小檗碱对肝细胞核因子4α表达及葡萄糖激酶活性的影响  

Effects of berberine on expression of hepatocyte nuclear factor 4α and glucokinase activity in mouse primary hepatocytes

作　　者：闫忠卿[1] 冷三华[1] 陆付耳[1] 陆小红 董慧[1] 高志强[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院中西医结合研究所,湖北武汉430030 [2]湖北天茂集团制剂厂,湖北荆门448000

出　　处：《中国中药杂志》2008年第18期2105-2109,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30500685)

摘　　要：目的:观察小檗碱对小鼠原代肝细胞核因子4α(HNF4α)表达及葡萄糖代谢关键酶葡萄糖激酶(GK)活性的影响,探讨小檗碱治疗2型糖尿病的分子机制。方法:采用改良两步灌流法培养小鼠原代肝细胞,用不同浓度的小檗碱(0,1,3,10,30,100μmol.L-1)和1 mmol.L-1二甲双胍干预24 h后,采用RT-PCR方法检测HNF4αmRNA表达;采用Western-blot方法检测HNF4α蛋白的表达,同时检测GK的活性。结果:与阴性对照组相比,小檗碱在10,30,100μmol.L-1时能够明显促进HNF4αmRNA及蛋白的表达,二者同时在30μmol.L-1时达到最大值(HNF4αmRNA相对吸光度为0.48±0.20,蛋白相对吸光度为3.47±0.86,P<0.01);在10,30μmol.L-1时明显上调GK活性,同时在30μmol.L-1时达到最大值(0.080±0.073,P<0.05);而二甲双胍对HNF4αmRNA、蛋白的表达及对GK活性的影响则与阴性对照组无明显差异。结论:小檗碱改善糖代谢的作用可能与其调节肝细胞核因子4α的表达进而调节GK活性有关。Objective： To observe the expression of hepatocyte nuclear factor 4α （HNF4α） and the activity of key enzyme glucokinase （GK） in glucose metabolism, and further to investigate the possible mechanism of berberine in treating type 2 diabetes. Method： Mouse primary hepatocytes were isolated by an improved single two-step perfusion method. The murine hepatocytes were cultured and incubated with berberine （0, 1, 3, 10, 30, 100 μmol·L-1） and 1 mmol·L-1 metformin for 24 h respectively. The mRNA expression of HNF4α were quantified by RT-PCR and the protein expression of HNF4α were quantified by Western-blot. And the activity of GK were detected with enzyme kinetics method. Result： As compared with the negative control group, at a certain concentration range, the expression of HNF4α mRNA and protein and the activity of GK were promoted by berberine. Both of them reached the top at the concentration of 30μmol· L-1 （P 〈0. 01 ）. But the metformin made no difference with the negative control group on the expression of HNF4α and the activity of GK. Conclusion： It is suggested that the effects of berberine on improving glucose metabolism can be mechanically associated with its up-regulating the HNF4α expression and inducing the activity of hepatic glucokinase.

关 键 词：小檗碱 肝细胞核因子 葡萄糖激酶 原代肝细胞 

分 类 号：R285.5[医药卫生—中药学]