目的：观察青钱柳多糖对肝细胞胰岛素信号传导通路关键靶点基因、蛋白表达的影响。方法：以H4IIE大鼠肝细胞为模型，培养72h后，采用Neutral Red法检测青钱柳多糖（50、100、200、400μg·mL^-1）对细胞活性的影响；根据细胞活性检测结果分为正常对照组（Control组）、胰岛素组（Insulin组）、青钱柳多糖低剂量组（LCP组）和高剂量组（HCP组）；药物干预2h后RT-PCR检测肝细胞相应基因表达，药物干预30min后，利用Western blot法检测肝细胞相应蛋白磷酸化水平。结果：与Control组比较，100、200、400μg·mL^-1青钱柳多糖干预后肝细胞活性显著升高（P<0.01）；干预2h 后，LCP组InsR、IRS-2基因表达显著升高（P<0.05），Insulin组和HCP组InsR、IRS-2基因表达极显著升高（P<0.01）；干预30min后，Insulin组、HCP组InsR β、IRS-2、Akt蛋白磷酸化水平显著升高（P<0.01）。结论：青钱柳多糖具有增加肝细胞活性作用，其上调肝细胞胰岛素信号通路关键靶点InsR、IRS-2基因表达，以及InsR β、IRS-2、Akt蛋白磷酸化水平可能是其发挥降糖作用的机制。

This study aimed to explore the impacts of Cyclocarya paliurus polysaccharide(CP)on insulin signal pathway in liver cells. H4IIE liver cells of rat that cultivated for 72h were made up for the model group.H4IIE cells at 70%-80% confluence were exposed to various concentrations of CP,including 50,100,200 and 400μg·mL^-1,for measuring cell viability using Neutral Red.Based on the results of cell viability,H4IIE cells were divided into the control group,the insulin group(100nmol·L^-1),the low dose CP group(the LCP group,200μg·mL^-1)and the high dose CP group (the HCP group,400μg·mL^-1).After the cultivation of cells for 2h,mRNA levels were measured by real-time RT-PCR,while phosphorylation levels of target proteins were detected by western blot after the treatment for 30min.It was found that the cell viability indexes in the cells administered by 100,200 and 400μg·mL^-1 CP increased significantly compared with the control group(P<0.01).After the administration for 2h,InsR and IRS-2 mRNA expressions in the insulin group,the LCP group and the HCP group increased(P<0.05 or P<0.01).After the administration for 30min,phosphorylation levels of InsRβ,IRS-2 and Akt in the insulin group were higher than those in the HCP group(P<0.01).In conclusion,CP probably increased the cell viability of H4IIE cells,and improved the blood glucose index through enhancing the mRNA expressions of InsR and IRS-2 and the phosphorylation levels of I nsRβ,IRS-2 and Akt in the liver.

北京市教育委员会科学研究与研究生培养共建项目-科研项目：中药干预胰岛素抵抗技术平台建设与方药筛选研究，负责人：刘铜华。