摘 要: 目的 探讨钙蛋白酶拮抗剂Calpeptin对锰干扰突触体囊泡融合的保护作用。方法 将24只小鼠随机分成4组,每组6只:对照组(腹腔注射0.9%氯化钠),低、高剂量染锰组(腹腔注射25 μmol/ kg 、100μmol/ kg氯化锰),Calpeptin预处理组(皮下注射Calpeptin 100 μg/kg,30min后腹腔注射100 μmol/ kg氯化锰),注射容量为2 ml/ kg,每周5 次, 共4周。处死小鼠后分离基底核,制备突触体,测量小鼠基底核内锰含量,神经细胞内钙离子浓度和钙蛋白活力,突触体SNARE复合物及其相关蛋白的变化和囊泡融合情况。结果 与对照组比较,高剂量染锰组小鼠基底核锰含量,神经细胞内钙离子浓度和钙蛋白酶活力显著增加;SANP25蛋白表达下降,并出现裂解碎片,SNARE复合物形成减少,FM1-43荧光强度的下降;Calpeptin预处理可以明显抑制神经细胞内钙蛋白酶活力,缓解钙蛋白酶对SNAP25的裂解,100 kDa SNARE复合物也明显增多,而且SNARE复合物介导的突触囊泡融合有所上升。结论 钙蛋白酶拮抗剂Calpeptin可以对锰干扰突触体囊泡融合起到有效的保护作用。 |
关键词: 氯化锰 钙蛋白酶拮抗剂 突触体 SNARE复合物 神经递质囊泡 |
中图分类号: R994.6
文献标识码:
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基金项目: 国家自然科学基金资助(编号:81372942);辽宁省高等学校杰出青年学者成长计划(编号:LJQ2014089) |
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Protective effect of calpain antagonist Calpeptin on disturbance of manganese to synaptosomal vesicles fusion in mouse basal nuclei |
XU Bin, WANG Can, HOU Xiao-yu , ZHOU Ying-hao, DENG Yu, LIU Wei, XU Zhao-fa
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School of Public Health,China Medical University,Shenyang 110122,China
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Abstract: Objective To study the protective effect of calpain antagonist Calpeptin (CP) on disturbance of manganese to synaptosomal vesicles fusion in mouse basal nuclei. Methods Twenty-four mice were randomly divided into four groups: first group was control group the mice were given 0.9% NaCl solution intraperitoneally (i.p.); the mice in second and third group were given 25 and 100 μmol/kg Mncl2 i.p, respectively; the mice of fourth group were subcutaneously injected (s.c) with 100 μg/kg of CP and i.p. injected with 100 μmol/kg of Mncl2 30 minutes later after administration of CP. The injection capacity all were 2 ml/kg, five times a week, for 4 weeks. After the last injection, mice were sacrificed, the synaptosomes of basal nuclei were isolated and extracted for the detection of Mn level , calcium content and calpain activity in basal nuclei .Meanwhile, the changes of synaptosomal SNARE complexes and its related proteins, the vesicle fusion etc. were measured as well. Results Compared with control group, there were significant increase in basal nuclei Mn level , intracellular calcium content and calpain activity, while the SNARE complex formation, SANP25 protein expression and FM1-43 fluorescence intensity were all decreased in 100 μmol/kg Mncl2 group, SNAP-25 protein fragments could also be seen. The pretreatment of 100 μg/kg CP could significantly inhibit calpain activity, alleviate the SNAP25 cleavage by calpain, and 100 kDa SNARE complexes and vesicle fusion also increased significantly. Conclusion Calpain antagonist Calpeptin shows some protective effect on the disturbance induced by manganese to synaptomal vesicles fusion in mouse basal nuclei. |
Keywords: manganese calpain antagonist synaptosomes SNARE complexes neurotransmitter vesicles |