摘要目的 了解富含亮氨酸重复结构的神经细胞黏附分子2(LRRTM2)在氯化锂-毛果芸香碱点燃动物颞叶脑组织中的表达,探讨其与癫痫发生的关系.方法 将氯化锂-毛果芸香碱点燃大鼠56只采用随机数字表法分为6h、24h、72h、7d、14d、30d、60d组,用免疫组织化学(免疫组化)、免疫荧光、蛋白印迹方法,检测LRRTM2在点燃大鼠颞叶脑组织中的表达及其变化规律,并与8只对照组大鼠进行比较.结果 免疫组化和免疫荧光显示,LRRTM2主要在大鼠颞叶皮质神经元中表达.免疫组化平均吸光度(A)值实验组(6h至60d组分别为:0.286 ±0.012、0.227±0.008、0.425±0.015、0.509±0.019、0.579±0.018、0.488±0.018、0.566±0.014)同对照组(0.330 ±0.016)相比差异均有统计学意义(t值分别为:3.965、11.987、9.131、14.121、20.452、12.929、22.786,均P＜0.05);蛋白印迹A比值实验组分别为:0.0354±0.0043、0.0174±0.0026、0.0685±0.0064、0.0957±0.0125、0.1044±0.0103、0.0910±0.0108、0.1012±0.0063,同对照组(0.0471±0.0033)相比差异均有统计学意义(t值分别为:4.354、14.191、5.989、7.541、10.565、7.730、15.316,均P＜0.05).LRRTM2蛋白表达在模型组急性期逐渐降低,24 h最低,72 h后开始升高,至慢性期明显升高,与对照组比较,差异均有统计学意义.结论 LRRTM2在氯化锂-毛果芸香碱点燃大鼠模型颞叶脑组织中静止期、慢性期表达明显增高,推测其在癫痫的发生演变中发挥重要作用.

abstractsObjective Leucine-rich repeat transmembrane neuronal protein 2 (LRRTM2) localizes to excitatory glutamatergic synapses,and triggers the formation of excitatory synapses.This study aims to investigate the expression of LRRTM2 protein in the temporal lobe tissue of SD rats induced by lithiumpilocarpine,and explore its roles in epilepsy.Methods Fifty-six Sprague-Dawley (SD) rats were induced by lithium-pilocarpine and randomly divided into 6h,24h,72h,7d,14d,30d and 60d subgroups.Eight SD rats were treated with normal saline instead of pilocarpine as controls. Expression of LRRTM2 protein was accessed by immunohistochemistry,immunofluorcscence and Western blot analysis. Results LRRTM2 protein mainly expressed in neurons of temporal lobe,gradually decreased in acute phase,and then up-regulated in latent and chronic periods.The immunohistochemistry A values in model rats from 6 h,24h,72h,7d,14d,30d and 60d subgroups were 0.286 ±0.012,0.227 ± 0.008,0.425 ± 0.015,0.509 ±0.019,0.579 ± 0.018,0.488 ± 0.018 and 0.566 ± 0.014,respectively,compared to 0.330 ±0.016 in control group ( t =3.965,11.987,9.131,14.121,20.452,12.929 and 22.786,all P＜0.05). Gray value ratios of LRRTM2/β-actin in different groups of model rats were 0.0354 ± 0.0043,0.0174 ± 0.0026,0.0685 ± 0.0064,0.0957 ± 0.0125,0.1044 ± 0.0103,0.0910 ± 0.0108,and 0.1012 ±0.0063,respectively,which were significant differences from control group (0.0471 ± 0.0033,t=4.354,14.191,5.989,7.541,10.565,7.730and15.316,allP＜0.05).Conclusions LRRTM2 protein gradually increases in the neurons of temporal lobe of SD rats treated by lithium-pilocarpine in silence and chronic phases,which indicates that it may play an important role in cpileptogenesis.