本文關(guān)鍵詞：雷帕霉素對大鼠嬰兒期遺忘作用的研究　出處：《南方醫(yī)科大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 嬰兒期遺忘 雷帕霉素 齒狀回 神經(jīng)發(fā)生

【摘要】：研究背景早期生活經(jīng)歷對成年后人格和心理社會功能的養(yǎng)成起著非常重要的作用,然而成人仍很少能回憶起幼年時期發(fā)生的事件。嬰兒期遺忘(infantile amnesia)是指成人對特定年齡前發(fā)生的生活事件無法進(jìn)行回憶的現(xiàn)象,部分研究則定義為與成年期相比,嬰幼兒時期/兒童期對記憶自發(fā)性遺忘的加速現(xiàn)象(在兒童期為童年期遺忘)。目前人類嬰兒期遺忘的邊界年齡尚未明確,人類方面的研究主要集中在神經(jīng)心理學(xué)方面。1962年Campbell利用行為學(xué)實驗(主動回避反應(yīng)訓(xùn)練)驗證了嬰兒期遺忘現(xiàn)象同樣存在于大鼠身上,此后,使用不同種屬的實驗動物(如靈長類)和行為學(xué)訓(xùn)練方法(如被動回避反應(yīng)訓(xùn)練、條件性恐懼訓(xùn)練)的實驗均驗證了嬰兒期遺忘現(xiàn)象的存在,為更好地研究其發(fā)生機(jī)制奠定了基礎(chǔ)。目前尚未明確嬰兒期遺忘發(fā)生的神經(jīng)生物學(xué)機(jī)制,但明確的是其發(fā)生機(jī)制并非單一,可能涉及眾多與記憶相關(guān)的腦區(qū)、神經(jīng)遞質(zhì)系統(tǒng)和神經(jīng)內(nèi)分泌系統(tǒng)等。海馬、內(nèi)側(cè)前額葉皮質(zhì)均是晚成熟結(jié)構(gòu),N-甲基-D-天冬氨酸受體(N-methyl-D-aspartic acid, NMD A)、γ-氨基丁酸(gamma-aminobutyric acid, GABA)等受體存在亞單位發(fā)育的時空差異等,均可能影響嬰幼兒時期的學(xué)習(xí)記憶功能。海馬屬于邊緣系統(tǒng)結(jié)構(gòu),也是晚成熟結(jié)構(gòu),出生后海馬仍需經(jīng)歷神經(jīng)發(fā)生、突觸發(fā)生、神經(jīng)遞質(zhì)受體發(fā)育、髓鞘形成等解剖結(jié)構(gòu)和生理功能的改變。情景式的學(xué)習(xí)記憶如穿梭箱被動回避反應(yīng)、條件性恐懼訓(xùn)練均是海馬依賴性的學(xué)習(xí)記憶過程,均需完整的海馬功能的參與。Lavenex P等通過測試不同年齡兒童的空間學(xué)習(xí)記憶,并對比靈長類動物隨年齡增長而改變的海馬結(jié)構(gòu),發(fā)現(xiàn)嬰兒期遺忘時間與海馬結(jié)構(gòu)功能的發(fā)育差異時間基本一致,提出兩者可能存在因果關(guān)系。Josselyn SA等也認(rèn)為海馬的發(fā)育差異在嬰兒期遺忘中起著重要作用,猜測嬰幼兒時期海馬高水平的神經(jīng)發(fā)生可能與嬰兒期的加速遺忘現(xiàn)象相關(guān)。海馬神經(jīng)發(fā)生與學(xué)習(xí)記憶相關(guān),在特定情況下,成年海馬神經(jīng)發(fā)生可促進(jìn)依賴海馬的新記憶的形成和保留,但對于已形成的舊記憶,過多新生神經(jīng)元對已有神經(jīng)環(huán)路的持續(xù)整合卻可能加速原有記憶的遺忘,新生的突觸連接也可能對原有環(huán)路造成干擾。海馬齒狀回是大腦神經(jīng)發(fā)生的主要區(qū)域之一,齒狀回神經(jīng)發(fā)生貫穿于生命體的整個生命過程并隨年齡增長而水平逐漸降低,嬰幼兒時期神經(jīng)發(fā)生水平較高而成年后水平降低。嬰幼兒期高水平的神經(jīng)發(fā)生與記憶保留呈負(fù)相關(guān)關(guān)系在Akers等的研究中同樣得到證實：使用17日齡和60日齡小鼠進(jìn)行條件性恐懼記憶訓(xùn)練,免疫熒光染色檢測神經(jīng)發(fā)生水平,結(jié)果顯示17日齡小鼠比60日齡小鼠有更高水平的海馬神經(jīng)發(fā)生,且17日齡小鼠對恐懼記憶的遺忘速度比60日齡快；通過基因和藥理學(xué)途徑減少17日齡小鼠的神經(jīng)發(fā)生可增加其記憶保留；而通過自發(fā)輪轉(zhuǎn)運(yùn)動增加60日齡小鼠的神經(jīng)發(fā)生卻加速其記憶遺忘,提示嬰幼兒期過高水平的神經(jīng)發(fā)生可能是嬰兒期遺忘的發(fā)生機(jī)制之一。哺乳動物雷帕霉素靶蛋白(mammalian target of rapamycin, mTOR)是由2549個氨基酸組成的進(jìn)化上相對保守的絲氨酸/蘇氨酸激酶。mTOR信號通路可接受激素、生長因子、營養(yǎng)因子和能量等多種信號輸入,通過PI3K/Akt/mTOR或AMPK等途徑來發(fā)揮控制基因轉(zhuǎn)錄和蛋白質(zhì)翻譯起始、調(diào)控細(xì)胞周期、調(diào)節(jié)凋亡和影響代謝等多重作用。雷帕霉素是mTOR的抑制劑,可通過阻止mTOR通路下游效應(yīng)器P70S6K和4EBPs的磷酸化,抑制相關(guān)蛋白的轉(zhuǎn)錄和翻譯,起著影響免疫、調(diào)控細(xì)胞增殖、調(diào)節(jié)凋亡與自噬等作用。據(jù)國外的相關(guān)研究,雷帕霉素腹腔注射可影響幼鼠海馬齒狀回新生神經(jīng)元的增殖。雷帕霉素亦可影響突觸可塑性,影響學(xué)習(xí)記憶等相關(guān)過程。動物實驗表明,局部腦區(qū)內(nèi)(如海馬、杏仁核、皮層)注射或腹腔注射雷帕霉素可損害依賴海馬的空間記憶、依賴前額葉皮層或杏仁核的條件恐懼記憶、依賴味覺皮層的條件味覺厭惡記憶等學(xué)習(xí)記憶過程。而另一方面,雷帕霉素長期低劑量口服則能增強(qiáng)成年小鼠的空間學(xué)習(xí)記憶和老年鼠被動回避反應(yīng)訓(xùn)練的記憶保留,改善阿爾茨海默病模型小鼠的記憶損害,增加老年鼠的平均壽命。綜合以上研究,提示雷帕霉素對學(xué)習(xí)記憶的影響可能與身體狀態(tài)、年齡、用藥途徑等有關(guān)。目前甚少研究關(guān)注于探討雷帕霉素對幼齡大鼠學(xué)習(xí)記憶的影響。由于雷帕霉素可影響幼齡大鼠海馬齒狀回新生神經(jīng)元的增殖,而海馬神經(jīng)發(fā)生又與記憶保留相關(guān),推測雷帕霉素或通過影響神經(jīng)發(fā)生作用于嬰兒期遺忘。因此,本實驗通過研究雷帕霉素腹腔注射對幼齡大鼠被動回避反應(yīng)訓(xùn)練即學(xué)習(xí)記憶的影響,同時探討雷帕霉素腹腔注射對幼齡大鼠海馬細(xì)胞增殖、神經(jīng)發(fā)生和細(xì)胞凋亡的作用,初步探討雷帕霉素影響嬰兒期遺忘的可能機(jī)制。具體研究包括以下兩部分：第一部分：雷帕霉素對幼齡大鼠被動回避反應(yīng)訓(xùn)練的影響目的穿梭箱被動回避反應(yīng)訓(xùn)練17日齡(post-natal day 17, P17)和60日齡(post-natal day 60, P60)大鼠,驗證嬰兒期遺忘現(xiàn)象。研究雷帕霉素腹腔注射對17日齡大鼠被動回避反應(yīng)訓(xùn)練記憶保留的影響。方法(1)17日齡和60日齡SD雄性大鼠各42只,各年齡組根據(jù)被動回避反應(yīng)訓(xùn)練時是否給予電擊又分為電擊組與非電擊組,測試各組大鼠被動回避反應(yīng)訓(xùn)練后O d(即訓(xùn)練剛結(jié)束后)、2d和7d的記憶保留。