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目的探讨亚急性1-溴丙烷(1-BP)吸入染毒对雄性大鼠大脑海马区突触特异性标志物突触素(SYP)、谷氨酸受体2亚基(GluR2)、N-甲基-D-天冬氨酸受体2B亚基(NR2B)蛋白表达的影响。方法将48只无特定病原体级成年雄性Wistar大鼠按体质量随机分为对照组和低、中、高剂量组4组,每组12只。采用动式吸入染毒法,对照组给予新鲜空气,各剂量组分别予质量浓度为1 250、2 500和5 000 mg/m31-BP吸入,每天染毒6 h,每周染毒5 d,连续4周。染毒结束后处死大鼠,取出全脑,分离大脑(含海马组织)、脑干和小脑,分别采用实时荧光定量聚合酶链反应和免疫印迹法检测大鼠海马组织SYP、GluR2、NR2B的mRNA和蛋白相对表达水平。结果高剂量组大鼠于染毒第3周时出现反应迟钝和后肢肌力下降等表现。高剂量组大鼠染毒第1~4周体质量均低于同时间点对照组(P<0.01)。高剂量组大鼠全脑、大脑和脑干质量均低于对照组(P<0.05)。高剂量组大鼠海马组织海马3区和齿状回区可见少量神经元细胞坏死。各组大鼠海马组织SYP、GluR2和NR2B的mRNA相对表达水平分别比较,差异均无统计学意义(P>0.05)。各组大鼠海马组织SYP蛋白相对表达水平比较差异无统计学意义(P>0.05);高剂量组大鼠海马组织GluR2蛋白相对表达水平低于对照组(P<0.05),NR2B蛋白相对表达水平高于对照组(P<0.05)。结论大鼠海马组织GluR2和NR2B蛋白可作为1-BP中枢神经毒性的生物标志物,但其具体的生理意义尚需进一步研究。
Objective To investigate the effects of subacute inhalation of 1-bromopropane (1-BP) on the synaptic signal (SYP), glutamate receptor subunit 2 (GluR2) Methyl-D-aspartate receptor 2B subunit (NR2B) protein expression. Methods Forty-eight adult male Wistar rats without specific pathogen were randomly divided into control group and low, medium and high dose groups, with 12 rats in each group. The rats in the control group were given fresh air. The rats in each dose group were inhaled at 1 250, 2 500 and 5 000 mg / ml respectively. The rats were exposed to 6 hours a day for 5 days a week. For 4 weeks. The rats were sacrificed at the end of the exposure, the whole brain was removed, and the brain (including hippocampus), brain stem and cerebellum were isolated. The mRNA expression of SYP, GluR2 and NR2B in the hippocampus were detected by real-time fluorescence quantitative polymerase chain reaction and immunoblotting And protein relative expression level. Results The rats in high dose group showed unresponsiveness and decreased hindlimb muscle strength at the third week of exposure. The body weight of rats in high dose group from the first week to the fourth week were lower than that of the control group at the same time point (P <0.01). The whole brain, brain and brainstem mass in high dose group were lower than those in control group (P <0.05). A small amount of neuronal cell necrosis was seen in hippocampus 3 and dentate gyrus of hippocampus in high dose group. The relative expression levels of SYP, GluR2 and NR2B in hippocampus of rats in each group were not significantly different (P> 0.05). The relative expression level of SYP in hippocampus of rats in each group had no significant difference (P> 0.05); the relative expression level of GluR2 protein in hippocampus of rats in high dose group was lower than that in control group (P <0.05); the relative expression level of NR2B protein Higher than the control group (P <0.05). Conclusion The GluR2 and NR2B proteins in rat hippocampus may be used as biomarkers of 1-BP central neurotoxicity, but their specific physiological significance needs further study.