氣體信號分子硫化氫對慢性不可預(yù)知性應(yīng)激所致大鼠抑郁樣行為的作用及機制
發(fā)布時間：2018-10-23 20:26
【摘要】：第一部分硫化氫對慢性不可預(yù)知性應(yīng)激誘導的大鼠抑郁樣行為的作用 目的：硫化氫(hydrogen sulfide,H2S)是哺乳動物體內(nèi)的一種內(nèi)源性氣體信號分子,在中樞神經(jīng)系統(tǒng)具有廣泛的生物學效應(yīng)。研究報道,阿爾茨海默病、帕金森病、腦卒中等疾病的產(chǎn)生皆與腦內(nèi)H2S水平失調(diào)有關(guān)。然而,腦內(nèi)H2S的含量變化是否參與抑郁癥的發(fā)病過程,外源性調(diào)控腦內(nèi)H2S含量是否對抑郁癥的治療具有促進作用,目前鮮有報道。因此,本實驗將探討抑郁癥大鼠腦內(nèi)H2S水平的變化,并研究外源性補充H2S對大鼠抑郁樣行為的作用。 方法：應(yīng)用性不可預(yù)知性溫和應(yīng)激(chronic unpredictable mild stress, CUMS)大鼠模型進行研究；采用氣相色譜-質(zhì)譜聯(lián)用儀(gas chromatograph-mass spectrometer, GC-MS)檢測CUMS大鼠海馬腦區(qū)內(nèi)源性H2S水平的改變；采用糖水偏好實驗(sucrose preference test, SPT)、新奇環(huán)境攝食抑制實驗(novelty-suppressed feeding test, NSFT).強迫游泳實驗(forced swimming test. FST)觀察大鼠抑郁樣行為；采用高爾基染色觀察H2S對CUMS大鼠海馬神經(jīng)元樹突棘密度的影響；采用酶聯(lián)免疫吸附測定(enzyme linked immunosorbent assay, ELISA)試劑盒來檢測CUMS大鼠血清皮質(zhì)酮水平。 結(jié)果：CUMS大鼠海馬腦區(qū)H2S含量顯著降低。腹腔注射硫氫化鈉(NaHS,11.2mg/kg)、吸入H2S氣體(3小時,120ppm)兩種給藥方式,均可升高CUMS大鼠海馬腦區(qū)H2S水平。腹腔注射硫氫化鈉(sodium hydrosulfide, NaHS)顯著降低正常大鼠急性FST的不動時間,該作用在30分鐘內(nèi)即可快速起效,且藥效能夠維持一周,其效果優(yōu)于NMDA受體拮抗藥MK-801和氟西汀。腹腔注射NaHS對CUMS誘導的大鼠抑郁樣行為也有改善作用,即增加CUMS大鼠SPT的糖水偏好率、縮短NSFT的攝食潛伏期、降低FST的不動時間。此外,H2S可減輕CUMS所致的大鼠海馬神經(jīng)元樹突棘密度的降低,但對大鼠血清皮質(zhì)酮水平無明顯影響。 結(jié)論：慢性應(yīng)激降低大鼠海馬腦區(qū)H2S水平,而外源性補充H2S對急、慢性應(yīng)激導致的大鼠抑郁樣行為均有改善作用,提示調(diào)控海馬腦區(qū)H2S含量可能是治療抑郁癥的新策略。 第二部分硫化氫拮抗慢性不可預(yù)知性應(yīng)激誘導的大鼠抑郁樣行為的機制 目的：近年來,氯胺酮的抗抑郁作用的發(fā)現(xiàn),推動了快速起效抗抑郁藥物的研究并取得了突破性進展,而對快速起效抗抑郁藥物機制的探索也在不斷深入。哺乳動物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信號及其下游通路的激活,對藥物產(chǎn)生快速的抗抑郁作用具有重要意義。氣體信號分子H2S所產(chǎn)生的快速抗抑郁樣行為的作用機制是否與mTOR信號的激活有關(guān)?本部分實驗將進行研究。 方法：應(yīng)用蛋白質(zhì)印跡法(Western blot)檢測H2S對CUMS大鼠海馬腦區(qū)mTOR信號下游的P70S6k的調(diào)控作用,通過藥理學阻斷和分子生物學技術(shù),并結(jié)合行為學檢測,觀察H2S的抗抑郁作用并探討其作用機制。 結(jié)果：Western blot實驗發(fā)現(xiàn)H2S可迅速激活海馬腦區(qū)哺乳動物雷帕霉素靶蛋白復(fù)合物1(mammalian target of rapamycin complex1, mTORC1)信號下游的核糖體蛋白S6激酶70kD (ribosomal protein S6kinases70kD, P70S6k)。而通過mTORC1特異性阻斷劑雷帕霉素(rapamycin)或慢病毒載體構(gòu)建mTOR調(diào)節(jié)相關(guān)蛋白(regulatory associated protein of mTOR, Raptor)的shRNA阻斷mTORC1,并結(jié)合行為學檢測,發(fā)現(xiàn)H2S的抗抑郁作用與mTORC1的激活有關(guān)。此外,H2S促進突觸后密度95(postsynaptic density95, PSD95)、突觸素(synaptophysin)的合成,并通過上調(diào)酪氨酸激酶受體B (tyrosine kinase receptor B, TrkB)表達,促進AMPA受體GluR1亞基總蛋白及磷酸化蛋白水平。 結(jié)論：H2S通過激活mTORC1信號通路,促進突觸蛋白合成,并上調(diào)TrkB受體,進一步促進AMPA受體GluRl亞基總蛋白及磷酸化蛋白水平,從而發(fā)揮抗抑郁作用。
[Abstract]:Effect of the first part of hydrogen sulfide on depression-like behavior in rats induced by chronic unpredictable stress Objective: Hydrogen sulfide (H2S) is a kind of endogenous gas signal molecule in mammals, which has extensive biology in the central nervous system. Effect: It is reported that the generation of Alzheimer's disease, Parkinson's disease, stroke and other diseases is associated with the level of H2S in the brain. However, whether the content of H2S in the brain is involved in the pathogenesis of depression, the exogenous regulation of the content of H2S in the brain can promote the treatment of depression, Therefore, this study will explore the changes of the levels of H2S in the brain of depression rats and study the effects of exogenous supplemental H2S on depression-like behavior in rats. Methods: The model of the rat model was studied by using gas chromatography-mass