XFM及其颉颃剂对大鼠不同脑JNK3MAPK信号转导通路的影响

发布时间：2018-10-09 21:46

【摘要】：本实验旨在研究小型猪复合麻醉剂(XFM)及其颉颃剂对大鼠不同脑区JNK3 MAPK信号转导通路的影响,探讨XFM及其颉颃剂发挥麻醉和催醒交互作用机制,以便完善对麻醉与催醒机制的认识,为研制、推广新型药物及更好地指导临床实践提供理论依据。将78只SD大鼠随机分为麻醉组(M组)、颉颃剂组(X组)和麻醉剂与颉颃剂交互组(MX组)。M组又分为5个亚组,分别为C1组(对照组)、M1组(注射XFM后翻正反射消失即刻)、M2组(注射XFM后翻正反射消失后1h)、M3组(注射XFM后翻正反射恢复即刻)和M4组(注射XFM后翻正反射恢复后1h);X组又分为3个亚组,分别为C2组(对照组)、X1组(注射小型猪复合麻醉颉颃剂5min)和X2组(注射小型猪复合麻醉颉颃剂1h);MX组又分为5个亚组,分别为C3组(对照组),MX1组(注射XFM,翻正反射消失时立即注射小型猪复合麻醉颉颃剂,翻正反射恢复即刻)、MX2组(翻正反射消失时立即注射XFM复合颉颃剂,翻正反射恢复后1h);MX3组(翻正反射消失后1h,注射XFM复合颉颃剂,翻正反射恢复即刻);MX4组(翻正反射消失后1h,注射XFM复合颉颃剂,翻正反射恢复后1h);各组大鼠到达预订时间点后即刻断头处死,分离大脑皮层、小脑、海马、脑干和丘脑,用实时荧光定量PCR方法检测β-arrestin2基因mRNA的相对转录量。用Western blot的方法检测β-arrestin2、p-JNK和p-ATF-2蛋白的相对表达量。实验结果表明:(1)麻醉剂组,大鼠大脑皮层、小脑中β-arrestin2 mRNA相对表达显著降低,而海马、脑干和丘脑中则显著升高;大脑皮层、小脑和海马中β-arrestin2蛋白、p-JNK蛋白相对表达量均显著降低,而脑干和丘脑中则显著升高;大鼠各脑区p-ATF-2蛋白的相对表达量在小脑和丘脑中显著升高,在海马中显著降低,而大脑皮层与脑干中该蛋白均无显著变化。(2)颉颃剂组,大鼠大脑皮层、小脑中的β-arrestin2 mRNA及其蛋白相对表达量显著升高,而脑干、丘脑中则显著降低,海马中β-arrestin2 mRNA显著降低但其蛋白则显著升高;大脑皮层、小脑和海马中p-JNK蛋白、p-ATF-2蛋白变化趋势与β-arrestin2蛋白一致。(3)麻醉剂与颉颃剂交互作用组,大鼠在早期催醒中大脑皮层、小脑、海马、脑干和丘脑中β-arrestin2 mRNA的相对表达量显著降低,晚期催醒中大脑和小脑β-arrestin2 mRNA的相对表达量显著升高,海马、脑干和丘脑中则显著降低;而β-arrestin2蛋白的相对表达量在大脑皮层、海马中显著降低,在小脑、脑干与丘脑中则显著升高;p-JNK蛋白的相对表达量在大脑皮层、小脑和海马中显著升高,在脑干、丘脑中显著降低;p-ATF-2蛋白的相对表达量在海马显著升高,在丘脑显著降低,但在其他脑区无显著变化。综上所述,XFM可以抑制β-arrestin2蛋白、p-JNK蛋白和p-ATF-2蛋白在大鼠海马中的表达并增强这三种蛋白在大鼠丘脑中的表达,小型猪复合麻醉颉颃剂作用与XFM相反,JNK3 MAPK信号转导通路是XFM及小型猪复合麻醉颉颃剂分别发挥麻醉与催醒作用的重要途径;β-arrestin2蛋白可能是JNK3 MAPK信号转导通路被XFM及小型猪复合麻醉颉颃剂激活的关键位点。小型猪复合麻醉颉颃剂不能完全逆转XFM对JNK3 MAPK信号转导通路的影响,该通路不是XFM产生麻醉作用及小型猪复合麻醉颉颃剂产生催醒作用的唯一途径。
[Abstract]:The purpose of this experiment was to study the effect of XFM and its anti-inflammatory agent on the signal transduction pathway of JNK3 MAPK in different brain regions of rats. It provides a theoretical basis for the promotion of new drugs and better guiding clinical practice. Seventy-eight SD rats were randomly divided into anesthesia group (group M), group (X group) and anesthetic agent group (MX group). The M groups were divided into 5 subgroups, C1 (control group), M1 group (after injection XFM reverse reflex disappeared), M2 group (injection XFM backward positive reflection disappeared 1h), M3 group (injection XFM backward positive reflex recovery immediately) and M4 group (after injection XFM reverse reflex recovery 1h); The X group was divided into three subgroups: group C2 (control group), group X1 (injection minipig compound anesthesia agent for 5min) and X2 group (injection minipig compound anesthesia agent for 1h); MX group was divided into 5 subgroups, C3 (control group) and MX1 group (injection XFM, respectively). Immediate injection of small-sized pig compound anesthesia (anti-inflammatory agent, reverse reflex recovery), MX2 group (immediate injection of XFM composite anti-inflammatory agent after reverse reflex recovery), MX3 group (1h after reverse reflex disappearance), MX3 group (1h after disappearance of flip reflex), and injection of XFM compound anesthetic agent, Revert reflex (immediately); group