本文選題：甲基苯丙胺 + 微小RNA��； 參考：《寧波大學》2017年碩士論文

【摘要】：目的:目前甲基苯丙胺(METH)濫用依然是全球性的公共健康問題。長期使用甲基苯丙胺會導致多種精神疾病,但甲基苯丙胺成癮神經(jīng)生物學機制仍未明確。本文從甲基苯丙胺成癮中的microRNA入手,篩選出靶向中樞血管緊張素系統(tǒng)的microRNAs,通過調(diào)節(jié)microRNA來探究中樞血管緊張素系統(tǒng)(RAS)在甲基苯丙胺成癮中的潛在作用。方法:1、將甲基苯丙胺靜脈自身主動給藥組(甲基苯丙胺主動組)、甲基苯丙胺靜脈被動給藥組(甲基苯丙胺被動組)和生理鹽水靜脈主動給藥組(生理鹽水組)的大鼠伏隔核進行microRNA microarray,找出異常表達的microRNA;基于GO分析與KEGG分析,對中樞血管經(jīng)張素系統(tǒng)有關的microRNA進行篩選;2、通過微注射慢病毒在大鼠腦區(qū)-伏隔核中過表達miR-219a-5p,分別通過固定頻率(FR)和累進頻率(PR)來檢驗miR-219a-5p過表達后對大鼠強迫給藥和給藥動機的調(diào)控作用;3、通過微注射慢病毒在大鼠腦區(qū)-伏隔核中過表達miR-219a-5p,探索miR-219a-5p在甲基苯丙胺復吸中的調(diào)控作用;4、通過酶聯(lián)免疫吸附測定法來測定醛固酮合成和分泌通路中血管緊張素II受體1(AT1)、磷脂酰肌醇磷脂酶(PLCβ)和cAMP反應元件結合蛋白(CREB)等蛋白在甲基苯丙胺自身給藥動物中是否有顯著變化;5、根據(jù)實驗四的驗證,對實驗二和實驗三中的大鼠伏隔核腦區(qū),在miR-219a-5p過表達后,檢測主要蛋白的表達變化。結果:1、基于甲基苯丙胺主動組和甲基苯丙胺被動組以及甲基苯丙胺主動組和生理鹽水組分別進行比較,篩選出顯著變化的靶向中樞血管緊張素系統(tǒng)的microRNAs,最后,根據(jù)相對表達倍數(shù)篩選出研究目標miR-219a-5p;2、miR-219a-5p過表達后,對于固定頻率給藥,在0.025mg/kg的劑量中,LV1-miR-219a-5p組與LV1CN組之間沒有差異,而在0.05mg/kg、0.075mg/kg和0.1mg/kg中,LV1-miR-219a-5p組相對于LV1CN組給藥次數(shù)顯著下降。對于累進頻率給藥,在0.025mg/kg的劑量中,LV1-miR-219a-5p組與LV1CN組之間沒有差異,而在0.05mg/kg、0.075mg/kg和0.1mg/kg中,LV1-miR-219a-5p組相對于LV1CN組給藥次數(shù)顯著下降;3、miR-219a-5p過表達后,無論是線索誘導還是藥物誘導,LV1-miR-219a-5p組相對于LV1CN組有效鼻觸數(shù)顯著下降;4、通過酶聯(lián)免疫吸附測定法(ELISA),AT1、PLCβ和CREB等蛋白表達水平在甲基苯丙胺成癮中顯著上升;5、通過免疫印跡法測定實驗二和實驗三中的大鼠伏隔核腦區(qū)AT1、PLCβ和CREB表達情況,可以發(fā)現(xiàn)LV1-miR-219a-5p組相對于LV1CN組蛋白的表達量顯著下降。結論:我們的研究結果表明,大鼠伏隔核中的miR-219a-5p在中樞腎素-血管緊張素-醛固酮系統(tǒng)(RAAS)的重要作用,揭示了調(diào)控甲基苯丙胺成癮的分子機制。
[Abstract]:Objective: the abuse of methamphetamine methamphetamine (METHH) is still a global public health problem. Long-term use of methamphetamine can lead to many mental disorders, but the neurobiological mechanism of methamphetamine addiction remains unclear. Starting with microRNA in methamphetamine addiction, microRNAs-targeted central angiotensin system was screened to explore the potential role of central angiotensin system in methamphetamine addiction by regulating microRNA. Methods: 1, methamphetamine group (methamphetamine active group), methamphetamine group (methamphetamine passive group) and saline group (saline group) were divided into three groups: methamphetamine self-administered group (methamphetamine active group), methamphetamine group (methamphetamine passive group) and saline group (saline group). MicroRNA microRNAs were detected in the nucleus accumbens of rats, based on go analysis and KEGG analysis. The microRNA related to the tensin system in central blood vessels was screened. The overexpression of miR-219a-5p in rat brain region and nucleus accumbens was detected by microinjection of lentivirus, and forced injection of miR-219a-5p was performed by fixed frequency and progressive frequency. By microinjection of lentivirus into rat brain region and nucleus accumbens to investigate the regulatory role of miR-219a-5p in methamphetamine relapse, aldosterone was determined by enzyme-linked immunosorbent assay (enzyme-linked immunosorbent assay). Whether angiotensin II receptor 1 (AT1), phosphatidylinositol phospholipase 尾 (PLC- 尾) and cAMP response element binding protein (CREBB) have significantly changed in methamphetamine self-administered animals. After overexpression of miR-219a-5p in the nucleus accumbens of rats in experiment two and three, the expression of major proteins was detected. Results: 1, based on the comparison between the active methamphetamine group and the methamphetamine passive group, and the methamphetamine active group and the normal saline group, the microRNAss targeting the central angiotensin system were screened out. After over expression of miR-219a-5pn2miR-219a-5p, there was no difference between LV1-miR-219a-5p group and LV1CN group in the dose of 0.025mg/kg, but in 0.05mg / kg of LV1-miR-219a-5p group and 0.1mg/kg group, LV1-miR-219a-5p group was significantly lower than that of LV1CN group. For progressive frequency administration, there was no difference between LV1-miR-219a-5p group and LV1CN group in the dose of 0.025mg/kg. However, LV1-miR-219a-5p group was significantly lower than LV1CN group in the frequency of administration of LV1-miR-219a-5p in 0.05 mg / kg and 0.1mg/kg 0.075 mg / kg, compared with that in LV1CN group, the expression of LV1-miR-219a-5p was significantly lower than that of LV1CN group, and the expression of LV1-miR-219a-5p was significantly lower than that of LV1CN group. Both cue-induced and drug-induced LV1-miR-219a-5p groups significantly decreased the effective nasal contacts compared with those of LV1CN group. The expression levels of ELISAA AT1 CREB 尾 and CREB in methamphetamine addiction were significantly increased by Elisa. The expression of PLC 尾 and CREB in the nucleus accumbens of rats in experiment two and three were measured by trace method. It was found that the expression of histone in LV1-miR-219a-5p group was significantly lower than that in LV1CN group. Conclusion: our results suggest that miR-219a-5p in rat nucleus accumbens plays an important role in central renin-angiotensin-aldosterone system, which reveals the molecular mechanism of regulating methamphetamine addiction.
【學位授予單位】：寧波大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R749.64

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1 潘儉;miR-219a-5p在甲基苯丙胺成癮中的作用[D];寧波大學;2017年

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本文編號：1868266