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旨在从细胞学实验及整体动物水平探讨几丁寡糖(NACOS)对机体脂代谢紊乱的抑制作用及其潜在的分子机制。在细胞学实验中,HepG2细胞被分为4组,即对照组、棕榈酸(Palmitic acid,PA)组、几丁寡糖(NACOS)组、NACOS+PA组。在体内实验中,将雄性C57BL/6小鼠随机分为4组(n=5),即正常对照(NCD)组、高脂饮食(HFD)组、NACOS组、NACOS+HFD组,实验共20周。主要检测方法如下:采用油红O染色检测细胞脂质沉积,RT-PCR方法检测脂代谢调控分子及炎症因子的转录表达水平,Western blotting方法检测MAPKs及PI3K/Akt通路中相关蛋白激酶的蛋白磷酸化水平。细胞学实验表明,NACOS对HepG2没有明显的细胞毒性作用,并能显著降低细胞内脂滴颗粒的沉积,下调肝细胞及小鼠肝脏组织中脂代谢相关调控因子(PGC1α、Cox5b及Mcad)及炎症因子IL-1β的转录表达水平(P<0.05或0.01),抑制肝细胞及肝脏组织中p38、ERK1/2及Akt蛋白激酶的激活(P<0.05或0.01)。基于上述研究,NACOS可抑制肝脏线粒体脂肪酸氧化和脂质从头合成途径,阻断炎症反应的发生,从而预防脂代谢紊乱的发生。
The aim of this study is to investigate the inhibitory effect of NACOS on the lipid metabolism disorder and its potential molecular mechanisms from cytology experiments and overall animal studies. In cytology experiments, HepG2 cells were divided into 4 groups, namely control group, palmitic acid (PA) group, chitin oligosaccharides (NACOS) group, NACOS + PA group. In vivo experiments, male C57BL / 6 mice were randomly divided into four groups (n = 5), ie, normal control group (NCD), high fat diet group (HFD), NACOS group, NACOS + HFD group week. The main detection methods were as follows: Lipid deposition was detected by oil red O staining, the transcriptional expression of lipid metabolism regulators and inflammatory cytokines were detected by RT-PCR, the protein phosphatases of MAPKs and PI3K / Akt pathway were detected by Western blotting Level Cytology experiments showed that NACOS had no obvious cytotoxic effect on HepG2, and could significantly reduce the deposition of intracellular lipid droplets, downregulate the lipid metabolism related regulators (PGC1α, Cox5b and Mcad) and inflammation in liver cells and mouse liver tissues (P <0.05 or 0.01), and inhibited the activation of p38, ERK1 / 2 and Akt protein kinases (P <0.05 or 0.01) in hepatocytes and liver tissues. Based on the above studies, NACOS can inhibit the liver mitochondrial fatty acid oxidation and lipid de novo synthesis pathway, blocking the occurrence of inflammatory reactions, thus preventing the occurrence of lipid metabolism disorder.