不同時(shí)期有氧運(yùn)動(dòng)對(duì)大鼠腦衰老和額葉NCAM表達(dá)的影響

林雪靈,李 雪,鄧文騫,王璐,楊澎湃,張業(yè)廷,袁瓊嘉

(成都體育學(xué)院運(yùn)動(dòng)醫(yī)學(xué)系,四川 成都 610041)

不同時(shí)期有氧運(yùn)動(dòng)對(duì)大鼠腦衰老和額葉NCAM表達(dá)的影響

林雪靈,李 雪,鄧文騫,王璐,楊澎湃,張業(yè)廷,袁瓊嘉

(成都體育學(xué)院運(yùn)動(dòng)醫(yī)學(xué)系,四川 成都 610041)

目的：探討D-半乳糖致衰老的SD大鼠在衰老前和衰老過(guò)程中進(jìn)行有氧運(yùn)動(dòng)對(duì)其腦衰老和額葉NCAM表達(dá)的影響。方法：3月齡雄性SD大鼠60只隨機(jī)分成運(yùn)動(dòng)預(yù)處理組(SD組)、衰老組(ND組)、生理鹽水對(duì)照組(NNa組)、衰老過(guò)程中有氧運(yùn)動(dòng)干預(yù)組(NDs組),n=15。取材后,測(cè)定大腦皮質(zhì)的SOD、GSH-Px活性和MDA含量,Real-time PCR、Western blotting檢測(cè)額葉NCAM基因的表達(dá),免疫組織化學(xué)檢測(cè)其表達(dá)部位。結(jié)果：ND組大鼠較NNa組出現(xiàn)了嗜睡、食欲不振、行動(dòng)遲緩、毛色枯黃卷曲并大量脫落等衰老體征;ND組SOD、GSH-Px活性非常顯著性低于NNa組(P＜0.01),MDA含量非常顯著性多于NNa組(P＜0.01),SD組SOD、GSH-Px活性和MDA含量較ND組均無(wú)顯著性差異(P＞0.05),NDs組SOD、GSH-Px活性非常顯著性高于ND組(P＜0.01), MDA含量非常顯著性少于ND組(P＜0.01);ND組額葉NCAM基因的mRNA和蛋白質(zhì)表達(dá)水平非常顯著性低于NNa組(P＜0.01), SD組額葉NCAM基因的mRNA和蛋白質(zhì)表達(dá)水平較ND組無(wú)顯著性差異(P＞0.05),NDs組額葉NCAM基因的mRNA和蛋白質(zhì)表達(dá)水平非常顯著性高于ND組(P＜0.01),額葉NCAM的免疫產(chǎn)物呈棕褐色,陽(yáng)性表達(dá)主要位于額葉顆粒層細(xì)胞周緣的神經(jīng)元胞漿中。結(jié)論：D-半乳糖致大鼠腦衰老造模成功;運(yùn)動(dòng)預(yù)處理對(duì)大鼠腦衰老和額葉NCAM的表達(dá)均未產(chǎn)生積極作用;衰老過(guò)程中進(jìn)行有氧運(yùn)動(dòng)有效地延緩腦衰老,顯著上調(diào)額葉NCAM的表達(dá)。

有氧運(yùn)動(dòng);衰老;神經(jīng)細(xì)胞粘附分子;額葉;反轉(zhuǎn)錄-聚合酶鏈反應(yīng);免疫印跡法;免疫組織化學(xué);大鼠

衰老往往伴隨著認(rèn)知能力下降,記憶障礙和神經(jīng)退行性疾病的易感性增加[1]。衰老機(jī)制有DNA損傷學(xué)說(shuō)[2]、自由基學(xué)說(shuō)[3]、免疫學(xué)說(shuō)[4]、端粒學(xué)說(shuō)[5]、線粒體學(xué)說(shuō)[6]等,其中自由基學(xué)說(shuō)是目前較公認(rèn)的學(xué)說(shuō)[7]。額葉是空間辨別性學(xué)習(xí)和記憶的中樞之一,與學(xué)習(xí)和記憶密切相關(guān)[8],神經(jīng)細(xì)胞粘附分子(neural cell adhesion molecule,NCAM)參與了海馬、額葉等腦區(qū)的軸突生長(zhǎng)、神經(jīng)細(xì)胞遷移和分化、神經(jīng)元的損傷修復(fù)、突觸可塑性和信號(hào)傳導(dǎo)等神經(jīng)活動(dòng)[9,10]。國(guó)內(nèi)外大量實(shí)驗(yàn)表明適宜的有氧運(yùn)動(dòng)增強(qiáng)機(jī)體的抗氧化能力,延緩腦衰老[11]。本實(shí)驗(yàn)通過(guò)對(duì)D-半乳糖致衰老的SD大鼠在衰老前和衰老過(guò)程中分別進(jìn)行中等負(fù)荷的游泳運(yùn)動(dòng)干預(yù),觀察不同時(shí)期有氧運(yùn)動(dòng)對(duì)大鼠腦衰老和額葉NCAM表達(dá)的影響,從分子水平上探討有氧運(yùn)動(dòng)對(duì)腦衰老的影響機(jī)制,為研究運(yùn)動(dòng)與腦健康提供神經(jīng)分子生物學(xué)依據(jù)。

