基底前腦巢蛋白陽(yáng)性及陰性膽堿能神經(jīng)元自然衰老及抗損傷的研究
發(fā)布時(shí)間：2018-07-28 17:57
【摘要】：研究背景基底前腦一般指位于大腦半球內(nèi)側(cè)面一些灰質(zhì)結(jié)構(gòu),與其他腦區(qū)無(wú)明顯界限。基底前腦是一個(gè)具有高度異質(zhì)性的區(qū)域,含有多種不同化學(xué)屬性的神經(jīng)元,主要包括膽堿能神經(jīng)元、谷氨酸能神經(jīng)元以及r-氨基丁酸神經(jīng)元等。最近研究發(fā)現(xiàn)成年大鼠和成人基底前腦存在著一群巢蛋白免疫陽(yáng)性(nestin-ir)神經(jīng)元,并證實(shí)這群神經(jīng)元屬于膽堿能神經(jīng)元。進(jìn)一步研究發(fā)現(xiàn)巢蛋白免疫陽(yáng)性(nestin-ir)膽堿能神經(jīng)元對(duì)學(xué)習(xí)記憶及對(duì)中樞神經(jīng)系統(tǒng)的損傷與修復(fù)起重要作用。國(guó)內(nèi)外初步研究提示相對(duì)于巢蛋白陰性膽堿能神經(jīng)元,基底前腦的巢蛋白免疫陽(yáng)性膽堿能神經(jīng)元可能具有更強(qiáng)的抗損傷能力,表現(xiàn)出更好的神經(jīng)可塑性。目的本課題采用基底前腦神經(jīng)元原代培養(yǎng)方法,通過(guò)細(xì)胞免疫熒光染色研究巢蛋白免疫陽(yáng)性及陰性膽堿能神經(jīng)元抗衰老及抗氧化損傷能力的不同,進(jìn)一步揭示兩種神經(jīng)元的神經(jīng)可塑性的差異,為相關(guān)的神經(jīng)系統(tǒng)疾病的治療來(lái)提供新基礎(chǔ)。方法實(shí)驗(yàn)采用出生24h以內(nèi)的新生SD大鼠,每次用量8只左右,取其基底前腦處的神經(jīng)元進(jìn)行體外培養(yǎng),培養(yǎng)周期11天,分別在培養(yǎng)的第1天,第3天,第5天,第7天,第9天和第11天,取培養(yǎng)的基底前腦神經(jīng)元細(xì)胞爬片進(jìn)行免疫熒光實(shí)驗(yàn),研究巢蛋白免疫陽(yáng)性及免疫陰性膽堿能神經(jīng)元的自然衰老過(guò)程�？箵p傷的研究采用出生24h以內(nèi)的新生SD大鼠,每次用量8只左右,取其基底前腦部位的神經(jīng)元進(jìn)行體外培養(yǎng),在培養(yǎng)的第3天時(shí),放入150μmol/L H2O2構(gòu)建細(xì)胞損傷模型,作用12h后,取出培養(yǎng)的基底前腦神經(jīng)細(xì)胞爬片進(jìn)行免疫熒光實(shí)驗(yàn),觀察兩種神經(jīng)元得抗損傷能力差異。結(jié)果體外培養(yǎng)第三天和第五天細(xì)胞生長(zhǎng)達(dá)到高峰,這時(shí)神經(jīng)元胞體較大,神經(jīng)元突起粗大,部分神經(jīng)元之間形成網(wǎng)狀的突觸連接。其中可見(jiàn)巢蛋白陽(yáng)性神經(jīng)元胞體較大,突起粗大且長(zhǎng);巢蛋白陰性神經(jīng)元胞體小,突起較少。體外培養(yǎng)五天之后的神經(jīng)元細(xì)胞數(shù)目逐漸減少,胞體飽和度降低。但巢蛋白陽(yáng)性神經(jīng)元數(shù)目下降速率緩慢,占細(xì)胞總數(shù)的比例逐漸增加,巢蛋白陰性神經(jīng)元數(shù)目下降速率較快,所占細(xì)胞總數(shù)的比例逐漸下降。正常衰老組從培養(yǎng)第7天開始巢蛋白陽(yáng)性及陰性膽堿能神經(jīng)元數(shù)目差異明顯,用T檢驗(yàn)方法,P0.05,有統(tǒng)計(jì)學(xué)意義。這表明在相同的培養(yǎng)條件下,巢蛋白免疫陽(yáng)性和陰性膽堿能神經(jīng)元自然衰老和死亡速度是不同,巢蛋白免疫陽(yáng)性膽堿能神經(jīng)元細(xì)胞數(shù)目下降的速率比巢蛋白陰性膽堿能神經(jīng)元慢。在神經(jīng)元抗氧化損傷實(shí)驗(yàn)中,用T檢驗(yàn)方法分析正常組和損傷組的巢蛋白陽(yáng)性及陰性膽堿能神經(jīng)元發(fā)現(xiàn),正常組中兩種神經(jīng)元數(shù)目無(wú)明顯差異,P0.05,無(wú)統(tǒng)計(jì)學(xué)意義。而損傷組中神經(jīng)元P0.05,有統(tǒng)計(jì)學(xué)意義。這充分說(shuō)明H2O2氧化損傷干預(yù)后巢蛋白陽(yáng)性膽堿能神經(jīng)元存活的數(shù)目較巢蛋白陰性膽堿能神經(jīng)元多,細(xì)胞形態(tài)更好,比巢蛋白陰性膽堿能神經(jīng)元有更強(qiáng)的抗損傷能力。結(jié)論體外培養(yǎng)的基底前腦神經(jīng)元自然衰老及氧化損傷研究結(jié)果表明,與巢蛋白陰性膽堿能神經(jīng)元相比,基底前腦的巢蛋白陽(yáng)性膽堿能神經(jīng)元抗衰老和抗損傷能力更強(qiáng),表現(xiàn)出較強(qiáng)的神經(jīng)可塑性。
[Abstract]:Background basal forebrain usually refers to some gray matter in the inner hemisphere of the cerebral hemisphere and has no distinct boundary with other brain regions. The basal forebrain is a highly heterogeneous region with a variety of neurons with different chemical properties, including cholinergic neurons, glutamic acid neurons and r- aminobutyric acid neurons. It was found that a group of nestin immunoreactive (Nestin-IR) neurons were found in the adult rat and adult basal forebrain, and confirmed that these neurons belong to cholinergic neurons. Further studies have found that nestin immunoreactive (Nestin-IR) cholinergic neurons play an important role in learning and memory and the damage and repair of the central nervous system. The preliminary study suggests that the nestin immunoreactive cholinergic neurons in the basal forebrain may have a stronger ability to resist damage and exhibit better neuroplasticity relative to nestin negative cholinergic neurons. The difference between the anti aging and antioxidant damage of the positive and negative