CAJ | 학술논문

目的通过研究Bcl-2(B-cell lymphoma/Leukemia-2)高表达转基因小鼠的脊髓损伤模型,探寻Bcl-2抗神经元凋亡,减轻脊髓继发性损害的作用.方法将Bcl-2质粒转入小鼠的受精卵,培育Bcl-2转基因小鼠.转基因(A组)及同窝非转基因小鼠(B组)各9只随机(随机数字法)分为三组(1d,7 d,14 d),采用垂直打击脊髓(weight dropping,WD)方法,以2.5×3.0 g·cm致伤力致伤小鼠脊髓.另取3只非转基因小鼠设为假手术组(C组),仅打开椎板,暴露脊髓,不做打击.于术后1 d,7 d,14 d分别行:①小鼠后肢运动功能(BBB)评分;②HE染色观察;③原位末端标记法(TUNEL)染色.结果打击后所有小鼠均出现双侧的后肢瘫痪,两周内BBB评分,A组(5.45±0.15)和B组(5.05±0.35),差异无统计学意义,P=0.67;HE染色提示A组小鼠的脊髓损伤程度较B组鼠要轻;原位末端标记法(TUNEL)染色表明A组小鼠TUNEL阳性的神经元(3.67±2.08/HP)明显小于B组小鼠(12±4/HP),P=0.033;积分光密度(Integration Optic Density,IOD)值(458.33±112.7)A组也明显小于B组小鼠(2436.33±228.01),P=0.000,其差异均具有统计学意义;C组未见明显的TUNEL阳性神经元.结论 Bcl-2蛋白是中枢神经系统内在的,重要的抗损伤因子,过表达Bcl-2能够抑制损伤后神经元的凋亡,从而减轻脊髓的损伤.
목적 통과연구Bcl-2(B-cell lymphoma/Leukemia-2)고표체전기인소서적척수손상모형,탐심Bcl-2항신경원조망,감경척수계발성손해적작용.방법 장Bcl-2질립전입소서적수정란,배육Bcl-2전기인소서.전기인(A조)급동와비전기인소서(B조)각9지수궤(수궤수자법)분위삼조(1d,7 d,14 d),채용수직타격척수(weight dropping,WD)방법,이2.5×3.0 g·cm치상력치상소서척수.령취3지비전기인소서설위가수술조(C조),부타개추판,폭로척수,불주타격.우술후1 d,7 d,14 d분별행:①소서후지운동공능(BBB)평분;②HE염색관찰;③원위말단표기법(TUNEL)염색.결과 타격후소유소서균출현쌍측적후지탄탄,량주내BBB평분,A조(5.45±0.15)화B조(5.05±0.35),차이무통계학의의,P=0.67;HE염색제시A조소서적척수손상정도교B조서요경;원위말단표기법(TUNEL)염색표명A조소서TUNEL양성적신경원(3.67±2.08/HP)명현소우B조소서(12±4/HP),P=0.033;적분광밀도(Integration Optic Density,IOD)치(458.33±112.7)A조야명현소우B조소서(2436.33±228.01),P=0.000,기차이균구유통계학의의;C조미견명현적TUNEL양성신경원.결론 Bcl-2단백시중추신경계통내재적,중요적항손상인자,과표체Bcl-2능구억제손상후신경원적조망,종이감경척수적손상.
Objective To study the variables of behavioral function and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) figure in Bcl-2 transgenic (TG) mice and control mice after spinal cord injury (SCI), thus to find new ideas and ways for diagnosing and treating SCI. Method The genesis of Bcl-2 overexpression transgenic (TG) mice were produced by injection of Bcl-2 plasmid into the fertilized ova of mice.Nine Bcl-2 TG mice and nine control mice were subjected to SCI of moderate severity at T10, with the use of weight dropping (WD) method (impact force 2.5～3.0 g·cm). Up to 1 day , 7 days, and 14 days after SCI,functional deficits were evaluated with BBB scales, and the apoptosis of neurons was investigated by using TUNEL method. Another three mice of control group were only treated with laminectomy without SCI for comparison. Results The mean functional scores in the control mice were lower than those in the Bcl-2 TG mice, although the unpaired T -test revealed no significant differences. On the other hand, the number of TUNEL positive neurons and IOD(Integrated Optical Density)score in the Bcl-2 TG mice were both significantly lower than those in the control mice. Conclusions This experiment suggests that overexpression of Bcl-2 may suppress neuronal apoptosis after SCI. The Bcl-2 may be an important factor in relieving the damage within CNS after trauma.