1 材料和方法

1.1 動(dòng)物模型和取材

SPF級(jí)3月齡雄性SD大鼠60只,體重400±20g,購(gòu)自成都達(dá)碩生物科技有限公司,生產(chǎn)許可：CSXK (川)2008-24。分籠飼養(yǎng),自由飲水進(jìn)食。適應(yīng)性喂養(yǎng)1周后隨機(jī)分成4組(見(jiàn)表1)：運(yùn)動(dòng)預(yù)處理組(SD組)、衰老組(ND組)、生理鹽水對(duì)照組(NNa組)、衰老過(guò)程中有氧運(yùn)動(dòng)干預(yù)組(NDs組)。

表1 大鼠分組和造模方案

大鼠在透明玻璃缸(160cm×60cm×110cm)內(nèi)游泳,水深80cm,水溫34±2℃。適應(yīng)性游泳3d(30 min/d)后正式運(yùn)動(dòng),SD組每日19：00進(jìn)行60min/d的游泳運(yùn)動(dòng),每周訓(xùn)練6d,共4周,之后進(jìn)行D-半乳糖注射;NDs組每日9：00注射D-半乳糖,19：00進(jìn)行60min/d的游泳運(yùn)動(dòng),每周訓(xùn)練6d,共6周。采用腹腔注射D-半乳糖6周,100mg/kg/d的注射劑量。根據(jù)大鼠的體重,D-半乳糖用生理鹽水稀釋成5%的濃度(2ml/kg/d)。同時(shí)NNa組采用相同的劑量和方法注射生理鹽水。按照造模設(shè)計(jì)時(shí)間取材,步驟如下：快速斷頭取腦,冰生理鹽水漂洗,濾紙拭干,置于冰上迅速切離額葉和其余大腦,分裝入凍存管后放入液氮罐內(nèi)保存。

1.2 自由基指標(biāo)檢測(cè)

測(cè)定大腦皮質(zhì)的蛋白總量、超氧化物歧化酶(superoxide dismutase,SOD)活性、谷胱甘肽過(guò)氧化物酶(glutathione peroxidase,GSH-Px)活性和丙二醛(malonaldehyde,MDA)含量。稱量大腦皮質(zhì),按W(g)組織塊重量/V(m l)生理鹽水為1/9加入冰生理鹽水,制備10%的組織勻漿。離心10-15min(4℃3,000rpm),用生理鹽水將上清稀釋成1%的組織勻漿,待測(cè)。主要試劑：考馬斯亮蘭蛋白測(cè)定試劑盒、SODWST-1法測(cè)定試劑盒、微量還原型谷胱甘肽測(cè)定試劑盒、丙二醛測(cè)定試劑盒。均嚴(yán)格按照南京建成生物工程研究所提供的試劑盒說(shuō)明書進(jìn)行操作。

1.3 額葉NCAM的real-time PCR檢測(cè)

利用Trizol法提取總RNA,檢測(cè)其在260nm和280nm的吸光度值,確定RNA的純度和濃度,進(jìn)行瓊脂糖凝膠電泳分析RNA的完整性。反轉(zhuǎn)錄反應(yīng)：采用iScriptTMcDNA Synthesis Kit(BIO-RAD)試劑盒,配置反轉(zhuǎn)錄反應(yīng)液：5×iScript reaction mix 4μl,iScript reverse transcriptase 1μl,Total RNA 1μl,Nuclease-free water補(bǔ)足20μl。反應(yīng)條件：25℃5min,42℃30min, 85℃5min,4℃保溫,1 cycle。PCR擴(kuò)增反應(yīng)：采用SsoAdvancedTMSYBR?Green Supermix(BIO-RAD)試劑盒,配制反應(yīng)液：1×SsoAdvancedTMSYBR?Green Supermix 5μl,上游引物(10uM)0.5μl,下游引物(10μM) 0.5μl,cDNA 100ng-100fg,RNase-free water補(bǔ)足10μl。反應(yīng)條件：95℃30sec 1 cycle,95℃5sec 41 cycle,56℃30sec 41 cycle,65℃-95℃5sec/step 1 cycle。PCR擴(kuò)增的寡核苷酸引物由TaKaRa寶生物工程有限公司設(shè)計(jì)合成(見(jiàn)表2)。實(shí)驗(yàn)數(shù)據(jù)采用2-△△ct法分析得出。