spectrometry (GC-MS). Gas chromatography-mass spectrometry (GC-MS) was used to detect the changes of endogenous H2S levels in hippocampus of CUMS rats. SPT), novel environment feeding inhibition experiment (SPT), NSFT). Forced swimming test (for swimmin) g test. FST) observation of depression-like behavior in rats; observation of the effect of H2S on the dendritic spines density of hippocampal neurons in CUMS rats by Golgi staining; ELISA kits were used to detect the serum of CUMS rats. Corticosterone levels. Results: brain region H of hippocampus in CUMS rats The content of 2S was significantly reduced, and both modes of administration of H2S gas (NaHS, 11.2mg/ kg) and H2S gas (3 hours, 120ppm) could increase the sea level of CUMS rats. The levels of H2S in the brain of horse brain were significantly lower than that in normal rats by intraperitoneal injection of sulfocyanide (NaHS). The effect could quickly take effect within 30 minutes, and the effect could be maintained for one week. The effect was better than that of NMDA receptor antagonist. The effect of NaHS on the depression-like behavior of the rats induced by CUMS was also improved, that is to increase the rate of appetite of the SPTs in the CUMS rats, shorten the feeding latency of the NSFT, and decrease the feeding latency of the NSFT rats. In addition, H2S can reduce the decrease of dendritic spines density in rat hippocampal neurons induced by CUMS, Conclusion: Chronic stress reduces the level of H2S in hippocampus of rats, while exogenous supplemental H2S has an improved effect on depression-like behavior in rats with acute and chronic stress. It is suggested that the content of H2S in brain region of hippocampus can be regulated and controlled. can be a new strategy for treating depression, and the second part of hydrogen sulfide antagonizes chronic unpredictable The mechanism of stress-induced depression-like behavior in rats: the discovery of anti-depressant effects of ketamine in recent years has contributed to rapid onset of action The study of anti-depressant drugs has made breakthrough progress, and fast The study of the mechanism of anti-depressant drugs has also been deepening. Activation of the mTOR signal and its downstream pathway in mammalian Repamycin target protein (mTOR) signal and its downstream pathway The rapid antidepressant action of the substance is of great significance. The rapid antidepressant action produced by the molecular H2S of the gas signal Whether or not the mechanism works with mTOR The effects of H2S on the P70S6k downstream of the mTOR signal in the hippocampus of CUMS rats were detected by Western blot. Results: Western blot showed that H2S can rapidly activate the ribosomal protein S6 kinase 70kD (ribosomal prote) downstream of the MTORC1 signal. in S6kinases70kD, P70S6k). It was found that the antidepressant effect of H2S is related to the activation of mTORC1. In addition, H2S promotes the synthesis of postsynaptic density 95 (PSD95) and synaptosin and promotes A by up-regulating the expression of tyrosine kinase receptor B (TrkB). Conclusion: H2S can promote the synthesis of synaptosin by activating mTORC1 signaling pathway and upregulate the TrkB receptor to further promote AMPA receptor Gl.
