豐富環(huán)境和游泳運動對自閉癥大鼠CX43、GSK3β、Bcl-2蛋白表達的影響
發(fā)布時間:2018-02-26 23:36
本文關鍵詞: 自閉癥 游泳運動 豐富環(huán)境 CX43 GSK3β Bcl-2 出處:《吉林大學》2017年碩士論文 論文類型:學位論文
【摘要】:目的:觀察豐富環(huán)境和游泳干預對自閉癥模型大鼠腦皮CX43、GSK3β、Bcl-2蛋白表達的調控。實驗方法:將雌、雄Wistar大鼠各1只置于同一籠內并過夜,當檢測到雌鼠排出在便盤或者一直留在陰道內的淡黃色或白色的半透明的陰栓記胚胎為第0.5天。將受孕成功的母鼠挑出單獨喂養(yǎng),并隨機將受孕雌鼠分成2組,一組使用0.85%的生理鹽水配VPA粉沫并制作成250mg/ml溶液,在胚胎12.5天的時候將該溶液注入受孕母鼠體內(600mg/kg),該組受孕母鼠生產下的仔鼠記錄為模型組,對模型組的仔鼠進行篩選,得到自閉癥模型大鼠;對照組是未進行任何干預的正常母鼠產下的正常仔鼠。本論文實驗分為五組:游泳+豐富環(huán)境組(SE+EE)、豐富環(huán)境組(EE)、自閉癥組(ASD)、游泳運動組(SE)、空白對照組(C)。游泳運動組于每天下午3:00——4:30在30±1℃的水中進行持續(xù)90分鐘的中等負荷運動訓練,并持續(xù)訓練8周。豐富環(huán)境組生活在長、寬、高為1m×0.6m×0.4m的透明飼養(yǎng)籠中,籠內有多種玩具,讓自閉癥大鼠在其中訓練。干預結束后,斷頭取腦,用免疫組化方法檢測大鼠腦皮層CX43、GSK3β、Bcl-2蛋白的表達程度,應用Image-Pro Plus多功能真彩色細胞圖像分析管理系統(tǒng)進行圖像采集分析,同時計算出CX43、GSK3β、Bcl-2蛋白的平均熒光強度(熒光強度/總面積),并做出對比分析。實驗所得數據用SPSS13.0統(tǒng)計軟件進行分析。實驗結果:1.體重對比:與正常組大鼠體重相比,VPA自閉癥模型組仔鼠在出生后的第7天、第14天、第21天,PN70.05,有明顯的不同;PN14和PN21均0.05,沒有明顯的不同。2.睜眼時間:與正常組仔鼠相比,VPA模型組仔鼠的睜眼時間要稍微晚一些,PN13、PN14和PN15天時,在統(tǒng)計學上都具有差異性(P0.05)。3.方向趨向性:自閉癥幼鼠較正常組幼鼠相比,其轉身180o所用的時間要長一些,在PN7、PN8和PN9天,P0.05,在統(tǒng)計學上都有差異性。4.強迫游泳:與正常大鼠相比較,在PN8、PN10天,自閉癥模型組大鼠的游泳能力明顯要弱,P≤0.05,具有統(tǒng)計學差異。5.CX43蛋白表達:與模型組比較,游泳+豐富環(huán)境組腦皮質CX43蛋白表達較多,P0.05,具有顯著差異;與游泳運動組比較,豐富環(huán)境組CX43蛋白表達多于游泳組,P0.05,具有顯著差異。6.GSK3β蛋白表達:與模型組比較,游泳+豐富環(huán)境組GSK3β蛋白表達多于模型組,P0.05,具有顯著差異;與豐富環(huán)境組比較,游泳組腦皮質GSK3β蛋白表達較多,P0.05,有統(tǒng)計學差異。7.Bcl-2蛋白表達:與模型組比較,游泳+豐富環(huán)境組腦皮質Bcl-2蛋白表達較少,P0.05,具有顯著差異;與游泳運動組比較,豐富環(huán)境組Bcl-2蛋白表達P0.05,沒有統(tǒng)計學差異。結論:豐富環(huán)境和游泳運動可以增加CX43、GSK3β蛋白在自閉癥模型大鼠腦皮層的表達,降低Bcl-2蛋白的表達。
[Abstract]:Objective: to observe the regulation of rich environment and swimming intervention on the expression of Bcl 2 protein of CX43 (GSK3 尾) Bcl 2 in rat cortex. Methods: one male and one male Wistar rats were placed in the same cage and overnight. When a light yellow or white translucent vaginal suppository was detected in the feces or in the vagina, the embryo was marked at day 0.5. The successful pregnant rats were singled out for feeding alone and were randomly divided into two groups. One group used 0.85% normal saline and VPA powder foam to make 250mg / ml solution, and injected the solution into the pregnant mice at 12.5 days after embryo. The offspring produced by the group were recorded as model group, and the offspring of the model group were screened. The rat model of autism was obtained. The control group was a normal newborn rat without any intervention. This study was divided into five groups: swimming rich environment group, rich environment group, autistic group, swimming exercise group, and blank control group. Swimming was carried out in five groups: swimming rich environment group, rich environment group, autistic group, swimming exercise group and blank control group. The exercise group received 90 minutes of moderate exercise training in 30 鹵1 鈩,
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