(2)P17 SD雄性大鼠84只,分為溶媒組、雷帕霉素20mg/kg(Rap20組)和雷帕霉素40mg/kg組(Rap40組),各組分別在被動回避反應(yīng)訓(xùn)練后腹腔注射等體積的溶媒或雷帕霉素溶液。各組根據(jù)被動回避反應(yīng)訓(xùn)練時是否給予電擊又分電擊組與非電擊組,測試訓(xùn)練后2 d(P19)和7 d(P24)的記憶保留。結(jié)果(1)17和60日齡大鼠被動回避反應(yīng)訓(xùn)練結(jié)果的差異研究適應(yīng)階段反應(yīng)潛時P17和P60大鼠比較差異無統(tǒng)計學(xué)意義(P0.05)。測試時電擊組2×3析因分析提示不同年齡組差異有統(tǒng)計學(xué)意義(P0.05),測試時不同時間點(diǎn)差異有統(tǒng)計學(xué)意義(P0.05)。電擊組訓(xùn)練后0d(即訓(xùn)練結(jié)束后)P17和P60大鼠反應(yīng)潛時比較差異無統(tǒng)計學(xué)意義(P0.05),訓(xùn)練后2d和7d兩年齡組反應(yīng)潛時比較差異均有統(tǒng)計學(xué)意義(P0.05)。P17大鼠訓(xùn)練后2d記憶保留有所減弱,與訓(xùn)練后0d反應(yīng)潛時比較差異有統(tǒng)計學(xué)意義(P0.05),到訓(xùn)練后7d差異更明顯,反應(yīng)潛時下降至接近電擊訓(xùn)練前水平。P60大鼠在訓(xùn)練后2d和7d仍保留穩(wěn)定的記憶水平,與訓(xùn)練后Od反應(yīng)潛時比較差異均無統(tǒng)計學(xué)意義(P0.05)。非電擊組P17和P60大鼠均在測試各時間點(diǎn)(訓(xùn)練后0d、2d和7 d)顯示出與適應(yīng)階段無明顯差異的反應(yīng)潛時(P0.05)。(2)雷帕霉素對P17大鼠被動回避反應(yīng)訓(xùn)練的影響適應(yīng)階段反應(yīng)潛時溶媒組、Rap20組和Rap40組比較差異無統(tǒng)計學(xué)意義(P0.05)。測試時電擊組3×2析因分析提示不同劑量組差異有統(tǒng)計學(xué)意義(P0.05),測試時不同時間點(diǎn)差異有統(tǒng)計學(xué)意義(P0.05)。電擊組訓(xùn)練后2d三組反應(yīng)潛時比較差異無統(tǒng)計學(xué)意義(P0.05)；訓(xùn)練后7d反應(yīng)潛時Rap20組、Rap40組與溶媒組比較差異均有統(tǒng)計學(xué)意義(P0.05),即雷帕霉素用藥組增加了P17大鼠被動回避反應(yīng)訓(xùn)練的反應(yīng)潛時,而溶媒組在訓(xùn)練后7d反應(yīng)潛時已下降至接近電擊訓(xùn)練前水平。Rap20組和Rap40組在訓(xùn)練后2d和7d反應(yīng)潛時比較差異均無統(tǒng)計學(xué)意義(P0.05)。非電擊組各組各時間點(diǎn)反應(yīng)潛時比較差異均無統(tǒng)計學(xué)意義(P0.05)。結(jié)論17日齡大鼠存在快速遺忘,顯示了嬰兒期遺忘現(xiàn)象。雷帕霉素腹腔注射可增加P17大鼠對電擊訓(xùn)練的記憶保留,減緩嬰兒期遺忘現(xiàn)象。第二部分：雷帕霉素對幼齡大鼠海馬齒狀回細(xì)胞增殖、神經(jīng)發(fā)生和細(xì)胞凋亡的影響目的探討雷帕霉素腹腔注射對17日齡大鼠海馬齒狀回細(xì)胞增殖、神經(jīng)發(fā)生和細(xì)胞凋亡的影響,初步探討雷帕霉素影響幼齡大鼠記憶保留的可能機(jī)制。方法(1)P17 SD雄性大鼠24只,分為正常組、溶媒組、雷帕霉素20mg/kg組(Rap20組)和40mg/kg組(Rap40組),于17日齡同時間點(diǎn)分別腹腔注射等體積的溶媒或雷帕霉素溶液(正常組不予處理),注射44 h后所有動物腹腔注射BrdU溶液標(biāo)記增殖細(xì)胞,劑量為100mg/kg,于BrdU注射4h后(即P19)每組6只大鼠灌注取腦,行海馬齒狀回BrdU免疫熒光單標(biāo)染色。(2)P17 SD雄性大鼠48只,分為正常組、溶媒組、Rap20組和Rap40組,于17日齡同時間點(diǎn)分別腹腔注射等體積的溶媒或雷帕霉素溶液(正常組不予處理),注射12h后所有動物腹腔注射BrdU溶液標(biāo)記增殖細(xì)胞,劑量為50mg/kg,每6h注射一次,共5次,于BrdU末次注射12h和132 h后(即P19和P24),每組各時間點(diǎn)取6只大鼠灌注取腦,行海馬齒狀回BrdU/DCX免疫熒光雙標(biāo)染色。(3)P17 SD雄性大鼠24只,分為正常組、溶媒組、Rap20組和Rap40組,于17日齡同時間點(diǎn)分別腹腔注射等體積的溶媒或雷帕霉素溶液(正常組不予處理),于注射藥物48 h后(即P19)每組6只大鼠灌注取腦,石蠟切片行海馬齒狀回caspase3免疫組化染色。結(jié)果(1)溶媒組與Rap20組、Rap40組BrdU陽性細(xì)胞計數(shù)比較差異均有統(tǒng)計學(xué)意義(P0.05),雷帕霉素能減少海馬齒狀回的細(xì)胞增殖。BrdU陽性細(xì)胞計數(shù)正常組與溶媒組比較差異無統(tǒng)計學(xué)意義(P0.05), Rap20組與Rap40組比較差異有統(tǒng)計學(xué)意義(P0.05), Rap40組比Rap20組BrdU陽性細(xì)胞計數(shù)少。(2)溶媒組與Rap20組、Rap40組BrdU、BrdU/DCX陽性細(xì)胞計數(shù)在各時間點(diǎn)(P19和P24)比較差異均有統(tǒng)計學(xué)意義(P0.05),雷帕霉素能減少海馬齒狀回的神經(jīng)發(fā)生。BrdU、BrdU/DCX陽性細(xì)胞計數(shù)正常組與溶媒組各時間點(diǎn)比較差異均無統(tǒng)計學(xué)意義5), Rap20組與Rap40組各時間點(diǎn)比較差異均有統(tǒng)計學(xué)意義(P0.05),增加雷帕霉素的劑量陽性細(xì)胞計數(shù)更少。P24與P19相比,各組BrdU、BrdU/DCX陽性細(xì)胞計數(shù)均有不同程度減少。(3)溶媒組與Rap20組、Rap40組caspase3陽性細(xì)胞計數(shù)比較差異均有統(tǒng)計學(xué)意義(P0.05),雷帕霉素能增加海馬齒狀回的細(xì)胞凋亡。Caspase3陽性細(xì)胞計數(shù)正常組與溶媒組比較差異無統(tǒng)計學(xué)意義(P0.05), Rap20組與Rap40組比較差異有統(tǒng)計學(xué)意義(P0.05),Rap40組caspase3陽性細(xì)胞計數(shù)比Rap20組多。結(jié)論雷帕霉素能減少P17幼齡大鼠海馬齒狀回的細(xì)胞增殖和神經(jīng)發(fā)生,增加海馬齒狀回的細(xì)胞凋亡。
