论文部分内容阅读
目的:观察小檗碱对高糖诱导的3T3-L1脂肪细胞胰岛素抵抗模型核因子NF-κB p65表达及转位的影响,探讨小檗碱改善胰岛素抵抗的分子生物学机制。方法:以25mmol·L-1葡萄糖加0.6nmol·L-1胰岛素诱导3T3-L1脂肪细胞产生胰岛素抵抗,以小檗碱进行干预,同时阿司匹林作为阳性对照,以2-脱氧-[3H]-D-葡萄糖摄入法推算葡萄糖的转运率,用Western blot检测3T3-L1脂肪细胞总NF-κB p65蛋白及核NF-κBp65蛋白的表达,激光扫描共聚焦(CLSM)对NF-κB p65蛋白进行定位显示。结果:25mmol·L-1葡萄糖加0.6nmol·L-1胰岛素作用18h后使3T3-L1脂肪细胞胰岛素刺激的葡萄糖转运率抑制60%,Western blot显示核NF-κB p65蛋白表达明显增加,CLSM显示NF-κB p65核转位增加;同时加入小檗碱或阿司匹林则可逆转上述效应。但高糖、小檗碱、阿司匹林对3T3-L1脂肪细胞总NF-κB p65蛋白的表达无明显影响。结论:小檗碱可以改善高糖诱导的胰岛素抵抗,其分子机制可能与小檗碱抑制NF-κB p65的核转位有关。
Objective: To investigate the effect of berberine on the expression and translocation of NF-κB p65 in insulin-resistant model of 3T3-L1 adipocytes induced by high glucose, and to explore the molecular biological mechanism of berberine in improving insulin resistance. METHODS: Insulin resistance was induced in 3T3-L1 adipocytes by 25 mmol·L-1 glucose plus 0.6nmol·L-1 insulin, and berberine was used for intervention. Simultaneously, aspirin was used as a positive control, 2-deoxy-[3H]-D. Glucose uptake method was used to calculate the glucose transport rate. The expression of NF-κB p65 protein and nuclear NF-κB p65 protein in 3T3-L1 adipocytes was detected by Western blot. NF-κB p65 protein was localized by laser scanning confocal microscopy (CLSM). display. RESULTS: After 25 mmol·L-1 glucose plus 0.6 nmol·L-1 insulin for 18 h, insulin-stimulated glucose transport rate in 3T3-L1 adipocytes was inhibited by 60%. Western blot showed a significant increase in nuclear NF-κB p65 protein expression. CLSM showed that Nuclear translocation of NF-κB p65 increased; simultaneous addition of berberine or aspirin reversed these effects. However, high glucose, berberine, and aspirin had no significant effect on the expression of total NF-κB p65 protein in 3T3-L1 adipocytes. CONCLUSION: Berberine can improve insulin resistance induced by high glucose, and its molecular mechanism may be related to the inhibition of nuclear translocation of NF-κB p65 by berberine.