Effects of Different Doses of Gastrodin on TLR4/MyD88/NF-κB Inflammatory Signal Pathway in Hippocampus of Methamphetamine Dependent CPP Rats
-
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。 -
关键词:
- 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. -
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
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
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
-
[1] Jayanthi S,Daiwile A P,Cadet J L. Neurotoxicity of methamphetamine: Main effects and mechanisms[J]. Experimental Neurology,2021,344(10):113795. [2] Mizoguchi H,Yamada K. Methamphetamine use causes cognitive impairment and altered decision-making[J]. Neurochemistry international,2019,124(3):106-113. [3] Kang E,Wen Z,Song H,et al. Adult Neurogenesis and Psychiatric Disorders[J]. Cold Spring Harbor Perspectives in Biology,2016,8(9):a019026. doi: 10.1101/cshperspect.a019026 [4] 赵敏. 物质依赖与精神疾病的共病问题[J]. 中国药物滥用防治杂志,2013,19(4):187-189. [5] Suleymanova E M. Behavioral comorbidities of epilepsy and neuroinflammation: Evidence from experimental and clinical studies[J]. Epilepsy & Behavior,2021,117(4):107869. [6] Fernández-Arjona María Del Mar,Grondona JM,Fernández-Llebrez P,et al. Microglial activation by microbial neuraminidase through TLR2 and TLR4 receptors[J]. Journal of Neuroinflammation,2019,16(1):1-14. doi: 10.1186/s12974-018-1391-2 [7] Vargas A M,Rivera-Rodriguze DE,Martinez LR. Methamphetamine alters the TLR4 signaling pathway,NF-κB activation,and pro-inflammatory cytokine production in LPS-challenged NR-9460 microglia-like cells[J]. Molecular Immunology,2020,121(5):159-166. [8] Xie X L,Zhou W T,Zhang K K,et al. METH-Induced Neurotoxicity Is Alleviated by Lactulose Pretreatment Through Suppressing Oxidative Stress and Neuroinflammation in Rat Striatum[J]. Frontiers in Neuroscience,2018,12(11):802. [9] 房尚萍,李海源,丁磊,等. 通过调节NF-κB通路中的IκB-α抑制乳腺癌的研究进展[J]. 锦州医科大学学报,2021,42(1):104-108. [10] 朱婷娜,刘鹏亮,董文娟,等. 不同剂量天麻素对甲基苯丙胺依赖大鼠条件位置偏爱及海马小胶质细胞激活的影响[J]. 昆明医科大学学报,2021,42(5):6-11. doi: 10.12259/j.issn.2095-610X.S20210502 [11] Gonçalves J,Baptista S,Martins T,et al. Methamphetamine-induced neuroinflammation and neuronal dysfunction in the mice hippocampus:Preventive effect of indomethacin[J]. The European Journal of Neuroscience,2010,31(2):315-326. doi: 10.1111/j.1460-9568.2009.07059.x [12] Mahmoudiasl G R,Abbaszadeh H A,Rezaei-Tavirani M,et al. Nod-like receptor protein 3 and nod-like receptor protein 1 inflammasome activation in the hippocampal region of postmortem methamphetamine chronic user[J]. Bratislavske Lekarske Llisty,2019,120(10):769-776. [13] Gonçalves J,Leitão R A,Higuera-Matas A,et al. Extended-access methamphetamine self-administration elicits neuroinflammatory response along with blood-brain barrier breakdown[J]. Brain,Behavior,and Immunity,2017,62(4):306-317. [14] Andoh M,Ikegaya Y,Koyama R. Microglia modulate the structure and function of the hippocampus after early-life seizures[J]. Journal of Pharmacological Sciences,2020,144(4):212-217. doi: 10.1016/j.jphs.2020.09.003 [15] Mckendrick G,Graziane N M. Drug-Induced Conditioned Place Preference and Its Practical Use in Substance Use Disorder Research[J]. Frontiers in Behavioral Neuroscience,2020,14(1):582147. [16] Bardo M T,Bevins R A. Conditioned place preference: what does it add to our preclinical understanding of drug reward?