中国临床解剖学杂志 ›› 2017, Vol. 35 ›› Issue (1): 52-56.doi: 10.13418/j.issn.1001-165x.2017.01.011

• 实验研究 • 上一篇    下一篇

大鼠急性甲醇中毒大脑皮质单核细胞趋化蛋白-1蛋白的表达

李璠1, 李红娥2, 韩宏2, 曹小青2, 赵敏2, 李娟娟2, 袁云2, 吴春云2   

  1. 1. 昆明医科大学科研实验中心;    2. 昆明医科大学人体解剖学与组织胚胎学系,  昆明   650500
  • 收稿日期:2016-10-10 出版日期:2017-01-25 发布日期:2017-02-22
  • 通讯作者: 吴春云,教授,博士生导师,Tel:13888797152,E-mail:wuchunyunkm@163.com
  • 作者简介:李璠(1984-),女,云南临沧市人,硕士,助理实验师,主要从事基础医学及分子生物学实验技术工作,Tel:13759494469, E-mail:282067107@qq.com
  • 基金资助:

    国家自然科学基金项目(31260254);云南省应用基础研究计划重点项目(2015FA020)

Expression of monocyte chemoattractant protein-1 protein in cerebral cortex of rats after acute methanol poisoning

LI Fan1,LI Hong-e2,HAN Hong2,CAO Xiao-qing2,ZHAO Min2,LI Juan-juan2,YUAN Yun2,WU Chun-yun2   

  1. 1. Department of Experiment Center for Medical Science Research,Kunming Medical University ,Kunming 650500,China; 2. Department of Anatomy/Histology and Embryology, Kunming Medical University,Kunming 650500,China
  • Received:2016-10-10 Online:2017-01-25 Published:2017-02-22

摘要:

目的 观察大鼠急性甲醇中毒后单核细胞趋化蛋白1(MCP-1)在大脑皮质的表达变化。 方法 应用通有混合气体(N2O/O2)的密闭有机玻璃箱并灌胃相应剂量甲醇溶液复制急性高剂量和低剂量甲醇中毒大鼠模型,运用Western Blot和免疫组化染色检测不同时间点大脑皮质中MCP-1的表达变化。 结果 Western Blot显示急性甲醇中毒后低剂量组和高剂量组MCP-1蛋白表达在12 h明显升高(P<0.01),24 h达高峰,然后逐渐下降,3 d仍高于对照组(P<0.01)。高剂量组MCP-1蛋白表达在12 h和24 h高于低剂量组(P<0.05),3 d 组MCP-1的表达无显著差异。免疫组化染色显示对照组大鼠大脑未见明显MCP-1阳性表达,急性甲醇中毒后,高剂量组24 h MCP-1阳性表达数增加,3 d达高峰,和对照组相比有显著差异(P<0.01),阳性表达多集中在梨状皮质,1周阳性反应减少,但与对照组相比仍有显著差异(P<0.01)。  结论 大鼠急性甲醇中毒后,脑组织MCP-1蛋白表达显著增高并持续一定时间,可能参与了脑组织病理变化过程,程度与甲醇中毒剂量有关。

关键词: 单核细胞趋化蛋白1, 急性甲醇中毒, 大鼠

Abstract:

Objective This study was aimed to investigate the expression of monocyte chemoattractant protein-1 (MCP-1) proteinin rats after acute methanol poisoning. Methods Animal models of acute methanol poisoning in rats were duplicated using a plexiglas chamber exposed to a mixture of N2O/O2. Perfusion fixation, paraffin embedding, and sectioning were used, and positive cells expression was observed. Extract protein from brain tissue and Western blot were use to detected the expression level of MCP-1 protein. Results Results of Western blot: The expression level of MCP-1 were significantly enhanced with over time following acute poisoning, reaching the maximum at 24h; Expression in high-dose group was higher than low-dose group at 12h and 24h (P<0.01). There were no significant difference between the low-dose group and high-dose group at 3d (P>0.05). Results of immunohistochemistry experiment: MCP-1 positive cells reaction could be found in piriform cortex in the high-dose group, whereas the saline group showed no brown-yellow granules scattered. The expression level of MCP-1 were significantly increasedat 24 h (P<0.01), reached the maximum at 3d (P<0.01),and then decreased at 1week, but still had significant difference compared with the saline control group (P<0.01). Conclusion The expression level of MCP-1 proteinare significantly enhanced and last a period of time following acute methanol poisoning; the degree of poisoning and the dose administered is apparently related, and MCP-1 might participate in brain pathological process.

Key words: Monocyte chemoattractant protein-1, Methanol poisoning, Rats