cholinergic neurons further reveals the difference between the nerve plasticity of the two kinds of neurons and provides a new basis for the treatment of the related nervous system diseases. Methods the new SD rats under the birth 24h were used for each dose of about 8, and the nerve in the basal forebrain was taken. The cells were cultured in vitro and cultured for 11 days. The cultured basal forebrain neuron cells were taken for immunofluorescence test at first days, third days, fifth days, seventh days, Ninth days and eleventh days respectively, and the natural aging process of nestin immunoreactive and immune negative cholinergic neurons was studied. The anti injury study was born 24h. In the newborn SD rats, the neurons of the basal forebrain were cultured in vitro for about 8 rats each time. At the third day of the culture, 150 mu mol/L H2O2 was put into the cell damage model. After the action of 12h, the cultured basal forebrain nerve cell crawling slices were taken for immunofluorescence experiment, and the differences of the anti injury ability of the two kinds of neurons were observed. Results the cell growth reached the peak at third days and fifth days in vitro. At this time, the cell body was larger, the neurites were large, and the neurons formed a network of synapses. Among them, the nestin positive neurons were larger, the protuberances were large and long, and the nestin negative neurons were small, and the protuberances were less. In vitro culture, the cells were cultured for five days. The number of neuron cells decreased gradually and the cell saturation decreased, but the number of nestin positive neurons decreased slowly, the proportion of the total number of cells increased gradually, the number of nestin negative neurons decreased rapidly and the proportion of the total number of cells decreased gradually. The normal senescence group began to be nestin positive and negative from seventh days of culture. The difference of the number of sexual cholinergic neurons was obvious. The T test method, P0.05, had statistical significance. This showed that under the same culture conditions, the natural aging and death rate of nestin positive and negative cholinergic neurons were different, and the rate of nestin positive cholinergic neurons decreased more than the nestin negative cholinergic neurons. The T test was used to analyze the nestin positive and negative cholinergic neurons in the normal group and the injured group by T test. There was no significant difference in the number of neurons in the normal group, and there was no significant difference in the number of neurons in the normal group, P0.05, but the neuron P0.05 in the injured group was statistically significant. This fully explained the oxidative damage of H2O2. After intervention, the number of nestin positive cholinergic neurons survives more than nestin negative cholinergic neurons, and the cell morphology is better than nestin negative cholinergic neurons. Conclusion the results of natural senescence and oxidative damage of the basal forebrain neurons in vitro show that the nestin negative cholinergic neurons are the same as nestin negative cholinergic. In contrast, the nestin positive cholinergic neurons in basal forebrain were more resistant to aging and injury, showing stronger neuroplasticity.
