低硒對(duì)發(fā)育期大鼠腦中creb及核基質(zhì)蛋白影響的研究

1、低硒對(duì)發(fā)育期大鼠腦中CREB及核基質(zhì)蛋白影響的研究2001級(jí)碩士研究生耿義群導(dǎo)師徐小虎教授蘇敏教授摘要目的觀察低硒對(duì)發(fā)育期大鼠大腦皮層和海馬的脂質(zhì)過(guò)氧化物代謝的影響，通過(guò)對(duì)轉(zhuǎn)錄因子cAMP反應(yīng)元件結(jié)合蛋白（CREB）含量和活性的測(cè)定，檢測(cè)大腦皮層和海馬神經(jīng)元的功能，初步探討低硒對(duì)NMPs的影響。方法利用本實(shí)驗(yàn)小組已建立的人工飼料喂養(yǎng)的三代低硒SD大鼠模型，分為對(duì)照組和低硒組。取第三代出生后21天的大鼠，雌雄不拘，經(jīng)麻醉斷頭處死后迅速取腦

2、，在冰上分離大腦皮層和海馬組織。用硫代巴比妥酸化學(xué)法（TBARS）測(cè)定大腦皮層和海馬的脂質(zhì)代謝產(chǎn)物MDA的含量（n=20）。分別提取大腦皮層和海馬的總蛋白，Westernblot免疫印跡觀察CREB和Ser133磷酸化CREB（pCREB）的表達(dá)情況（n=16），進(jìn)一步用免疫組化的方法在組織石蠟切片觀察大腦皮層的各種類(lèi)型神經(jīng)元及在海馬各區(qū)錐體細(xì)胞的表達(dá)(n=6)。提取大腦NMPs，經(jīng)初步鑒定后，進(jìn)行Westernblot免疫印跡觀察pC

3、REB的表達(dá)(n=6)。用高效毛細(xì)管電泳（HPCE）的毛細(xì)管區(qū)帶電泳（CZE）模式進(jìn)行NMPs的分離，觀察其組成成分的改變。結(jié)果測(cè)定低硒組大腦皮層MDA含量明顯高于對(duì)照組（對(duì)照組＝0.310.13，低硒組的＝0.660.10，P0.05）；海馬的MDA含量低硒組明顯高于對(duì)照組（對(duì)照組＝0.420.11，低硒組＝0.890.066，P0.05）。提取大腦皮層和海馬的總蛋白經(jīng)Westernblot免疫印跡檢測(cè)，顯示低硒組CREB與pCREB

4、明顯低于對(duì)照組。免疫組化結(jié)果用平均灰度值表示CREB在大腦皮層（對(duì)照組＝119.628.35，低硒組＝144.6716.67，P0.05）和海馬CA1區(qū)（對(duì)照組＝123.136.74，低硒組＝150.1623.80，P0.05）的表達(dá)，發(fā)現(xiàn)低硒組和對(duì)照組的表達(dá)均有降低。用陽(yáng)萬(wàn)方數(shù)據(jù)TheEffectsofSeleniumDeficiencyDietonCREBNuclearMatrixProteinsinBrainofDevelopin

5、gRatPostgraduate:GengYiqunSupervis:Prof.XuXiaohuProf.SuMinAbstractObjectToobservethechangesoflipidperoxideinthectexhippocampusofdevelopingratinducedbyseleniumdeficiencydietstudytheneuronfunctionbythecontentactivitydetect

6、ionofcAMPResponseElementBindingProtein(CREB)inctexhippocampus.TostudythealterationofNMPscomponents.MaterialmethodGroupsofhealthyadultSDratswithequalmalefemaleatromwerefedwithseleniumdeficiencyseleniumadequatefood.Thethir

7、dgenerationat21daysofagewerekilledthectexhippocampusweredissectedonicerapidly.ThecontentsofMDAwereassayedby2thiobarbituricacid(TBA)test.TheexpressionofCREBpCREBwereexaminedbywesternblotintheextractedtotalproteinfromctexh

8、ippocampus.TheexpressionthelocationofCREBpCREBwerefurtherexpledindifferentkindofneuroninthesameregionofbrainbyparaffinbrainsectionimmunohistochemistry.TheexpressionofpCREBwasalsodetectedfromtheextractedNMPsofratbrainbywe

9、sternblot.ThecomponentoftheNMPsbetweentwogroupswereanalysedcomparedbyHighPerfmanceCapillaryElectrophisis(HPCE).ResultThecontentofMDAinthectexhippocampusofseleniumdeficiencygroupwasmuchhigherthanthatofthecontrolgroup(cont