表2 引物序列及產(chǎn)物長(zhǎng)度

1.4 額葉NCAM的western blotting檢測(cè)

按1mg組織/20μl裂解液加入裂解液冰中勻漿,超聲破膜,冰中靜置30min,離心5min(4℃12, 000rpm),上清即為總蛋白樣品。在100℃水浴鍋進(jìn)行蛋白變性10min,冷卻后分裝,-20℃保存。采用碧云天BCA蛋白濃度測(cè)試試劑盒進(jìn)行蛋白濃度測(cè)定。取蛋白樣品進(jìn)行SDS-PAGE凝膠電泳分離,轉(zhuǎn)膜。將PVDF膜置于8%的脫脂奶粉中室溫封閉1h,孵育一抗(ab9018,用8%的脫脂奶粉稀釋,濃度1：1000)4℃過(guò)夜,37℃孵育二抗2h(山羊抗小鼠,lgG/辣根酶標(biāo)志,濃度1：2000),將顯影液用槍吹洗PVDF膜1min后用保鮮膜覆蓋,暗室內(nèi)壓片、顯影、定影。結(jié)果經(jīng)掃描后,采用Quantity One分析軟件,用條帶軌跡定量分析法對(duì)圖片上的條帶進(jìn)行蛋白表達(dá)的相對(duì)灰度值測(cè)量,以β-actin作內(nèi)參,分別計(jì)算各組額葉NCAM總蛋白的相對(duì)表達(dá)值。

1.5 額葉NCAM免疫組織化學(xué)SABC染色

常規(guī)石蠟包埋連續(xù)切片,厚4μm,60℃烤箱過(guò)夜。石蠟切片脫蠟至水,用3%H2O2室溫避光孵育15min,用高壓蒸汽鍋(129Kpa)進(jìn)行抗原修復(fù)3min,自然冷卻至室溫,滴加5%BSA置室溫20min,孵育一抗(兔IgG一抗/一抗稀釋液1：400)4℃過(guò)夜,孵育二抗(山羊抗兔IgG)置37℃烤箱20min,滴加SABC置37℃烤箱20min,DAB顯色,蘇木素復(fù)染15min,1%的鹽酸酒精分化25s,常規(guī)脫水、透明、封片。試劑均購(gòu)于武漢博士德生物工程有限公司。用Leica光學(xué)顯微鏡進(jìn)行圖像采集,400×光鏡下選取5個(gè)互不重疊同面積的視野。圖像用image pro-plus進(jìn)行處理,作背底校正后計(jì)積分光密度值。

1.6 統(tǒng)計(jì)學(xué)分析

用SPSS統(tǒng)計(jì)軟件包處理,計(jì)為平均值±標(biāo)準(zhǔn)差(mean±SD)。組間采用單因素方差分析(ANOVA),在P＜0.05和P＜0.01水平上分析顯著性差異。

2 結(jié)果

2.1 大鼠狀態(tài)觀察

NNa組大鼠精神狀態(tài)佳、飲食正常、行動(dòng)敏捷、毛色白有光澤;ND組大鼠出現(xiàn)了嗜睡、食欲不振、行動(dòng)遲緩、毛色枯黃卷曲并大量脫落等明顯的衰老體征。

2.2 自由基檢測(cè)結(jié)果

表3 各組大鼠大腦皮質(zhì)SOD、GSH-Px活性和MDA含量(M±SD,n=9)

