异氟醚后处理的脑保护作用及其机制的研究

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1、《中国图书资料分类法》单位代码：10660分类号：R614学号：S100376贵阳医学院2013届临床科学硕士学位专业材料异氟醚后处理的脑保护作用及其机制的研究研究生：赵倩导师:章放香教授年级：2010级专业：麻醉学2013年05月10日目录摘要································································1前言································································3材料与方法·············

2、··············································5结果································································13讨论································································21结论································································25参考文献··················

3、···········································26英文摘要·····························································31致谢································································33略缩词表·····························································34论文原创性声明···················

4、··································35附：综述····························································36贵阳医学院2013届研究生论文异氟醚后处理的脑保护作用及其机制的研究专业麻醉学研究生赵倩导师章放香教授摘要目的通过观察异氟醚后处理对SD大鼠脑缺血再灌注后海马CA1区神经细胞凋亡及凋亡蛋白Caspase-3表达的影响，探讨异氟醚后处理的脑保护效应、后处理时间窗以及JNK信号通道在异氟醚后处理诱导脑保护中的作用。方法健康成年SD大鼠40只，随

5、机分为5组（n=8），A组：正常对照组，仅手术操作，不进行缺血再灌注；B组：缺血再灌注损伤组，电凝双侧椎动脉，24小时后分离双侧颈总动脉并夹闭15分钟后开放；C组：异氟醚后处理Ⅰ组，再灌注即刻吸入异氟醚；D组：异氟醚后处理Ⅱ组，再灌注10min吸入异氟醚；E组：异氟醚后处理Ⅲ组，再灌注20min吸入异氟醚。异氟醚后处理吸入浓度为1.5%，各处理组分别于再灌注时间点吸入异氟醚30min。再灌注72h后取脑组织常规HE染色观察大鼠海马神经中神经系统损伤，同时应用免疫组织化学染色检测Caspase-3在SD大鼠海马神经细胞中表达，TUNEL法检测

6、大鼠海马中神经细胞的病理学变化，免疫印迹法（Westernblotting）检测大鼠海马组织p-JNK蛋白表达量。结果1.HE染色：A组脑组织切片未见明显海马组织损伤；B组海马组织损伤明显,显示颗粒层细胞明显水肿和空泡变性,神经细胞坏死，血管明显充血并出血；与B组相比较，D、C组损伤的程度依次减轻(P<0.05)，E组损伤程度与B组相似(P>0.05)。2.Caspase-3免疫组化检测：A组未检测到明显的Caspase-3阳性神经元表达；B组Caspase-3的阳性表达明显增高;与B组相比较，D、C组Caspase-3的阳性表达依次降低(

7、P<0.05)，E组阳性表达与B组相似（P>0.05）。3.TUNEL法细胞凋亡检测：A组未见明显的神经细胞凋亡，B组神经细胞凋亡阳性表达显著增高；与B组相比较，D、C组细胞凋亡阳性表达率依次降低(P<0.05)，E组的细胞凋亡阳性表达率与B组相似(P>0.05)。4．免疫印迹法p-JNK蛋白表达检测：与A组比较，B组p-JNK蛋白阳性表达量显著增高；与B组相比较，C、D组p-JNK蛋白阳性表达量明显降低（P<0.05）,C、1贵阳医学院2013届研究生论文D组p-JNK蛋白表达量无明显差异（P>0.05），E组p-JNK蛋白阳性表达量与B

8、组无明显差异（P>0.05）。结论1.异氟醚后处理通过抑制Caspase-3的表达，抑制神经细胞凋亡，减轻脑缺血再灌注损伤，产生脑保护效应。2.异氟醚后处理具有一定的时效性，脑损