[Abstract]:On the background of early life experience to develop personality and psychosocial functioning in adulthood plays a very important role, but adults are still rarely recalled childhood events. Infantile amnesia (infantile amnesia) refers to the adult age specific life before the event can not recall phenomenon, part of the research is defined as compared with adults, children of spontaneous forgetting to accelerate the phenomenon of memory in infancy (/ forgotten childhood in childhood). The human infants forgetting the boundary age is not yet clear, the human aspects of the research mainly concentrated in the aspects of neuropsychological.1962 Campbell using behavioral tests (active avoidance reaction) verification the infantile amnesia phenomenon also exists in the rat body, then use different species of animal (such as primates) and training methods (such as passive avoidance behavior Response training, conditioned fear training) experiments have verified the existence of the phenomenon of infantile amnesia, laid the foundation for better research on its mechanism is not yet clear. Infantile amnesia neurobiological mechanisms occur, but its mechanism is not clear is single, may involve a number of memory related brain regions, neurotransmitter system and neuroendocrine system. The hippocampus and medial prefrontal cortex are late maturity structure, N- methyl -D- aspartate receptor (N-methyl-D-aspartic acid, NMD A), gamma aminobutyric acid (gamma-aminobutyric, acid, GABA) are the development of subunit space-time difference etc. receptor, can affect the function of learning and memory in infancy. The hippocampus belongs to the limbic system, and late maturity structure, still need to experience after the birth of hippocampal neurogenesis, synaptogenesis, neurotransmitter receptor development, myelin formation anatomy The structure and physiological function. The change of learning and memory of scenes such as passive avoidance test, conditioned fear training is the process of learning and memory in hippocampus dependent, are required to complete the.Lavenex P in the hippocampal function through the test of learning and memory of children of different ages in space, hippocampus and contrast of primate animal with age change, found time differential development of infantile amnesia and hippocampal structure and function of the basic agreement, put forward the possible causal relationship between the.Josselyn SA also believes that the developmental differences in infant hippocampus plays an important role in the forgetfulness, speculation during high levels of hippocampal neurogenesis may be associated with infant hippocampus accelerated forgetting phenomenon. Neurogenesis is associated with learning and memory, in certain circumstances, the formation of new adult hippocampal neurogenesis in the hippocampus dependent memory can promote and retain, But for the formation of the old memory, too many new neurons continued integration of existing neural circuits may accelerate the original forgotten memory, connect new synapses can also cause interference to the original loop. The dentate gyrus is one of the main areas of the brain neurogenesis, neurogenesis in the dentate gyrus throughout the whole process of life and gradually decreased with age, infants higher levels of adult neurogenesis and neural levels. High levels of infant and memory retention was negatively related in the research of Akers in the same confirmed: