發(fā)布時間：2018-12-14 11:17

【摘要】：目的通過觀察戊四氮點燃大鼠模型海馬FAKpY397蛋白的動態(tài)表達(dá)變化,結(jié)合苔蘚纖維出芽的程度,探討其在苔蘚纖維出芽及癲癇致癇過程中的作用。 方法將135只健康的成年雄性Sprague-Dawley (SD)大鼠,隨機(jī)分為實驗組90只和對照組45只。實驗組每天腹腔注射亞驚厥劑量的PTZ(即戊四氮—35mg/kg,對照組每天腹腔注射同等量生理鹽水,成功點燃后停止注射(癇性發(fā)作等級評定標(biāo)準(zhǔn)參照Racine標(biāo)準(zhǔn))。根據(jù)首次腹腔注射后3天,1周,2周,4周和6周將實驗組及對照組分別隨機(jī)分為五個亞組。根據(jù)不同的實驗方法及用途再將每個不同的再亞組隨機(jī)細(xì)分為三個小組：一組用于免疫熒光及免疫組化染色,檢測各不同時間點實驗組與對照組FAKpY397蛋白在海馬的表達(dá)分布及表達(dá)量的變化；一組用于Western Blot,檢測不同時間點不同組別大鼠海馬區(qū)FAKpY397蛋白總表達(dá)量及其變化趨勢；另一組用于Timm染色,檢測海馬苔蘚纖維出芽情況并進(jìn)行評分。其中實驗組每小組各6只,對照組每小組各3只。 結(jié)果 1.動物行為學(xué) 隨著每天相同亞驚厥劑量PTZ經(jīng)腹腔注射給藥,實驗組大鼠所表現(xiàn)的癇性發(fā)作反應(yīng)逐漸加重,多數(shù)給藥后第21-28天(23.1±-2.3天)被成功點燃。PTZ致癇模型點燃率為93.75%,而死亡率為4.44%。被成功點燃的大鼠可見Racine I至RacineⅢ級的自發(fā)癇性發(fā)作。對照組大鼠未出現(xiàn)癇性發(fā)作,行為正常。 2.Western Blot結(jié)果顯示：在PTZ點燃致癇過程中,海馬FAKpY397蛋白總表達(dá)量于再PTZ腹腔注射后3d稍下降,1周時開始增加,4周達(dá)高峰,6周下降,仍高于對照組。除3天組外(0.05),實驗組各時間點與對照組間比較均有統(tǒng)計學(xué)差異(P0.05)。 3.免疫熒光及組化結(jié)果顯示：FAKpY397在海馬各區(qū)有表達(dá),主要分布于胞漿；免疫組化半定量分析示CA3區(qū)FAKpY397蛋白表達(dá)于PTZ腹腔注射后3d開始增加,4周達(dá)高峰,6周下降,仍高于對照組。實驗組中除1周組與2周組進(jìn)行比較無統(tǒng)計學(xué)差異(P0.05),其余各時間點兩兩比較及實驗組與對照組對應(yīng)時間點比較均有統(tǒng)計學(xué)差異(P0.05)。 4.苔蘚纖維出芽評分 對照組CA3區(qū)未見苔蘚纖維出芽或僅見輕微出芽,苔蘚纖維出芽評分在各時間點間兩兩比較均無統(tǒng)計學(xué)差異。實驗組CA3區(qū)苔蘚纖維出芽評分從PTZ腹腔注射后3天開始增加,4周時達(dá)高峰,6周下降。實驗組中除4周與6周間比較無統(tǒng)計學(xué)差異(0.05),其余各時間點兩兩比較及實驗組與對照組對應(yīng)時間點比較均有統(tǒng)計學(xué)差異(P0.05)。 結(jié)論 FAKpY397蛋白在海馬CA3區(qū)表達(dá)增多可能對于苔蘚纖維出芽具有促進(jìn)作用進(jìn)而誘發(fā)癲癇發(fā)作。
[Abstract]:Objective to investigate the role of pentylenetetrazol (PTZ) in the process of mossy fiber sprouting and epilepsy by observing the dynamic expression of FAKpY397 protein in hippocampus of rats with pentylenetetrazol (PTZ) kindling and combining with the degree of mossy fiber sprouting. Methods 135 healthy male Sprague-Dawley (SD) rats were randomly divided into experimental group (n = 90) and control group (n = 45). Subconvulsant dose of PTZ (pentylenetetrazo-35 mg / kg) was injected intraperitoneally every day in the experimental group, and the control group was injected with the same amount of normal saline daily. The injection was stopped after successful kindling (Racine criterion was used to evaluate the grade of epileptic seizures). According to 3 days, 1 week, 2 weeks, 4 weeks and 6 weeks after the first intraperitoneal injection, the experimental group and the control group were randomly divided into five subgroups. Each subgroup was subdivided randomly into three groups according to different experimental methods and uses: one group was used for immunofluorescence and immunohistochemical staining. The expression distribution and quantity of FAKpY397 protein in hippocampus of experimental group and control group were detected at different time points. One group was used to detect the total expression of FAKpY397 protein and its changing trend at different time points in different groups of rats, the other group was used for Timm staining to detect and score the budding of mossy fibers in hippocampus. There were 6 rats in each group in the experimental group and 3 rats in the control group. Result 1. With the same daily subconvulsive dose of PTZ administered intraperitoneally, the epileptic seizure response in the experimental group increased gradually. Most of the drugs were ignited 21-28 days (23.1 鹵-2.3 days) after administration. The kindling rate of epileptic model induced by PTZ was 93.755.The death rate was 4.44g. Spontaneous epileptic seizures of Racine I to Racine 鈪，

本文編號：2378524