【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R965
[Abstract]:Effect of the first part of hydrogen sulfide on depression-like behavior in rats induced by chronic unpredictable stress Objective: Hydrogen sulfide (H2S) is a kind of endogenous gas signal molecule in mammals, which has extensive biology in the central nervous system. Effect: It is reported that the generation of Alzheimer's disease, Parkinson's disease, stroke and other diseases is associated with the level of H2S in the brain. However, whether the content of H2S in the brain is involved in the pathogenesis of depression, the exogenous regulation of the content of H2S in the brain can promote the treatment of depression, Therefore, this study will explore the changes of the levels of H2S in the brain of depression rats and study the effects of exogenous supplemental H2S on depression-like behavior in rats. Methods: The model of the rat model was studied by using gas chromatography-mass spectrometry (GC-MS). Gas chromatography-mass spectrometry (GC-MS) was used to detect the changes of endogenous H2S levels in hippocampus of CUMS rats. SPT), novel environment feeding inhibition experiment (SPT), NSFT). Forced swimming test (for swimmin) g test. FST) observation of depression-like behavior in rats; observation of the effect of H2S on the dendritic spines density of hippocampal neurons in CUMS rats by Golgi staining; ELISA kits were used to detect the serum of CUMS rats. Corticosterone levels. Results: brain region H of hippocampus in CUMS rats The content of 2S was significantly reduced, and both modes of administration of H2S gas (NaHS, 11.2mg/ kg) and H2S gas (3 hours, 120ppm) could increase the sea level of CUMS rats. The levels of H2S in the brain of horse brain were significantly lower than that in normal rats by intraperitoneal injection of sulfocyanide (NaHS). The effect could quickly take effect within 30 minutes, and the effect could be maintained for one week. The effect was better than that of NMDA receptor antagonist. The effect of NaHS on the depression-like behavior of the rats induced by CUMS was also improved, that is to increase the rate of appetite of the SPTs in the CUMS rats, shorten the feeding latency of the NSFT, and decrease the feeding latency of the NSFT rats. In addition, H2S can reduce the decrease of dendritic spines density in rat hippocampal neurons induced by CUMS, Conclusion: Chronic stress reduces the level of H2S in hippocampus of rats, while exogenous supplemental H2S has an improved effect on depression-like behavior in rats with acute and chronic stress. It is suggested that the content of H2S in brain region of hippocampus can be regulated and controlled. can be a new strategy for treating depression, and the second part of hydrogen sulfide antagonizes chronic unpredictable The mechanism of stress-induced depression-like behavior in rats: the discovery of anti-depressant effects of ketamine in recent years has contributed to rapid onset of action The study of anti-depressant drugs has made breakthrough progress, and fast The study of the mechanism of anti-depressant drugs has also been deepening. Activation of the mTOR signal and its downstream pathway in mammalian Repamycin target protein (mTOR) signal and its downstream pathway The rapid antidepressant action of the substance is of great significance. The rapid antidepressant action produced by the molecular H2S of the gas signal Whether or not the mechanism works with mTOR The effects of H2S on the P70S6k downstream of the mTOR signal in the hippocampus of CUMS rats were detected by Western blot. Results: Western blot showed that H2S can rapidly activate the ribosomal protein S6 kinase 70kD (ribosomal prote) downstream of the MTORC1 signal. in S6kinases70kD, P70S6k). It was found that the antidepressant effect of H2S is related to the activation of mTORC1. In addition, H2S promotes the synthesis of postsynaptic density 95 (PSD95) and synaptosin and promotes A by up-regulating the expression of tyrosine kinase receptor B (TrkB). Conclusion: H2S can promote the synthesis of synaptosin by activating mTORC1 signaling pathway and upregulate the TrkB receptor to further promote AMPA receptor Gl.
【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R965
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