MX4 (1h after reversal reflex disappearance, injection of XFM compound eye drops, 1h after reverse reflex recovery); end decapitation immediately after arrival of each group of rats, separate cerebral cortex, cerebellum, hippocampus, brain stem and thalamus, The relative amounts of mRNA were detected by real-time fluorescence quantitative PCR. The relative expression of p27-arrestin2, p-JNK and p-ATF-2 protein was detected by Western blot. The results showed that: (1) The relative expression of p27-arrestin2 mRNA in the rat cerebral cortex and the cerebellum was significantly decreased, while in the hippocampus, the brain stem and the thalamus, the relative expression level was significantly decreased in the cerebral cortex, the cerebellum and the hippocampus. The relative expression of p-ATF-2 protein in the brain regions of rats increased significantly in the cerebellum and thalamus, significantly decreased in the hippocampus and no significant changes in the protein in the cerebral cortex and the brain stem. (2) The relative expression of p27-arrestin2 mRNA and its protein in the cerebral cortex and cerebellum of the rat cerebral cortex and the cerebellum increased significantly, while in the brain stem and thalamus, the expression of p27-arrestin2 mRNA in the hippocampus was significantly reduced, but its protein increased significantly; the p-JNK protein in the cerebral cortex, cerebellum and hippocampus, The change trend of p-ATF-2 protein was consistent with that of p27-arrestin2 protein. (3) The relative expression of OPG-arrestin2 mRNA in the cerebral cortex, cerebellum, hippocampus, brain stem and thalamus was significantly decreased in the early awakening of the rats. The relative expression of p-JNK increased significantly in the cerebral cortex and hippocampus, and the relative expression of p-JNK increased significantly in the cerebral cortex, cerebellum and hippocampus, and in the brain stem, There was a significant decrease in the thalamus; the relative expression of p-ATF-2 protein increased significantly in the hippocampus, significantly decreased in thalamus, but there was no significant change in other brain regions. In conclusion, XFM can inhibit the expression of p27-arrestin2 protein, p-JNK protein and p-ATF-2 protein in the hippocampus of rats and enhance the expression of these three proteins in the thalamus of rats. The MAPK signal transduction pathway of JNK3 is an important way for XFM and small pig compound anesthesia to play an important role in anesthesia and awaking. The p38-arrestin2 protein may be the key site of the activation of the JNK3 MAPK signal transduction pathway by XFM and small pig compound anesthesia. The effect of XFM on the signal transduction pathway of JNK3 MAPK can not be completely reversed by a small pig compound anesthesia and anti-inflammatory agent.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S865.1;S857.124

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