2.3 額葉NCAM的Real-time PCR檢測(cè)結(jié)果

圖1 各組大鼠額葉NCAM基因的mRNA表達(dá)水平

2.4 額葉NCAM的Western blotting檢測(cè)結(jié)果

圖2 各組大鼠額葉NCAM基因的蛋白質(zhì)表達(dá)水平

圖3 各組大鼠額葉NCAM的免疫印跡條帶

2.5 額葉NCAM的免疫組織化學(xué)檢測(cè)結(jié)果

光鏡下(400倍)觀察切片可見(jiàn),NCAM的免疫產(chǎn)物呈棕褐色,陽(yáng)性表達(dá)主要存在于額葉顆粒層細(xì)胞周緣的神經(jīng)元胞漿中(見(jiàn)圖4)。

3 討論

3.1 大鼠腦衰老造模驗(yàn)證

采用公認(rèn)的D-半乳糖致衰老模型[12]。機(jī)體內(nèi)過(guò)量的D-半乳糖經(jīng)半乳糖氧化酶催化為醛糖和過(guò)氧化氫,產(chǎn)生超氧陰離子和氧衍生的自由基;或與蛋白質(zhì)或多肽中的游離胺反應(yīng),生成晚期糖基化終末產(chǎn)物(AGEs),致活性氧(ROS)積聚[13]。此外,過(guò)量的D-半乳糖可減少顆粒細(xì)胞層神經(jīng)元的遷移和新神經(jīng)元的生成[14],加速了膽堿能神經(jīng)元受損,免疫活動(dòng)減弱,基因表達(dá)異常,鈣穩(wěn)態(tài)失衡及線粒體功能障礙[15]等,產(chǎn)生與自然衰老相似體征。小鼠或大鼠可在頸背皮下、眼球后或腹腔注射D-半乳糖6-8周,劑量為50-500mg/Kg/d[16],建立衰老模型。Xing-Tai Li等(2010)對(duì)小鼠進(jìn)行頸背皮下注射D-半乳糖(100mg/ Kg/d)7周發(fā)現(xiàn),小鼠肝臟的SOD、GSH-Px活性顯著降低(P＜0.001)[17]。彭彬等(2011)報(bào)道了長(zhǎng)期低劑量注射D-半乳糖引起大鼠空間學(xué)習(xí)記憶能力下降,腦皮質(zhì)細(xì)胞的抗氧化能力降低等腦衰老表現(xiàn)[18]。本實(shí)驗(yàn)通過(guò)腹腔注射6周100mg/Kg/d的D-半乳糖建立大鼠衰老模型。大鼠狀態(tài)觀察發(fā)現(xiàn)：ND組大鼠較NNa組出現(xiàn)了嗜睡、食欲不振、行動(dòng)遲緩、毛色枯黃卷曲并大量脫落等明顯的自然衰老體征。自由基檢測(cè)結(jié)果顯示：ND組大鼠大腦皮質(zhì)的SOD、GSH-Px活性非常顯著性低于NNa組(P＜0.01),MDA含量非常顯著性多于NNa組(P＜0.01)。證明本實(shí)驗(yàn)D-半乳糖致大鼠腦衰老造模成功。

