不同剂量天麻素对甲基苯丙胺依赖CPP大鼠海马中TLR4/MyD88/NF-κB炎症信号通路的影响

朱婷娜; 曹媛媛; 张园; 刘鹏亮; 王一航; 吴亚梅; 李利华; 赵永娜; 洪仕君

doi:10.12259/j.issn.2095-610X.S20230226

不同剂量天麻素对甲基苯丙胺依赖CPP大鼠海马中TLR4/MyD88/NF-κB炎症信号通路的影响

doi: 10.12259/j.issn.2095-610X.S20230226

朱婷娜^{1, 4},
曹媛媛¹,
张园¹,
刘鹏亮²,
王一航²,
吴亚梅¹,
李利华²,
赵永娜^{1, 3, ,},
洪仕君^2, ,

1.
昆明医科大学药学院暨云南省天然药物药理重点实验室
2.
法医学院
3.
国际教育学院，云南　昆明　650500
4.
云南省食品药品监督检验研究院，云南昆明　650106

基金项目: 国家自然科学基金资助项目（81760337 ）；云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目［2019FE001（-163）］

详细信息

作者简介:
朱婷娜（1994～），女，云南宣威人，硕士，主要从事神经药理学研究工作

通讯作者:
赵永娜，E-mail：mszhaoyn@163.com

洪仕君，E-mail：kmhongshijun@126.com

中图分类号: R964
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- 被引次数: 0
出版历程
- 收稿日期: 2022-12-19
- 网络出版日期: 2023-02-01
- 刊出日期: 2023-02-25

Effects of Different Doses of Gastrodin on TLR4/MyD88/NF-κB Inflammatory Signal Pathway in Hippocampus of Methamphetamine Dependent CPP Rats

1.
School of Pharmaceutical Sciences & Yunnan Key Laboratory of Pharmacology for Natural Products
2.
School of Forensic Medicine
3.
School of International Education，Kunming Medical University，Kunming，Yunnan 650500
4.
Yunnan Institute for Food and Drug Contral， Kunming Yunnan 650106，China

摘要

摘要: 目的探讨不同剂量的天麻素（Gastrodin，Gas）对甲基苯丙胺（Methamphetamine，Meth）依赖条件性位置偏爱（Conditioned place preference，CPP）大鼠海马中TLR4/MyD88/NF-κB通路的作用。方法建立Meth（10 mg/kg，ip，14 d）依赖大鼠CPP模型，采用低中高剂量的天麻素（10、30、100 mg/kg）干预14 d后，采用Western blotting和RT-qPCR方法检测TLR4/MyD88/NF-κB信号通路中关键因子的蛋白和mRNA在海马中的表达水平。结果天麻素干预后TLR4、MyD88、NF-κB p65、TRAF6的蛋白和mRNA表达呈剂量依赖性降低（P < 0.001、P < 0.01或P < 0.05）、p-NF-κB p65的蛋白表达呈剂量依赖性降低（P < 0.01或P < 0.05）、IκB-α的蛋白和mRNA表达呈剂量依赖性升高（P < 0.01或P < 0.05），天麻素干预后p-IκB-α的蛋白表达降低（P < 0.01）。结论天麻素干预对甲基苯丙胺依赖CPP大鼠海马的神经炎症具有保护作用，且与TLR4/MyD88/NF-κB信号通路密切相关。
- 甲基苯丙胺依赖 /
- 条件性位置偏爱 /
- 海马 /
- 天麻素 /
- TLR4/MyD88/NF-κB信号转导通路
Abstract: Objective To investigate the effects of different dosages of gastrodin onTLR4/ MyD88 / NF-κB signal pathway in the hippocampus of methamphetamine （meth）dependence rats. Methods The conditioned place preference （CPP） model of meth was established, meth （10mg/kg） was administered （i.p, 14 d）, and then, different dosages （10, 30, 100 mg/kg） of gastrodin were used to intervene in 14 days. Western blotting and RT-qPCR were used to explore the expression level of protein and mRNA of key factors in TLR4/ MyD88 / NF-κB signal pathway in hippocampus. Results Gastrodin decreased the protein and mRNA expression of TLR4, MyD88, TRAF6, NF-κBp65 in a dose-dependent manner （P < 0.001, P < 0.01 or P < 0.05）, and the protein expression of p-NF-κB p65 was reduced with showing a significant dose-response relationship （P < 0.01 or P < 0.05）. Gastrodin increased the protein and mRNA expression of IκB-α with showing a significant dose-response relationship （P < 0.01 or P < 0.05）. Gastrodin reduced the protein expression of p-IκB-α （P < 0.01）. Conclusion Gastrodin intervention has a protective effect against hippocampal neuroinflammation in methamphetamine-dependent CPP rats, and is closely related to the function of TLR4/MyD88/NF-κB signal pathway.
- Methamphetamine dependence /
- Conditioned place preference /
- Hippocampus /
- Gastrodin /
- TLR4/MyD88/NF-κB signal transduction pathway

HTML全文

图 1 不同剂量天麻素对甲基苯丙胺依赖CPP大鼠海马TLR4、MyD88、TRAF6、p-IκB-α、IκB-α、NF-κB p65、p-NF-κB p65蛋白表达的影响

A～G：TLR4、MyD88、TRAF6、p-IκB-α、IκB-α、p-NF-κB p65、NF-κB p65蛋白的定量分析。与对照组比较，^*P < 0.05；^**P < 0.01；与Meth组比较，^#P < 0.05，^##P < 0.01。

Figure 1. The effect of different doses of gastrodin on the expression of TLR4，MyD88，TRAF6，p-IκB-α，IκB-α，p-NF-κB p65，NF-κB p65 proteins in the hippocampus of methamphetamine-dependent rats

下载: 全尺寸图片幻灯片

图 2 不同剂量天麻素对Meth依赖大鼠海马TLR4、MyD88、TRAF6、IκB-α、NF-κB p65 mRNA表达的影响

A-E：TLR4、MyD88、TRAF6、IκB-α、NF-κB p65 mRNA的定量分析。与对照组比较，^**P < 0.01；^***P < 0.001。与Meth组比较，^#P < 0.05，^##P < 0.01，^###P < 0.001。

Figure 2. The effect of different doses of gastrodin on the mRNA expression of TLR4，MyD88，TRAF6，IκB-α，NF-κB p65 in the hippocampus of meth-dependent rats

下载: 全尺寸图片幻灯片

表 1 引物序列

Table 1. Primer Sequences

基因名称	引物序列	Tm
TLR4	上游引物：5′-TGCCTGAGACCAGGAAGCTTG 3′ 下游引物：5′-CTTAAGATCTTCAGGGGGTTG3′	54 ℃
MyD88	上游引物：5′-TGAGAAAAGGTGTCGTCGCA3′ 下游引物：5′-GGGTCCAGAACCAGGACTTG′	54 ℃
TRAF6	上游引物：5′-GCCCATGCCGTATGAAGAGA 3′ 下游引物：5′-CGTGACAGCCAAACACACTG3′	54 ℃
NF-κB p65	上游引物：5′-GAGACCTGGAGCAAGCCATT3′ 下游引物：5′-AGTTCCGGTTTACTCGGCAG3′	54 ℃
IκB -α	上游引物：5′-GAATCCTGACCTGGTCTCGC′ 下游引物：5′-CACAGTCATCGTAGGGCAACT3′	55 ℃
β-actin	上游引物：5′-AGACAGCCGCATCTTGT-3′ 下游引物：5′-CTTGCCGTGGGTAGAGTCAT-3′	55 ℃

下载: 导出CSV

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