本文選題：神經(jīng)元　切入點(diǎn)：星形膠質(zhì)細(xì)胞　出處：《重慶醫(yī)科大學(xué)》2012年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的無(wú)鎂誘導(dǎo)體外培養(yǎng)神經(jīng)元（neurons，N）放電，研究放電后N和星形膠質(zhì)細(xì)胞(astrocytes，AST)共培養(yǎng)體系中腦源性神經(jīng)營(yíng)養(yǎng)因子（brain-derived neurotrophic factor， BDNF）、神經(jīng)生長(zhǎng)因子（nerve growth factor，NGF）、谷氨酸（glutamate，GLU）、白介素-1beta（interleukin-1beta,IL-1β）動(dòng)態(tài)變化及乳酸脫氫酶（lacticdehydrogenase，LDH）含量。 方法以純化的胎鼠海馬N和新生鼠AST為研究對(duì)象，按照是否經(jīng)過(guò)短暫的無(wú)鎂標(biāo)準(zhǔn)細(xì)胞外液處理誘導(dǎo)反復(fù)的自發(fā)性驚厥樣放電，分為N對(duì)照組、N無(wú)鎂組、N+AST對(duì)照組和N+AST無(wú)鎂組。 ELISA方法測(cè)定驚厥后不同時(shí)間點(diǎn)細(xì)胞培養(yǎng)上清液中的BDNF、NGF、IL-1β含量；高效液相色譜法測(cè)定不同條件下各時(shí)間點(diǎn)細(xì)胞上清液中GLU含量；細(xì)胞培養(yǎng)上清液中LDH的變化采用全自動(dòng)生化分析儀測(cè)定。 結(jié)果（1）N經(jīng)無(wú)鎂細(xì)胞外液處理，恢復(fù)正常培養(yǎng)72h后，仍舊存在反復(fù)的自發(fā)性“驚厥樣放電”。（2）N+AST經(jīng)無(wú)鎂處理后48h，AST的體積增大，突起及細(xì)胞數(shù)目增多。（3）N+AST對(duì)照組在24h、48h細(xì)胞上清液BDNF較N對(duì)照組24h、48h細(xì)胞上清液BDNF升高（p㩳0.05）。N+AST經(jīng)無(wú)鎂處理后，在12h、24h、48h分泌的BDNF均較N+AST對(duì)照組相同時(shí)間點(diǎn)的BDNF升高，尤其12h、24h升高明顯（p㩳0.01）；而N經(jīng)同樣的無(wú)鎂細(xì)胞外液處理后，各個(gè)時(shí)間點(diǎn)BDNF含量也有升高，但與N對(duì)照組比較，差異無(wú)統(tǒng)計(jì)學(xué)意義。N+AST經(jīng)無(wú)鎂處理后在24h分泌的BDNF較N無(wú)鎂組24h分泌的BDNF升高（p㩳0.05）。（4）N+AST經(jīng)無(wú)鎂處理后，各個(gè)時(shí)間點(diǎn)NGF的分泌量與共培養(yǎng)對(duì)照組相同時(shí)間點(diǎn)相比均沒(méi)有發(fā)現(xiàn)明顯變化，而且各個(gè)時(shí)間點(diǎn)間NGF的含量也趨于平穩(wěn)。（5）N+AST經(jīng)無(wú)鎂標(biāo)準(zhǔn)細(xì)胞外液處理后，細(xì)胞上清液中GLU的含量從6h開(kāi)始就有增加，，在48h、72h含量均較對(duì)照組同時(shí)間點(diǎn)GLU含量顯著性增高，差異有統(tǒng)計(jì)學(xué)意義（p㩳0.05）。（6）N+AST經(jīng)過(guò)無(wú)鎂誘導(dǎo)反復(fù)自發(fā)的驚厥樣放電后，細(xì)胞上清液中IL-1β的含量在12h、24h、48h較對(duì)照組相同時(shí)間點(diǎn)IL-1β的含量顯著性增高，差異具有統(tǒng)計(jì)學(xué)意義（p㩳0.05，p㩳0.01）。（7）N+AST共培養(yǎng)體系經(jīng)無(wú)鎂標(biāo)準(zhǔn)細(xì)胞外液處理后，72h細(xì)胞上清液中乳酸脫氫酶顯著高于N+AST對(duì)照組（p㩳0.01）。 結(jié)論（1）N+AST在正常培養(yǎng)情況下分泌的BDNF由N和AST共同參與，N可能起主要分泌作用。短暫無(wú)鎂處理的N+AST，經(jīng)誘導(dǎo)放電后BDNF的分泌顯著增多，并且可能主要來(lái)自驚厥微環(huán)境活化的AST。（2）無(wú)鎂誘導(dǎo)N放電對(duì)共培養(yǎng)體系中NGF的分泌影響較小。（3）無(wú)鎂誘導(dǎo)N放電后與AST共培養(yǎng)微環(huán)境中GLU和IL-1β的分泌均顯著性增多，并且IL-1β的變化比GLU出現(xiàn)較早。（4）體外驚厥微環(huán)境對(duì)N具有損傷作用，導(dǎo)致LDH漏出增多。
[Abstract]:Objective to induce neuronal neuronsn (NN) discharges in cultured neurons without magnesium. To study the dynamic changes of brain-derived neurotrophic factor (BDNF), brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), nerve growth factor (NGF), glutamate glutamate (GLU), interleukin-1 beta (IL-1 尾) and the content of lactate dehydrogenase (LDH) in the co-cultured system of N and astrocytes. Methods the purified fetal rat hippocampal N and newborn rat AST were used to induce recurrent spontaneous convulsion-like discharges according to whether they were treated with transient magnesium free standard extracellular fluid. They were divided into N control group, N mg free group, N AST control group and N AST without magnesium group. ELISA method was used to determine the content of BDNF NGF FN IL-1 尾 in the supernatant of cell culture at different time points after convulsion, and high performance liquid chromatography (HPLC) was used to determine the content of GLU in cell supernatant at different time points after convulsion. The changes of LDH in supernatant of cell culture were measured by