[J]. Psychopharmacology,2000,153(1):31-43. doi: 10.1007/s002130000569 [17] Green T A,Bardo M T. Opposite regulation of conditioned place preference and intravenous drug self-administration in rodent models: Motivational and non-motivational examples[J]. Neuroscience and Biobehavioral Reviews,2020,116(9):89-98. [18] Qu H,Liu R,Chen J,et al. Aerobic Exercise Inhibits CUMS-Depressed Mice Hippocampal Inflammatory Response via Activating Hippocampal miR-223/TLR4/MyD88-NF-κB Pathway[J]. International Journal of Environmental Research and Public Health,2020,17(8):2676. doi: 10.3390/ijerph17082676 [19] Cui Y,Wang Y,Zhao D,et al. Loganin prevents BV-2 microglia cells from Aβ(1-42) -induced inflammation via regulating TLR4/TRAF6/NF-κB axis[J]. Cell Biology International,2018,42(12):1632-1642. doi: 10.1002/cbin.11060 [20] Garmabi B,Mohaddes R,Rezvani F,et al. Erythropoietin improve spatial memory impairment following methamphetamine neurotoxicity by inhibition of apoptosis,oxidative stress and neuroinflammation in CA1 area of hippocampus[J]. Journal of Chemical Neuroanatomy,2022,124(6):102137. [21] Re GF,Li H,Yang JQ,et al. Exercise modulates central and peripheral inflammatory responses and ameliorates methamphetamine-induced anxiety-like symptoms in mice[J]. Frontiers in Molecular Neuroscience,2022,15(8):955799. [22] Du SH,Qiao DF,Chen CX,et al. Toll-Like Receptor 4 Mediates Methamphetamine-Induced Neuroinflammation through Caspase-11 Signaling Pathway in Astrocytes[J]. Frontiers in Molecular Neuroscience,2017,10(12):409. [23] Wang X,Chen L,Xu Y,et al. Gastrodin alleviates perioperative neurocognitive dysfunction of aged mice by suppressing neuroinflammation[J]. European Journal of Pharmacology,2021,892(3):173734. [24] Li S,Bian L,Fu X,et al. Gastrodin pretreatment alleviates rat brain injury caused by cerebral ischemic-reperfusion[J]. Brain Research,2019,1712(11):207-216. [25] Ye T,Meng X,Wang R,et al. Gastrodin Alleviates Cognitive Dysfunction and Depressive-Like Behaviors by Inhibiting ER Stress and NLRP3 Inflammasome Activation in db/db Mice[J]. International Journal of Molecular Sciences,2018,19(12):3977. doi: 10.3390/ijms19123977 [26] 刘英杰,姜茜,张胜娜. 天麻素对毛果芸香碱诱发的癫痫大鼠TLR4/NF-κB信号通路影响的研究[J]. 新中医,2020,52(4):1-6. doi: 10.13457/j.cnki.jncm.2020.04.001 [27] Mei Z,Huang B,Qian X,et al. Gastrodin improves preeclampsia-induced cell apoptosis by regulation of TLR4/NF-κB in rats[J]. Food Science & Nutrition,2020,8(2):820-829. [28] 文欢,张大燕,王伟,等. 天麻素对糖氧剥夺再复供皮层神经细胞NF-κB炎症级联信号通路表达的影响[J]. 中国实验方剂学杂志,2017,23(21):104-111. doi: 10.13422/j.cnki.syfjx.2017210104 [29] Zheng X,Gong T,Tang C,et al. Gastrodin improves neuroinflammation-induced cognitive dysfunction in rats by regulating NLRP3 inflammasome[J]. BMC Anesthesiology,2022,22(1):1-13. doi: 10.1186/s12871-021-01532-1 [30] 薛凤麟,洪仕君,曾晓峰,等. 天麻素注射液通过调节背侧纹状体miR-30a表达改善大鼠甲基苯丙胺成瘾的实验研究[J]. 中药药理与临床,2020,36(1):85-89. doi: 10.13412/j.cnki.zyyl.2020.01.009 [31] 薛凤麟,洪仕君,曾晓峰,等. 基于nNOS途径的天麻素注射液改善甲基苯丙胺诱导大鼠神经毒性损伤的作用机制研究[J]. 中国药房,2020,31(10):1171-1178. doi: 10.6039/j.issn.1001-0408.2020.10.04 期刊类型引用(2)
1. 张玲霞. 低剂量冠状动脉CT结合基于模型的高级迭代重建技术在PCI术后复查中的应用. 影像研究与医学应用. 2023(13): 93-95 . 
百度学术2. 王耀东. ADMIRE低管电压CT扫描对上腹部平扫患者辐射剂量及图像质量的影响. 中国民康医学. 2022(02): 134-137 . 百度学术其他类型引用(1)
-
点击查看大图
图(2) / 表(1)
计量
- 文章访问数: 4229
- HTML全文浏览量: 1914
- PDF下载量: 7
- 被引次数: 3
