青钱柳多糖对H4IIE肝细胞胰岛素信号传递的影响  

Impacts of Cyclocarya Paliurus Polysaccharide on Insulin Signaling Pathway in H4IIE Liver Cells

作　　者：许光远[1] 孙文[2] 郭璇[1] 候丹[3] 吴丽丽[2] 张岩[1] 张茁[3] 张露[1] 秦灵灵[4] 李博[5] 李伟笠 刘铜华[7] 

机构地区：[1]北京中医药大学附属东方医院,北京100078 [2]北京中医药大学中医养生学研究所,北京100029 [3]陕西中医药大学第一临床医学院,咸阳712046 [4]北京中医药大学科技处,北京100029 [5]北京中医药大学中药学院,北京100029 [6]北京中医药大学东直门医院,北京100700 [7]北京中医药大学研究生院,北京100078

出　　处：《世界科学技术-中医药现代化》2016年第7期1144-1148,共5页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：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

摘　　要：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72 h后,采用Neutral Red法检测青钱柳多糖(50、100、200、400μg·m L-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2 h后RT-PCR检测肝细胞相应基因表达,药物干预30 min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·m L-1青钱柳多糖干预后肝细胞活性显著升高(P<0.01);干预2 h后,LCP组Ins R、IRS-2基因表达显著升高(P<0.05),Insulin组和HCP组Ins R、IRS-2基因表达极显著升高(P<0.01);干预30 min后,Insulin组、HCP组Ins Rβ、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Ins R、IRS-2基因表达,以及Ins Rβ、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.H4 IIE liver cells of rat that cultivated for 72 h were made up for the model group.H4 IIE cells at 70%-80% confluence were exposed to various concentrations of CP,including 50,100,200 and 400 μg·m L-1,for measuring cell viability using Neutral Red.Based on the results of cell viability,H4 IIE cells were divided into the control group,the insulin group（100 nmol·L-1）,the low dose CP group（the LCP group,200 μg·m L-1） and the high dose CP group（the HCP group,400 μg·m L-1）.After the cultivation of cells for 2 h,m RNA levels were measured by real-time RT-PCR,while phosphorylation levels of target proteins were detected by western blot after the treatment for 30 min.It was found that the cell viability indexes in the cells administered by 100,200 and 400 μg·m L-1 CP increased significantly compared with the control group（P〈0.01）.After the administration for 2 h,Ins R and IRS-2 m RNA expressions in the insulin group,the LCP group and the HCP group increased（P〈0.05 or P〈0.01）.After the administration for 30 min,phosphorylation levels of Ins Rβ,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 H4 IIE cells,and improved the blood glucose index through enhancing the m RNA expressions of Ins R and IRS-2 and the phosphorylation levels of Ins Rβ,IRS-2 and Akt in the liver.

关 键 词：青钱柳多糖 H4IIE肝细胞 胰岛素信号传导通路 

分 类 号：R587.1[医药卫生—内分泌]