β-1,4-半乳糖基转移酶Ⅴ在创伤性脑损伤中的表达

Expression of β-1,4-GalTⅤ in traumatic brain injury

目的:探讨创伤性脑损伤(traumatic brain injury,TBI)后,大鼠脑组织中β-1,4-半乳糖基转移酶Ⅴ(β-1,4-galactosyltransferaseⅤ,β-1,4-Gal TⅤ)的表达及作用机制。方法:SD大鼠30只,随机分成假手术组、TBI 1 d、3 d、5 d、7 d组(每组6只)。采用实时定量聚合酶链式反应(real time quantitative polymerase chain reaction,q RT-PCR)检测β-1,4-Gal TⅤm RNA的表达变化,Western Blot检测β-1,4-Gal TⅤ的蛋白表达变化,免疫荧光双标观察β-1,4-Gal T V与神经元标志物Neu N、星形胶质细胞标志物GFAP的共定位情况。结果:TBI后β-1,4-Gal TⅤm RNA和蛋白表达水平一致,与假手术组相比,TBI后表达水平均明显升高,并于损伤后3 d达到高峰。免疫荧光双标检测显示,β-1,4-Gal TⅤ与Neu N存在明显的共定位,与GFAP存在部分共定位。结论:β-1,4-Gal TⅤ在TBI后神经元和星形胶质细胞中均发挥一定的作用,以神经元为主。

Objective： To explore the expression and role mechanism of β-1,4-galactosyltransferase Ⅴ（β-1,4-Gal TⅤ） in rat brain after traumatic brain injury（TBI）. Methods： SD rats（n=30） were divided randomly into sham, 1 d, 3 d, 5 d, 7 d postTBI group（n=6 per group）. Real time quantitative polymerase chain reaction（q RT-PCR） was performed to detect the expression change of β-1,4-Gal T Ⅴ m RNA. Western Blot was used to investigate the expression change of β-1,4-Gal T Ⅴ protein.Double-labeling immunofluorescence was used to detect the co-localization of β-1,4-Gal TⅤ with Neu N-marker of neuron or GFAP-marker of astrocyte. Results： The results of q RT-PCR are similar to that of Western Blot, which showed that β-1,4-Gal T Ⅴ m RNA increased markedly after TBI compared with sham group and peaked at 3 d post-TBI. Double-labeling immunofluorescence revealed that there was obvious co-localization of Neu N and β-1,4-Gal TⅤ. GFAP co-located with β-1,4-Gal TⅤpartly. Conclusion： β-1,4-Gal TⅤplays a vital role in neuron and astroctye after TBI, mainly in neuron.