葫芦巴4-羟基异亮氨酸对高糖诱导小鼠3T3-L1脂肪细胞胰岛素抵抗的影响  

Effect of Trigonella Foenum-graecum 4-Hydroxyisoleucine on High-glucose Induced Insulin Resistance in 3T3-L1 Adipocytes of Mice

作　　者：余海[1] 吴萌[2] 卢芙蓉[1] 谢晶[1] 郑娜[1] 秦铀[3] 高峰[4] 杜文[4] 剪柳萌[4] 

机构地区：[1]华中科技大学同济医学院附属协和医院中西医结合科,武汉430022 [2]华中科技大学同济医学院附属协和医院呼吸内科,武汉430022 [3]华中科技大学同济医学院附属协和医院肿瘤中心,武汉430022 [4]华中科技大学同济医学院附属协和医院内分泌科,武汉430022

出　　处：《中国中西医结合杂志》2013年第10期1394-1399,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81001670;81202962);华中科技大学同济医学院附属协和医院院内基金项目(No.2010-17)

摘　　要：目的研究葫芦巴活性成分4-羟基异亮氨酸(4-hydroxyisoleucine,4-HIL)对高糖诱导的小鼠前脂肪细胞(3T3-L1)胰岛素抵抗的作用及其机制。方法以25 mmol/L葡萄糖加0.6 nmol/L胰岛素诱导小鼠3T3-L1脂肪细胞建立胰岛素抵抗细胞模型,予以不同浓度4-HIL(5、10、20μmol/L)进行干预。采用[3H]-脱氧-D-葡萄糖摄入法检测葡萄糖摄取率,PCR法检测肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)mRNA表达量,ELISA法检测细胞上清TNF-α蛋白水平。以棕榈酸(palmitic acid,PA)为对照。结果 25mmol/L葡萄糖加0.6 nmol/L胰岛素作用18 h后,3T3-L1脂肪细胞胰岛素刺激的葡萄糖转运抑制了63%;PCR显示脂肪细胞内TNF-αmRNA的表达较正常脂肪细胞明显增高(P<0.05);ELISA显示细胞上清中的TNF-α分泌量增加70 pg/mL(P<0.05)。PA组表现出类似结果。分别加入5、10、20μmol/L 4-HIL作用24h后,3T3-L1脂肪细胞胰岛素刺激所产生的葡萄糖转运分别增加35%、50%、60%;PCR检测显示,脂肪细胞内TNF-αmRNA的表达随着药物浓度的增加呈递减趋势,与模型组及PA组比较,差异有统计学意义(P<0.05);与模型组比较,药物干预后脂肪细胞培养基上清TNF-α分泌水平分别下降10、18、39 pg/mL(P<0.05)。结论 4-HIL可以显著改善高糖诱导的小鼠3T3-L1脂肪细胞胰岛素抵抗,同时4-HIL能够抑制TNF-α的分泌。Objective To investigate the effect of 4-hydroxyisoleucine （4-HIL）, an active component of Trigonella Foenum-graecum L. on high glucose induced insulin resistance （IR） in 3T3-L1 adipo- cytes, and to explore underlying molecular mechanisms. Methods 3T3-L1 adipocytes were treated with 25 mmol/L glucose and 0.6 nmol/L insulin to induce IR. They were intervened by different concentrations of 4-HIL （at 5, 10, and 20 μmol/L）. E3Hl-Deoxy-D-glucose up-taking method was used to detect the glucose uptake. The mRNA expression of cellular tumor necrosis factor-alpha （TNF-cx） was detected by pol- ymerase chain reaction （PCR）. The content of TNF-e in the culture supernatant was detected by enzyme-linked immunosorbent assay （ELISA）. Palmitic acid （PA） acted as the control. Results After inter- vened by 25 mmol/L glucose and 0.6 nmol/L insulin for 18 h, the insulin-stimulated glucose transportation in 3T3-L1 adipocytes was inhibited by 63%. The mRNA expression of cellular TNF-oL in adipocytes signifi- cantly increased, when compared with that in normal adipocytes （P 〈0.05）. The level of TNF-oL secreted in the culture supernatant was increased by 70 pg/mL （P 〈0.05）. Similar changes occurred in the PA group. After exposure to 4-HIL （5, 10, or 20 IJ.mol/L） for 24 h, the glucose transportation was increased by 35%, 50%, and 60%, respectively. PCR results showed that along with increasing 4-HIL concentra- tions, the mRNA expression of cellular TNF-α showed a decreasing trend, showing statistical difference when compared with the model group and the PA group （P 〈0.05）. Compared with the model group, the TNF-α level in the supernatant was respectively reduced by 10 pg/mL, 18 pg/mL, and 39 pg/mL after intervention （P 〈0.05）. Conclusion 4-HIL could remarkably improve high glucose-induced IR in 3T3-L1 adipocytes. Meanwhile, 4-HIL could inhibit the secretion of TNF-α.

关 键 词：葫芦巴 4-羟基异亮氨酸 胰岛素抵抗 肿瘤坏死因子-Α 

分 类 号：R285.5[医药卫生—中药学]