圖4 免疫組化顯示各組額葉NCAM表達(dá)和分布情況(×400)箭頭示陽(yáng)性細(xì)胞,SABC法免疫組化染色。標(biāo)尺示50μm

圖5 各組大鼠額葉NCAM免疫產(chǎn)物IOD值

3.2 不同時(shí)期有氧運(yùn)動(dòng)干預(yù)對(duì)大鼠腦衰老的影響

有氧運(yùn)動(dòng)并沒(méi)有抑制腦組織中自由基的產(chǎn)生[22],而主要是通過(guò)增強(qiáng)腦組織的抗氧化能力,加快對(duì)自由基的消除。而大強(qiáng)度無(wú)氧運(yùn)動(dòng)或力竭運(yùn)動(dòng)會(huì)減弱機(jī)體的抗氧化能力[11]。長(zhǎng)期適宜的有氧運(yùn)動(dòng)使機(jī)體的抗氧化防御體系對(duì)運(yùn)動(dòng)產(chǎn)生適應(yīng)性改變,增強(qiáng)免疫、神經(jīng)發(fā)生和突觸可塑性[19],減少星形膠質(zhì)細(xì)胞肥大和髓鞘失調(diào),調(diào)節(jié)腦血管內(nèi)皮生長(zhǎng)因子(VEGF)[20]、腦源性神經(jīng)營(yíng)養(yǎng)因子(BDNF)[23]和神經(jīng)細(xì)胞粘附分子(NCAM)[28]等表達(dá),增加抗炎細(xì)胞因子(IL10)水平[1]等,進(jìn)而延緩腦衰老。SOD是機(jī)體內(nèi)抗氧化系統(tǒng)的首要防御,將超氧陰離子轉(zhuǎn)成過(guò)氧化氫,GSH-Px清除脂類氫過(guò)氧化物,并在CAT含量很少或過(guò)氧化氫產(chǎn)量很低的組織中,可代替CAT清除過(guò)氧化氫[12]。MDA是脂質(zhì)過(guò)氧化的代謝產(chǎn)物,其含量反映機(jī)體脂質(zhì)過(guò)氧化水平和細(xì)胞受自由基攻擊的損傷程度[21]。Marosi K等(2012)通過(guò)對(duì)12月齡雌性Wistar大鼠進(jìn)行為期15周中等負(fù)荷的跑步運(yùn)動(dòng)發(fā)現(xiàn),運(yùn)動(dòng)組大鼠較對(duì)照組出現(xiàn)海馬ROS和蛋白羰基量減少,SOD、GSH-Px活性增強(qiáng),P-AMPK和PGC-1α上調(diào),表明長(zhǎng)期有氧運(yùn)動(dòng)可提高海馬的抗氧化能力,保護(hù)神經(jīng)元在衰老早期免受氧化應(yīng)激[24]。李垂坤(2011)研究發(fā)現(xiàn)60 min/d的游泳運(yùn)動(dòng)使小鼠大腦SOD活性升高,MDA含量減少,對(duì)延緩腦衰老起到有效的促進(jìn)作用[25]。本實(shí)驗(yàn)自由基檢測(cè)結(jié)果顯示：SD組大鼠大腦皮質(zhì)的SOD、GSH-Px活性和MDA含量較ND組無(wú)顯著性差異(P＞0.05); NDs組SOD、GSH-Px活性非常顯著性高于ND組(P＞0.01),MDA含量非常顯著性少于ND組(P＜0.01)。表明衰老過(guò)程中進(jìn)行有氧運(yùn)動(dòng)顯著增強(qiáng)了大鼠大腦皮質(zhì)的SOD、GSH-Px活性,減少了MDA含量,有效地延緩腦衰老,與大多文獻(xiàn)報(bào)道一致。但衰老前進(jìn)行有氧運(yùn)動(dòng)預(yù)處理對(duì)最終大鼠的腦衰老未產(chǎn)生積極作用,提示大鼠衰老前進(jìn)行有氧運(yùn)動(dòng)對(duì)機(jī)體所產(chǎn)生的積極作用可能會(huì)隨著運(yùn)動(dòng)的停止而逐漸消退。

3.3 不同時(shí)期有氧運(yùn)動(dòng)干預(yù)對(duì)大鼠額葉NCAM表達(dá)的影響

NCAM是一種單鏈膜結(jié)合糖蛋白,主要表達(dá)于中樞神經(jīng)系統(tǒng)細(xì)胞表面,尤其在海馬、額葉等腦區(qū)參與軸突生長(zhǎng)、信號(hào)傳導(dǎo)、神經(jīng)細(xì)胞遷移和分化、神經(jīng)元的損傷修復(fù)和突觸可塑性等神經(jīng)活動(dòng)[11,12]。Aonurm-Helm A(2008)研究發(fā)現(xiàn)缺失NCAM的腦區(qū)內(nèi)CREB介導(dǎo)的信號(hào)通路異常[26]。袁瓊嘉等(2012)報(bào)道了長(zhǎng)期中等負(fù)荷有氧運(yùn)動(dòng)提高了大鼠海馬NCAM的表達(dá)和空間學(xué)習(xí)記憶能力[27]。本研究結(jié)果顯示：額葉NCAM的免疫產(chǎn)物呈棕褐色,陽(yáng)性表達(dá)主要位于額葉顆粒細(xì)胞周緣的神經(jīng)元胞漿中。ND組額葉NCAM基因的mRNA和蛋白質(zhì)表達(dá)水平非常顯著性低于NNa組(P＜0.01),表明隨著大鼠腦衰老,額葉NCAM基因的mRNA和蛋白質(zhì)表達(dá)水平出現(xiàn)下降;SD組額葉NCAM基因的mRNA和蛋白質(zhì)表達(dá)水平較ND組無(wú)顯著性差異(P＞0.05),表明運(yùn)動(dòng)預(yù)處理未顯著上調(diào)額葉NCAM基因的mRNA和蛋白質(zhì)表達(dá)水平,提示大鼠衰老前進(jìn)行有氧運(yùn)動(dòng)對(duì)額葉NCAM基因表達(dá)的上調(diào)作用,可能會(huì)隨著運(yùn)動(dòng)的停止產(chǎn)生了NCAM的泛素化[28]等而逐漸減退,其機(jī)制有待于進(jìn)一步探討;NDs組額葉NCAM基因的mRNA和蛋白質(zhì)表達(dá)水平非常顯著性高于ND組(P＜0.01),與大多數(shù)文獻(xiàn)報(bào)道一致,表明衰老過(guò)程中有氧運(yùn)動(dòng)可顯著上調(diào)額葉NCAM基因的mRNA和蛋白質(zhì)表達(dá),促進(jìn)衰老大鼠神經(jīng)元的損傷修復(fù)和信號(hào)傳導(dǎo)等,維持其學(xué)習(xí)記憶能力。

