此外,TEM 图像显示,VOA 组突触结构清晰,突触后致密物质和突触小泡数量增加。VOA 组钙含量降低,纹状体中 DRD1、DRD2、DAT、AMPA1、NMPAR1 表达上调,VMAT-2、CAMKⅡ、CREB 表达下调。

结论

综上所述,VOA 可通过调节多巴胺能和谷氨酸能系统影响突触可塑性,从而缓解 TD。

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