the use of 17 day old and 60 day old mice were conditioned fear memory training, immunofluorescence staining neurogenesis level, the results showed that 17 day old mice had higher levels of hippocampal neurogenesis than 60 day old mice, and 17 day old mice on fear of forgetting At the age of 60 days faster than fast; by genetic and pharmacological approaches to reduce 17 day old mice neurogenesis can increase the memory retention; and through the self rotation rotary motion increase of 60 day old mice have accelerated the neural amnesia, suggesting that infants high levels of neurogenesis may be the pathogenesis of infantile amnesia. The mammalian target of rapamycin (mammalian target of rapamycin, mTOR) is a relatively conservative consisting of 2549 amino acids on the evolution of serine / threonine kinase.MTOR signaling pathway can receive hormone, growth factor, nutrition factor and energy and other inputs, through PI3K/Akt/mTOR or AMPK and other ways to exert control of gene transcription and protein translation initiation. The regulation of cell cycle regulation, apoptosis and metabolism of multiple roles. Rapamycin is an inhibitor of mTOR, by blocking mTOR pathway downstream. Is P70S6K and the phosphorylation of 4EBPs, inhibition of transcription and translation related protein, plays influence immune, regulation of cell proliferation, apoptosis and autophagy. According to the related research abroad, rapamycin intraperitoneal injection can affect neurogenesis in the dentate gyrus of rats. The proliferation effect of synaptic plasticity may influence the learning and memory of rapamycin. Other related process. Animal experiments show that local brain regions (such as the hippocampus, amygdala, cortex) spatial memory injection or intraperitoneal injection of rapamycin can damage the hippocampus dependent fear conditioning, dependent prefrontal cortex or amygdala dependent, gustatory cortex conditioned taste aversion memory learning and memory process. On the other hand, long-term low rapamycin oral dose of reinforcement learning can passive avoidance response training and memory of aged rats memory retention in adult mice, improve Alzheimer's disease model The memory impairment of mice, increase the life expectancy of aged rats. Based on the above study, suggesting that effects of rapamycin on learning and memory and physical condition, age, route of administration and so on. There are few studies focus on the effects of rapamycin on learning and memory in young rats. The effects of rapamycin in young rats neurogenesis in the dentate gyrus the proliferation and hippocampal neurogenesis and memory retention, presumably through the influence of rapamycin or neurogenesis in infantile amnesia. Therefore, through the study of rapamycin intraperitoneal injection on rat passive avoidance response training effects of learning and memory, and discusses on the proliferation of hippocampal cells in young rats by intraperitoneal injection of rapamycin, neurogenesis and the role of apoptosis, to investigate the possible mechanisms of the effects of rapamycin in infancy forgotten. Specific studies include in The two part: the first part: the influence of rapamycin on young rats of passive avoidance response