大鼠侧脑室注射Nesfatin-1对糖脂代谢的影响及其与下丘脑AMPK表达的关系

摘要

目的 探讨侧脑室注射摄食抑制因子Nesfatin-1对糖脂代谢的影响及其与下丘脑AMPK表达的关系.方法 借助脑立体定位技术将Nesfatin-1(1-82)重组蛋白注射至大鼠侧脑室(实验组),设注射等体积人工脑脊液的假手术组和未施加任何干扰因素的对照组,大鼠继续饲养4w后,全自动生化分析仪检测血糖血脂水平变化,RT-PCR法检测大鼠下丘脑组织AMPK mRNA表达,Western blotting法检测大鼠下丘脑组织AMPKα及磷酸化AMPKα(pAMPKα)的表达.结果 (1)生化检测实验结果显示,实验组大鼠FPG和TG与对照组比较均显著下降(P<0.05),但TC、HDL和LDL水平未见明显变化(P>0.05);(2)RT-PCR及Western blotting实验结果显示,与对照组比较,实验组的AMPK mRNA及蛋白表达水平均明显降低(P<0.05),假手术组的AMPK mRNA及蛋白表达水平与对照组比较均无显著性差异(P>0.05).结论 Nesfatin-1可能在糖脂代谢神经调节过程中具有重要作用,其机制可能与下丘脑AMPK表达水平有关.%Objective To investigate the effect of injecting the feeding inhibitory factor Nesfatin-1 into Lateral ven-tricle on glucolipid metabolism and its connection with AMPK expression of hypothalamus in rats .Methods With the help of the brain stereotaxic instrument,Nesfatin-1(1-82) recombinant protein (experimental group) and equal artificial cerebro-spinal fluid ( sham-operated group ) were injected into the lateral ventricle of rats ,the control group did not exert any inter-ference.4 weeks later,testing the level of blood glucolipid by automatic biochemical analyzer and detecting the expression of AMPK mRNA,AMPKαand pAMPKαin hypothalamus through RT-PCR and Western blotting.Results (1)According to the results of biochemical tests ,compared to the control group ,the level of FPG and TG in experimental group were signifi-cantly decreased (P<0.05),while the level of TC,HDL and LDL were not significant change (P>0.05);(2)According to the results of RT-PCR and Western blotting ,compared to control group ,the expression of AMPK mRNA and protein level in experimental group were significantly lower (P<0.05) and shown no significant difference (P>0.05) with sham-oper-ated group.Conclusion Nesfatin-1 may play an important role in the process of neuroregulation on glucolipid metabolism , the mechanism may be related to the AMPK expression level of hypothalamus .