automatic biochemical analyzer. Results after treated with Mg-free extracellular solution for 72 h, there was still repeated spontaneous "convulsive discharges". The volume of AST was increased 48 h after treatment without magnesium. The cell supernatant BDNF in the control group was significantly higher than that in the control group at 24 h or 48 h. The BDNF in the supernatant of the cell supernatant was significantly higher than that in the control group at 24 h or 48 h. After mg free treatment, the BDNF secreted by 0.05U 路N AST at 12h ~ 24h was significantly higher than that of N AST control group at the same time, especially at 12h / 24h. After treated with the same magnesium free extracellular solution, the content of BDNF increased at different time points, but there was no significant difference compared with the control group. The BDNF secreted by N AST at 24 hours after magnesium free treatment was higher than that of BDNF secreted at 24 hours in N mg free group. There was no significant change in the secretion of NGF at each time point after mg free treatment of 0.05N AST, and the content of NGF at each time point tended to be stable after treated with magnesium free standard extracellular fluid. The content of GLU in the supernatant was increased from 6 h to 72 h, and the content of GLU in the supernatant was significantly higher than that in the control group at 48 h and 72 h, and the difference was statistically significant. After repeated spontaneous convulsion-like discharges induced by mg free, the content of IL-1 尾 in cell supernatant was significantly higher than that in control group at 12h ~ 24h ~ 48h, and the difference was statistically significant. 0.05,p? The lactate dehydrogenase (LDH) in the supernatant of 0. 01 AST / 0. 01% N AST was significantly higher than that in the N AST control group after being treated with magnesium free standard extracellular fluid for 72 h. 0.01g. Conclusion the BDNF secreted by N AST in normal culture may play a major role in the secretion of N by N and AST. The secretion of BDNF after induced discharge of N ASTs treated with transient mg free treatment is significantly increased. [WT5HZ] [WT5 "BZ] [WT5" BZ] [WT5 "BZ]. And it may be that the activation of AST.2from convulsion microenvironment has little effect on the secretion of NGF in co-culture system, and that the secretion of GLU and IL-1 尾 in microenvironment of co-culture with AST is significantly increased after Mg-free N discharges, and the effect of Mg-free N discharges on the secretion of GLU and IL-1 尾 in co-cultured microenvironment is significant. The change of IL-1 尾 was earlier than that of GLU.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 胡越;蔣莉;張曉萍;;持續(xù)驚厥后海馬神經(jīng)營(yíng)養(yǎng)因子表達(dá)及其影響因素[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2006年16期

2 李平,李正莉,朱長(zhǎng)庚,劉慶瑩,于珊,魏瑛;白細(xì)胞介素-1β對(duì)Fos蛋白表達(dá)的影響及其在癲癇發(fā)作中的作用[J];中國(guó)組織化學(xué)與細(xì)胞化學(xué)雜志;2003年03期

3 姚君茹,潘三強(qiáng),呂來(lái)清,高秀來(lái);慢性癲癇模型大鼠腦谷氨酸神經(jīng)元的變化[J];解剖科學(xué)進(jìn)展;2004年01期

4 王磊;;星形膠質(zhì)細(xì)胞在神經(jīng)系統(tǒng)疾病中的作用[J];生物技術(shù)通報(bào);2010年06期

5 蔣莉,蔡方成,李欣,張曉萍;驚厥持續(xù)狀態(tài)中大鼠腦內(nèi)神經(jīng)元死亡的年齡特征[J];中華兒科雜志;2002年09期

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