CAJ | 학술논문

目的研究小檗碱对胰岛素抵抗模型肝细胞11β-羟基类固醇脱氢酶1(11β-HSD1)mRNA表达的影响.方法应用高浓度胰岛素孵育HepG2细胞24h,建立胰岛素抵抗肝细胞模型.将模型细胞分别置于含不同浓度胰岛素和小檗碱的培养液中培养24h,以葡萄糖氧化酶-过氧化物酶(GOD-POD)法测定培养液中的葡萄糖浓度,计算细胞对葡萄糖的吸收率.应用RT-PCR检测小檗碱作用前后模型细胞11β-HSD1 mRNA的表达.结果以含10-7mol/L胰岛素的培养液孵育HepG2细胞24h后,细胞对葡萄糖的吸收明显降低,表明建模成功.模型细胞经高浓度(10 μmol/L)小檗碱处理后,葡萄糖吸收率明显增加[ (42.53±1.99)％ vs (28.16±1.99)％,t=12.9457,P＜0.01],并且高浓度小檗碱与胰岛素对模型细胞有协同促进葡萄糖吸收的作用.模型细胞11β-HSD1 mRNA 表达显著高于非胰岛素抵抗细胞(相对表达量:4.60±0.96 vs 0.67±0.42,t=4.9476,P＜0.05).经高浓度小檗碱干预24h后,模型细胞11β-HSD1 mRNA表达降低,与非胰岛素抵抗HepG2细胞相比差异无统计学意义(相对表达量:1.12±0.35 vs 0.67 ±0.42,t=1.1394,P＞0.05).低浓度(1μmol/L)小檗碱则作用不明显.结论浓度依赖性地下调11β-HSD1 mRNA表达是小檗碱改善胰岛素抵抗的作用机制之一.
목적 연구소벽감대이도소저항모형간세포11β-간기류고순탈경매1(11β-HSD1)mRNA표체적영향.방법 응용고농도이도소부육HepG2세포24h,건립이도소저항간세포모형.장모형세포분별치우함불동농도이도소화소벽감적배양액중배양24h,이포도당양화매-과양화물매(GOD-POD)법측정배양액중적포도당농도,계산세포대포도당적흡수솔.응용RT-PCR검측소벽감작용전후모형세포11β-HSD1 mRNA적표체.결과 이함10-7mol/L이도소적배양액부육HepG2세포24h후,세포대포도당적흡수명현강저,표명건모성공.모형세포경고농도(10 μmol/L)소벽감처리후,포도당흡수솔명현증가[ (42.53±1.99)％ vs (28.16±1.99)％,t=12.9457,P＜0.01],병차고농도소벽감여이도소대모형세포유협동촉진포도당흡수적작용.모형세포11β-HSD1 mRNA 표체현저고우비이도소저항세포(상대표체량:4.60±0.96 vs 0.67±0.42,t=4.9476,P＜0.05).경고농도소벽감간예24h후,모형세포11β-HSD1 mRNA표체강저,여비이도소저항HepG2세포상비차이무통계학의의(상대표체량:1.12±0.35 vs 0.67 ±0.42,t=1.1394,P＞0.05).저농도(1μmol/L)소벽감칙작용불명현.결론 농도의뢰성지하조11β-HSD1 mRNA표체시소벽감개선이도소저항적작용궤제지일.
Objective To investigate the effect of berberine on glucose absorption and mRNA expression of 11 β-hydroxysteroid dehydrogenase type 1 ( 11β-HSD1 ) in insulin-resistant HepG2 cell model.Methods HepG2 cells were incubated with high-concentration insulin for 24 hours to build insulin-resistant cell model.In order to evaluate the cells for insulin resistance,the cells were cultured with different concentrations of insulin for 24 hours.The insulin-resistant cells were treated with different concentrations of berberine and insulin for 24 hours and the non insulin-resistant cells were used as a control.The glucose concentration in culture medium was detected by the method of glucose oxidase-peroxidase (GOD-POD).According to the glucose concentrations in blank medium and those in the medium of culturing cells 24 hours later,the rate of absorpting glucose by the cells was calculated.The mRNA expression of 11β-HSD1 in the insulin-resistant cells was detected by the reverse transcription polymerase chain reaction (RT-PCR).Results Incubated with 10-7mol/L insulin for 24 h,the insulin-resistant cell model had been built.The rate of glucose absorption of the model cell treated with high concentration berberine ( 10 μmol/L) was significantly improved [(42.53 ±1.99)％ vs (28.16±1.99)％,t =12.9457,P ＜0.01].High-concentration berberine showed a strong synergy with insulin on glucose absorption of the model cells.As the cells became resistant to insulin,the mRNA expression of 11β-HSD1 increased significantly compared to non insulin-resistant cells( relative expression quantity was (4.60 ±0.96 vs 0.67 ±0.42,t =4.9476,P ＜0.05 ).While the mRNA expression of 11β-HSD1 reduced in the insulin-resistant cells after treated with high-concentration berberine,and the relative expression quantity was not significantly different with non insulin-resistant cells ( 1.12 ±0.35 vs 0.67 ±0.42,P ＞0.05).However,low-concentration ( 1 μmol/L) of berberine had not the same role.Conclusions It is concluded that one of the acting mechanism of berberine improving the insulin sensitivity may be that the mRNA expression of 11β-HSD1 is downregulated in the insulin-resist-ant liver cell model depending on concentration.