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脑源性神经营养因子前体对小鼠海马神经元存活和突起生长的抑制作用
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脑源性神经营养因子前体对小鼠海马神经元存活和突起生长的抑制作用
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Journal Article
脑源性神经营养因子前体对小鼠海马神经元存活和突起生长的抑制作用
2014
Overview
目的探讨脑源性神经营养因子前体（proBDNF）对小鼠海马神经元存活和突起生长的影响,以及p75神经营养素受体（p75NTR）在其中的介导作用。方法取p75NTR转基因18日龄胎鼠（野生型p75NTR＋/＋,基因敲除型p75NTR–/–）海马组织,进行新生海马神经元原代培养。p75NTR＋/＋基因型神经元设对照组、proBDNF（30ng/ml）组和proBDNF（30ng/ml）＋p75/Fc（30μg/ml）组,p75NTR–/–基因型神经元设对照组和proBDNF（30ng/ml）组。采用MTT法检测各组处理24h后新生海马神经元的存活情况,免疫荧光染色观察各组处理72h后的突起长度。结果经MAP2和DAPI双重荧光染色鉴定,成功培养出高纯度新生海马神经元。MTT检测结果显示,在p75NTR＋/＋型海马神经元中,proBDNF组570nm处吸光度值（A570）明显小于对照组（P〈0.05）,加入p75NTR受体竞争性抑制剂p75/Fc后与对照组相比差异无统计学意义（P〉0.05）;在p75NTR–/–型海马神经元中,proBDNF组A570值与对照组差异无统计学意义（P〉0.05）。免疫荧光染色显示,p75NTR＋/＋神经元proBDNF组神经突起长度明显小于对照组（P〈0.05）,proBDNF（30ng/ml）＋p75/Fc（30μg/ml）组与对照组比较差异无统计学意义（P〉0.05）;p75NTR–/–神经元proBDNF组神经突起长度与对照组比较差异无统计学意义（P〉0.05）。结论 proBDNF通过p75NTR受体途径抑制神经元的存活和突起生长,具有毒性作用。
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