training to passive avoidance test in training at the age of 17 days (post-natal day 17, P17) and 60 days (post-natal day 60, P60) rats, verification of infantile amnesia phenomenon. Research on ray rapamycin intraperitoneal injection of 17 day old rats passive avoidance reaction training memory retention. Methods (1) 17 day old and 60 day old male SD rats 42 rats in each age group, the passive avoidance according to whether or not to give electric shocks and divided into shock group and shock group response training, all rats were tested after training passive avoidance response O D (i.e. just after the end of training), 2D and 7d (2) P17 memory retention. 84 male SD rats, divided into solvent group, rapamycin 20mg/kg (Rap20 group) and rapamycin group 40mg/kg (Rap40 group), each group were in passive avoidance of solvent was injected into the reaction volume after training Or rapamycin solution. Each group based on passive avoidance reaction to shocks and whether the training is divided into non shock group and shock group, test after training 2 D (P19) and 7 d (P24) memory retention. Results (1) 17 and the difference between the 60 day old rats of passive avoidance response training results Research on the adaptive phase of the reaction potential when P17 and P60 rats showed no significant difference (P0.05). The test shock group 2 * 3 factorial analysis showed that the differences between different age groups was statistically significant (P0.05), when tested at different time points were statistically significant (P0.05). 0d shock group after training (i.e. after training) and P60 P17 rat response latency had no significant difference (P0.05), 2D and 7d after training two age group formation. There were significant differences (P0.05) in.P17 rats after training 2D memory retention weakened, and potential 0d response after the training when the difference was statistically significant (P0.05), The difference is more obvious after 7d training, the reaction latency dropped to near the level before the shock training.P60 rats after training in 2D and 7d remained stable memory level, and potential Od reaction after training showed no significant difference (P0.05). The non shock group P17 and P60 rats were tested in different time point (after training 0d 2D and 7 d) reaction shows no significant difference between the latent phase of adaptation (P0.05). (2) the effect of rapamycin on P17 rat passive avoidance reaction influence training adaptation stage of the reaction potential when the solvent group, there was no significant difference between Rap20 group and Rap40 group (P0.05) test. When the electric group of 3 * 2 factorial analysis showed that different dose groups were statistically significant (P0.05), when tested at different time points were statistically significant (P0.05). The shock training group 2D three group reaction latency had no significant difference (P0.05); potential 7d after training in Rap20 group, Rap40 Group and solvent group were statistically significant (P0.05), which increases the reaction potential rapamycin treatment group training passive avoidance response in P17 rats, and solvent group in the latent 7d reaction after training has dropped to near the level before the shock