摘要
目的:观察小檗碱对小鼠原代肝细胞核因子6(HNF6)mRNA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原代肝细胞,用不同浓度的小檗碱(0,1,3,10,30,100μmol/L)和1 mmol/L二甲双胍干预24 h后,采用MTT比色法观察小檗碱对原代肝细胞增殖的影响:采用RT-PCR方法检测HNF6 mRNA表达,同时检测GK的活性.结果:与阴性对照组相比,1,3,10,30μmol/L的小檗碱均能促进HNF6 mRNA的表达(1.00±0.21,1.11±0.06,1.37±0.10,1.40±0.09 vs 0.68±0.02;P<0.01);当小檗碱为10,30μmol/L时,均可上调GK的活性(0.069±0.082,0.080±0.073 vs 0.009±0.007;P<0.05);二者均在30μmol/L时达到最大值.小檗碱为100μmol/L时HNF6 mRNA的表达、GK的活性与阴性对照组无差异,可能与其抑制肝细胞生长有关.结论:小檗碱改善糖代谢的作用可能与其调节HNF6的表达进而调节GK活性有关.
AIM： To observe the expression of hepatocyte nuclear factor 6 （HNF6） and the activity of glucokinase （GK）, the key enzyme in glucose metabolism, and to investigate the possible mechanism of berberine in treating type Ⅱ diabetes.
METHODS： Mouse primary hepatocytes were isolated by an improved single two-step perfusion method. The murine hepatocytes were incubated with berberine （0, 1, 3, 10, 30 and 100 μmol/L） and 1 mmol/L metformin for 24 hours. mRNA expression for HNF6 was determined by RT-PCR and analyzed by consequent quantification, and the activity of GK was detected by an enzyme kinetics method.
RESULTS： Compared with the negative control group, 1, 3, 10 or 30 μmol/L berberine promoted the expression of HNF6 mRNA （1.00 ± 0.21, 1.11 ± 0.06, 1.37 ± 0.10 or 1.40 ± 0.09 vs 0.68 ± 0.02; P 〈 0.01）. At 10 or 30 μmol/L, berberine upregulated GK activity （0.069 ± 0.082, 0.080 ± 0.073 vs 0.009 ± 0.007; P 〈 0.05）. Both of these reached a maximum at a concentration of 30 μmol/L. At 100 μmol/L berberine, expression of HNF6 mRNA and activity of GK decreased, probably because high-concentration berberine inhibited the growth of primary hepatocytes.
CONCLUSION： It is suggested the effects of berberine in improving glucose metabolism may be associated with its up-regulation of HNF6 mRNA expression and induction of hepatic GK activity.
出处
《世界华人消化杂志》
CAS
北大核心
2007年第36期3842-3846,共5页
World Chinese Journal of Digestology
基金
国家自然科学基金
No 30500685
关键词
小檗碱
肝细胞核因子
原代肝细胞
葡萄糖激酶
逆转录聚合酶链反应
Berberine, Hepatocyte Nuclear Factors
Primary Hepatocytes
Glucokinase
Reverse transcription polymerase chain reaction
