摘要
目的 应用针对靶向水通道蛋白4（aquaporin 4，AQP-4）的小RNA（siRNA）干扰技术优化亚低温减轻颅脑创伤（traumatic brain injury，TBI）后脑水肿程度的治疗效果。方法构建沉默AQP-4mRNA表达的siRNA质粒；液压打击法建立大鼠TBI模型，分TBI对照组、AQP-4siRNA治疗组、亚低温治疗组、AQP-4 siRNA及亚低温联合治疗组；提取第1、3、5、7天脑组织总RNA和总蛋白，RT—PCR和Westernblot方法检测AQP-4的mRNA和蛋白表达水平；干／湿比重法和Evans蓝测定法观察大鼠TBI后不同时相脑组织含水量和血脑屏障通透性改变；实验动物予以神经功能缺陷综合评分。结果亚低温在减轻TBI后脑水肿程度方面优于AQP-4siRNA，但siRNA技术在沉默AQP-4表达方面强于亚低温，联合应用AQP-4siRNA和亚低温在TBI后降低脑水肿程度方面获得最佳治疗效果。结论靶向AQP-4的SiRNA干扰技术可优化亚低温在TBI后降低脑水肿方面的治疗效果。
Objective To optimize the treatment effect of siRNA targeting aquaporin 4 with hypotherulia on cerebral edema following traumatic brain injury. Methods To construct siRNA targeting aquaporin 4, and establish the TBI models in rats. The animals were divided into four groups： TBI group, AQP-4 siRNA group, hypotherulia group, and combined group. To extract the total mRNA and proteins on the 1^st, 3^rd, 5^th, and 7^th day for RT-PCR and western blot analysis. The changes of brain water content and permeability of blood-brain barrier were measured by the methods of wet and dry weight and Evans blue fluorometry. All the rats were given a mark for their nerve function on the 7^th day. Results Hypothermia treatment can obtain better effect than AQP-4 siRNA in lessening brain edema level. Meanwhile, siRNA treatment can obtain better effect than hypothermia group in silencing AQP-4 mRNA and protein expression. When combining hypothermia treatment with AQP-4 siRNA, we can obtain an optimized result than the single application of above mentioned two groups. Conclusion The application of siRNA targeting aquaporin 4 may optimize the treatment effect of hypothermia on traumatic brain injury rats.
出处
《中华神经外科杂志》
CSCD
北大核心
2009年第2期178-181,共4页
Chinese Journal of Neurosurgery
基金
天津市科委攻关课题基金（06YFSZSF01200）