4 結(jié)論

(1)D-半乳糖致大鼠腦衰老造模成功。

(2)運(yùn)動(dòng)預(yù)處理對(duì)大鼠腦衰老和額葉NCAM的表達(dá)均未產(chǎn)生積極作用。

(3)衰老過(guò)程中進(jìn)行有氧運(yùn)動(dòng)有效地延緩腦衰老,顯著上調(diào)額葉NCAM的表達(dá),促進(jìn)大鼠神經(jīng)元的損傷修復(fù)和信號(hào)傳導(dǎo)。

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Effect of aerobic exercise of different periods on rats brain aging and expression of NCAM gene in frontal lobe

LIN Xue-ling,et al
(Chengdu Sport University,Chengdu Sichuan 610041)

Objective：Aging SD rats induced by injection of D-Galactosewere intervened by aerobic exercise ahead of aging and during aging to explore the effectof aerobic exercise on brain aging and expression level of NCAM in frontal lobe.Methods：60 3-month-old male SD rats were randomly divided into groups SD(aerobic exercise ahead of aging group),ND(aging group),NNa(saline control group)and NDs(aerobic exercise during aging group),n=15.SOD activity,GSH-Px activity and MDA contents in cerebral cortex were determined.Expression level of NCAM in frontal lobewas determined by real-time PCR,western blotting and immunohistochemistry.Results：Group ND,compared with group NNa rats,revealed a lotof obvious aging signs,such as lethargy,loss of appetite,slowness,curly brown hair and shedding;SOD and GSH-Px activity of group ND ratswere significantly lower than group NNa(P＜0.01),MDA content was significantlymore than that of group NNa(P＜0.01),SOD and GSH-Px activity and MDA content of group ND rats had no significant difference from group ND(P＞0.05),SOD and GSH-Px activity of group NDs ratswere very significantly higher than those of group ND(P＜0.01),MDA contentwas significantly less than that of group ND(P＜0.01); mRNA and protein expression level of NCAM gene in frontal lobe of group ND ratswere significantly lower than those of group NNa(P＜0.01).Compared with group ND,mRNA and protein expression level of NCAM gene in frontal lobe of group SD rats had no significant difference(P＞0.05)whilemRNA and protein expression level of NCAM gene in frontal lobe of group NDs ratswere significantly higher than those of group ND(P＜0.01.Immunoreactivities of NCAM in frontallobe were brown,mainly located in neuronal cytoplasm beside the granular cells.Conclusion：Injection of D-galactose induced brain agingmodel rats successful;Aerobic exercise ahead of aging did not have a favorable impact on brain aging and aerobic exercise during aging showed an effective impact on delaying brain aging;Aerobic exercise ahead of aging did not have a favorable impact on expression level of NCAM in frontal lobe and aerobic exercise during aging could significantly increase expression level of NCAM in frontal lobe.

aerobic exercise;aging;neural cell adhesion molecule;frontal lobe;real-time PCR;Western blotting; immunohistochemistry;rat

G804.5

：A

：1001-9154(2014)06-0062-06

G804.5

：A

：1001-9154(2014)06-0062-06

國(guó)家自然科學(xué)基金(編號(hào)：31371202);四川省科技廳科技支撐計(jì)劃項(xiàng)目(編號(hào)：2012SZ0067);國(guó)家體育總局、四川省運(yùn)動(dòng)醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室;四川省高?？蒲袆?chuàng)新團(tuán)隊(duì)專項(xiàng)基金。

林雪靈(1989-),女,安徽宿州人,碩士研究生,主要研究方向?yàn)檫\(yùn)動(dòng)與健康促進(jìn)。

：袁瓊嘉。

2014-05-23