training group.Rap20 and Rap40 group after training 2D and 7d response latency there were no significant differences (P0.05). The non shock group at each time point response latency showed no significant difference (P0.05). The conclusion of the 17 day old rats are quickly forgotten, shows the phenomenon of infantile amnesia. Rapamycin intraperitoneal injection can increase the memory of the shock training P17 rats retained, slow down baby during the period of forgetting phenomenon. The second part: rapamycin on young rat dentate gyrus cell proliferation, neurogenesis and apoptosis effect study of rapamycin intraperitoneal injection of 17 day old rat dentate gyrus cell proliferation, Effect of neurogenesis and apoptosis, to investigate the possible mechanisms of memory in young rats. Methods the retention effect of rapamycin (1) P17 24 male SD rats were divided into normal group, solvent group, rapamycin group 20mg/kg (Rap20 group) and 40mg/kg group (Rap40 group), at the age of 17 at the same time. Intraperitoneal injection of equal volume of solvent or rapamycin solution (the normal group without treatment), 44 h after injection of all animal injected BrdU labeled cells, the dose of 100mg/kg, BrdU after injection of 4H (P19) 6 rats in each group underwent brain perfusion, dentate gyrus BrdU immunofluorescence single stained. (2) P17 48 male SD rats were divided into normal group, vehicle group, Rap20 group and Rap40 group, at the age of 17 at the same time were injected equal volume of solvent or rapamycin solution (the normal group without treatment), 12h after injection of all animal injected BrdU labeling. Cell dose is 50mg/kg, each 6h injection time, a total of 5 times, in the end of BrdU and 132 h after injection of 12h (P19 and P24), each group of each time point, 6 rats were perfused and brains, for dentate gyrus BrdU/DCX immunofluorescence staining. (3) P17 SD male 24 rats were divided into normal group, vehicle group, Rap20 group and Rap40 group, at the age of 17 at the same time were injected equal volume of solvent or rapamycin solution (the normal group without treatment), 48 h after drug injection (P19) with 6 rats in each group perfused brain, paraffin sections. The dentate gyrus Caspase3 immunohistochemical staining. Results (1) the solvent group and Rap20 group, Rap40 group, BrdU positive cells were statistically significant (P0.05), rapamycin can reduce dentate cell proliferation of.BrdU positive cells in normal group and solvent group was not statistically significance (P0.05). Rap20 group and Rap40 group difference There was a significant correlation (P0.05), Rap40 group than in the Rap20 group BrdU positive cells. (2) the solvent group and Rap20 group, Rap40 group, BrdU, BrdU/DCX positive cells at different time points (P19 and P24) showed no significant differences (P0.05), rapamycin can reduce dentate gyrus neurogenesis.BrdU, the number of BrdU/DCX positive cells in normal group and solvent group at each time point, there were no significant differences in 5), Rap20 group and Rap40 group at different time points were statistically significant (P0.05), increase the dose of rapamycin positive cells count less.P24 compared with P19, BrdU group, BrdU/DCX positive cell counts were not