【學(xué)位授予單位】：大理大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R741
本文編號(hào)：2151148
[Abstract]:Background basal forebrain usually refers to some gray matter in the inner hemisphere of the cerebral hemisphere and has no distinct boundary with other brain regions. The basal forebrain is a highly heterogeneous region with a variety of neurons with different chemical properties, including cholinergic neurons, glutamic acid neurons and r- aminobutyric acid neurons. It was found that a group of nestin immunoreactive (Nestin-IR) neurons were found in the adult rat and adult basal forebrain, and confirmed that these neurons belong to cholinergic neurons. Further studies have found that nestin immunoreactive (Nestin-IR) cholinergic neurons play an important role in learning and memory and the damage and repair of the central nervous system. The preliminary study suggests that the nestin immunoreactive cholinergic neurons in the basal forebrain may have a stronger ability to resist damage and exhibit better neuroplasticity relative to nestin negative cholinergic neurons. The difference between the anti aging and antioxidant damage of the positive and negative cholinergic neurons further reveals the difference between the nerve plasticity of the two kinds of neurons and provides a new basis for the treatment of the related nervous system diseases. Methods the new SD rats under the birth 24h were used for each dose of about 8, and the nerve in the basal forebrain was taken. The cells were cultured in vitro and cultured for 11 days. The cultured basal forebrain neuron cells were taken for immunofluorescence test at first days, third days, fifth days, seventh days, Ninth days and eleventh days respectively, and the natural aging process of nestin immunoreactive and immune negative cholinergic neurons was studied. The anti injury study was born 24h. In the newborn SD rats, the neurons of the basal forebrain were cultured in vitro for about 8 rats each time. At the third day of the culture, 150 mu mol/L H2O2 was put into the cell damage model. After the action of 12h, the cultured basal forebrain nerve cell crawling slices were taken for immunofluorescence experiment, and the differences of the anti injury ability of the two kinds of neurons were observed. Results the cell growth reached the peak at third days and fifth days in vitro. At this time, the cell body was larger, the neurites were large, and the neurons formed a network of synapses. Among them, the nestin positive neurons were larger, the protuberances were large and long, and the nestin negative neurons were small, and the protuberances were less. In vitro culture, the cells were cultured for five days. The number of neuron cells decreased gradually and the cell saturation decreased, but the number of nestin positive neurons decreased slowly, the proportion of the total number of cells increased gradually, the number of nestin negative neurons decreased rapidly and the proportion of the total number of cells decreased gradually. The normal senescence group began to be nestin positive and negative from seventh days of culture. The difference of the number of sexual cholinergic neurons was obvious. The T test method, P0.05, had statistical significance. This showed that under the same culture conditions, the natural aging and death rate of nestin positive and negative cholinergic neurons were different, and the rate of nestin positive cholinergic neurons decreased more than the nestin negative cholinergic neurons. The T test was used to analyze the nestin positive and negative cholinergic neurons in the normal group and the injured group by T test. There was no significant difference in the number of neurons in the normal group, and there was no significant difference in the number of neurons in the normal group, P0.05, but the neuron P0.05 in the injured group was statistically significant. This fully explained the oxidative damage of H2O2. After intervention, the number of nestin positive cholinergic neurons survives more than nestin negative cholinergic neurons, and the cell morphology is better than nestin negative cholinergic neurons. Conclusion the results of natural senescence and oxidative damage of the basal forebrain neurons in vitro show that the nestin negative cholinergic neurons are the same as nestin negative cholinergic. In contrast, the nestin positive cholinergic neurons in basal forebrain were more resistant to aging and injury, showing stronger neuroplasticity.
【學(xué)位授予單位】：大理大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R741
【參考文獻(xiàn)】
相關(guān)期刊論文 前5條
1 朱丹青;任新;王戚君;尹海燕;王欽;朱建華;;基底前腦巢蛋白免疫反應(yīng)陽(yáng)性神經(jīng)元的來(lái)源探索[J];解剖學(xué)研究;2014年01期
2 Yuhong Zhao;Kaihua Guo;Dongpei Li;Qunfang Yuan;Zhibin Yao;;Special function of nestin~+ neurons in the medial septum-diagonal band of Broca in adult rats[J];Neural Regeneration Research;2014年03期
3 李東培;汪華僑;徐杰;袁群芳;姚志彬;;大鼠基底前腦Nestin-IR神經(jīng)元的纖維投射[J];解剖科學(xué)進(jìn)展;2006年04期
4 周立兵,阮奕文,姚志彬;秋水仙素對(duì)大鼠基底前腦Nestin陽(yáng)性神經(jīng)元的影響[J];解剖科學(xué)進(jìn)展;2003年02期
5 周麗華,姚志彬,陳以慈;基底前腦解剖學(xué)及其臨床意義[J];神經(jīng)解剖學(xué)雜志;1998年04期
，本文編號(hào)：2151148
本文鏈接：http://sikaile.net/yixuelunwen/shenjingyixue/2151148.html
最近更新
教材專著
