摘要
目的:探讨重组人睫状神经营养因子(rhCNTF)对Aβ损伤的海马神经元的作用机制。方法:分离、纯化Wistar乳鼠海马组织,获得海马神经元并进行体外培养。采用神经元特异性烯醇化酶(NSE)免疫细胞化学进行鉴定。100 ng·mL-1的rhCNTF和10μmol·L-1的聚集态Aβ作用于海马神经元。CCK-8法检测rhCNTF对Aβ损伤的海马神经元活力影响。流式细胞术检测rhCNTF对Aβ致海马神经元早期凋亡的影响。结果 :免疫细胞化学法成功鉴定了海马神经元。海马神经元活动度随着rhCNTF浓度增加而增大且呈指数相关。镜下观察到rhCNTF+Aβ组细胞形态较Aβ组好;相对于Aβ组,rhCNTF+Aβ组和Aβ+rhCNTF组的海马神经元活动度显著增高(P<0.01)。rhCNTF可减少Aβ引起的细胞早期凋亡。结论 :rhCNTF对Aβ损伤的海马神经元有保护作用,其机制与促进神经元生长和减少细胞凋亡有关。
Objective： To investigate the mechanism of recombinant human ciliary neurotrophic factor （rhCNTF） onprimary cultured hippocampal neurons injured by β-amyloid fragment （Aβ）. Methods： Hippocampal neurons isolated andpurified from hippocampus in new-born Wistar rats were incubated in vitro, and identified by the neuron-specific enolase （NSE） ofimmunocytochemistry. Hippocampal neurons were treated with rhCNTF in 100 ng·mL-1 concentration and 10 μmol·L-1 condensedAβ. Cell viability affected by rhCNTF and Aβ was determined by CCK-8. The effect of rhCNTF on early apoptosis of hippocampalneurons induced by Aβ was detected by the flow cytometry. Results： Hippocampal neurons were successfully cultured invitro, which can be identified by immunocytochemical method. Cell viability of hippocampal neurons rose with the increasingconcentrations of rhCNTF in an exponential model. The rhCNTF significantly promoted the growth and the cell viability ofhippocampal neurons, whether cells treated with Aβ prior to rhCNTF or with Aβ after rhCNTF （P 〈 0.01）. The rhCNTF decreased theearly apoptosis induced by Aβ. Conclusion： The rhCNTF protected cultured hippocampal neurons from Aβ. The mechanism wasassociated with improving cell growth and reducing cell apoptosis.
出处
《中国药物应用与监测》
CAS
2014年第1期11-14,共4页
Chinese Journal of Drug Application and Monitoring
基金
国家自然科学基金资助项目(81001438)
