单纯性脑震荡对大鼠海马iNOS表达影响的实验研究
李恒希,吴德野,赖江华,康丽,吴海鹰,于建云,李坪
中国临床解剖学杂志 ›› 2016, Vol. 34 ›› Issue (4) : 402-406.
单纯性脑震荡对大鼠海马iNOS表达影响的实验研究
Experimental research on the expression of iNOS in the hippocampus of rats after pure cerebral concussion
目的 观察单纯性脑震荡( pure cerebral concussion, PCC )大鼠海马脑区诱导性一氧化氮合酶( inducible nitric oxide synthase, iNOS )的表达变化，探讨iNOS表达变化规律及其与PCC之间的病理联系。 方法 采用单摆闭合式脑损伤打击装置制造PCC大鼠模型，造模后随机分为3 h、12 h、1 d、2 d、3 d、7 d 六个损伤组( n=5 )，另设正常对照组( n=5 )。采用iNOS单克隆抗体(鼠)进行免疫组织化学SP法染色和Western Blot实验，检测PCC组和正常组大鼠海马( CA1~4及上、下齿状回 ) iNOS表达的定位及定量情况。 结果 正常生理状态下，海马iNOS阳性细胞染色较浅、轮廓不清，且iNOS蛋白表达较弱，致伤后3 h组iNOS阳性表达开始增强，在3 d组出现表达高峰，iNOS蛋白表达高峰出现在2 d组，与正常组比较均有显著性差异( P<0.05 )，随后均出现下降，至7 d均仍高于正常组。 结论 PCC损伤早期海马脑区出现iNOS表达增强，提示iNOS是参与PCC继发性病理损伤的主要炎症因子之一。
Objective To investigate the expression change pattern and its pathological link between iNOS and PCC though observing expression changes of iNOS (inducible nitric oxide synthase, iNOS) between hippocampus CA1-4 region and upper and lower dentate gyrus regions in the pure cerebral concussion (PCC) model rat. Method PCC model was established insober rats by using metal pendulum closed brain injury blow device, andthen the rats were randomly divided into 3h, 12h, 1d, 2d, 3d, 7d six injury groups (n=5) and a normal control group (n=5). Location and quantitation of iNOS in hippocampal CA1-4 and superior and inferior dentate gyrus between the PCC groups and the normal group were detected immunohistochemical staining and Western Blot. Result Under normal physiological conditions, iNOS positive cells in hippocampus were few, poorly defined and the protein expression was weak. However, 3h after injury, iNOS and the positive protein expression levels in hippocampus increased and reached the peak on 3d and 2d, respectively. There was a significant difference with the normal group (P<0.05), after 2d, the expression of protein was decreased and the positive expression was also decreased after 3 d, but the expression was still higher than that of normal group until 7 d. Conclusion Expression of iNOS in the hippocampus increased at the early stage of PCC damage, suggesting that the inflammatory factor plays an important role in the pathological changes of PCC.
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国家自然科学基金资助项目(31100769，30560041)；云南省自然科学基金 (2008CD052)
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