摘要
目的 :研究 β 淀粉样蛋白 (Betaamyloidpritein ,Aβ)与载脂蛋白E4(ApolipoproteinE ,ApoE4)对神经元的共同作用 ,探讨老年性痴呆发病的细胞分子机制。材料与方法 :体外培养神经细胞 ,采用MTT比色法和免疫组化标记 ,结合图像分析技术 ,研究Aβ3 1 3 5和ApoE4对海马神经元存活和生长的作用。结果 :(1 )Aβ3 1 3 5(1 0 0 μmol/L) +ApoE4(1 0 μg/ml)和Aβ3 1 3 5(2 0 μmol/L) +ApoE4(1 0 μg/ml)的OD值 ,分别为 0 .1 97± 0 .0 2 1和 0 .1 91± 0 .0 2 4,明显低于对照组 0 .2 2 9± 0 .0 0 3 μm(P <0 .0 5 )。 (2 )这两组神经元胞体的最长径分别为 1 0 .0 7± 1 .98μm和 1 0 .0 1± 1 .68μm ;最短径分别为 6.40± 0 .77μm ,6.2 8± 0 .89μm ,明显低于对照组 1 2 .73± 3 .0 0 μm、7.0 5± 1 .0 4μm ,Aβ3 1 3 5组 1 2 .0 9± 2 .45 μm、7.0 1± 1 .0 2 μm ,最长径和最短径 (P <0 .0 5 ) ;(3 )两组神经元突起平均长度分别为 2 6.3 6± 7.73 μm和2 3 .86± 7.2 9μm ,明显低于对照组 3 0 .88± 9.79μm、2 8.3 4± 4.40 μm ,P <0 .0 5。 (4 )Aβ3 1 3 5(2 0 μmol/L)组的平均突起长度 (2 6.81± 5 .42 μm) ,明显低于对照组和ApoE4组 3 0 .60± 7.3 0 μm(P <0 .0 5 )。
Objective:To study the neurotoxic effect of Beta amyloid protein(Aβ) and E4 allele of apolipoprotein E(ApoE4) on the hippocampal neurons, providing pathologic mechanism for Alzheimer's disease. Methods: MTT assay, immunocytochemistry and morphometry were used to observe the survival and growth of hippocampal neurons in vitro. Results:(1) Forty eight hours after treatment, the average OD values of Aβ 31 35 (10 μmol/L)+ApoE4(10 μg/ml) and Aβ 31 35 (20 μmol/L)+ApoE4(10 μg/ml) were 0.197±0.021 μm, 0.191±0.024 μm, lower than that of the control group 0.229±0.003 μm, P<0.05,(2) Four days after treatment, the maximum and minimum diameters of neuronal soma in both Aβ 31 35 (10 μmol/L)+ApoE4(10 μg/ml) 10.07±1.98 μm, 6.40±0.77 μm, respectively and Aβ 31 35 20 μmol/L+ApoE4 (10 μg/ml) 10.01±1.68 μm, 6.28±0.89 μm, respectively were decreased significantly compared with those of control 12.73±3.00 μm, 7.05±1.04 μm, respectively, ApoE4 (12.60±1.87) μm, 7.05±0.99 μm, respectively], Aβ 31 35 (10 μmol/L) 12.09±2.45 μm, 7.01±1.02 μm, respectively and Aβ 31 35 (20 μmol/L) 12.01±2.48 μm, 7.04±0.77 μm, respectively groups(P<0.05). (3)The average neurite length of both Aβ 31 35 (10 μmol/L)+ApoE4(10 μg/ml) and Aβ 31 35 (20 μmol/L)+ApoE4(10 μg/ml) group , Aβ 31 35 (10 μmol/L) 12.09±2.45 μm, 7.01±1.02 μm, respectively and Aβ 31 35 (20 μmol/L) 12.01±2.48 μm, 7.04±0.77 μm, respectively groups(P<0.05). (3)The average neurite length of both Aβ 31 35 (10 μmol/L)+ApoE4(10 μg/ml) and Aβ 31 35 (20 μmol/L)+ApoE4(10 μg/ml) group [(26.36±7.73)μm, (23.86±7.29)μm, respectively] decreased significantly compared with those of control(30.88±9.79) μm, ApoE4(30.60±7.30) μm, Aβ 31 35 (10 μmol/L) (28.34±4.40) μm group(P<0.05). (4) The average neurite length of group Aβ 31 35 (20 μmol/L)(26.81±5.42) μm was decreased significantly compared with those of control and ApoE4 group(P<0.05). Conclusion: These results suggest that the neurotoxic effect of Aβ 31 35 +ApoE4 is higher than that of Aβ 31 35 alone. ApoE4 has no significant effect on survival and growth of the cultured hippocampal neurons, however, it may play a role with Aβ 31 35 in the aspect of neurotoxicity.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2001年第3期197-201,共5页
Chinese Journal of Anatomy
基金
国家自然科学基金 (No .3 9670 2 4 4 )
广东省自然科学资助课题