【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R965

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陳同度,張昌穎;素食大鼠的貧血現(xiàn)象[J];營養(yǎng)學(xué)報;1957年04期

2 陳偉強(qiáng);趙善廣;;自制注射用大鼠固定裝置[J];上海實驗動物科學(xué);1992年04期

3 肖柳英,林培英,馮昭明,張丹;不同周齡的SD大鼠生理、生化及體重的正常值測定[J];中藥新藥與臨床藥理;1996年03期

4 李淑云;簡易大鼠灌胃器的制作[J];錦州醫(yī)學(xué)院學(xué)報;2001年04期

5 楊明智,陳積圣;一種大鼠抓取與固定的新工具介紹[J];上海實驗動物科學(xué);2001年03期

6 戴英,陸群;復(fù)方H_(505)對Wistar大鼠外周血的血液流變學(xué)指標(biāo)的影響[J];中國血液流變學(xué)雜志;2001年01期

7 韋應(yīng)波,孫喜慶,曹新生,姚永杰,馮岱雅,楊長斌;+Gz暴露時間對大鼠記憶功能和行為的影響[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;2003年01期

8 呂學(xué)軍,郭俊生,李敏,周利梅,張永娟;暈船大鼠體內(nèi)鐵含量的變化[J];中國職業(yè)醫(yī)學(xué);2003年04期

9 湯仁仙,王迎偉,王慧,周峰;201A中藥合劑對大鼠抗腎小球基底膜腎炎病變的影響[J];徐州醫(yī)學(xué)院學(xué)報;2003年06期

10 孫同柱,付小兵,翁立新,梁雪梅,陳偉;介紹一種簡易的大鼠保定方法[J];上海實驗動物科學(xué);2004年01期

相關(guān)會議論文 前10條

1 尹音;孫振宇;胡敏;李冬霞;;持續(xù)性高正加速度對大鼠顳頜關(guān)節(jié)損傷的作用[A];第八屆全國顳下頜關(guān)節(jié)病學(xué)及（牙合）學(xué)大會論文匯編[C];2011年

2 祝~=驤;iJ梊霞;洃克琴;崔素英;文允摪;，

本文編號：1366437