脂联素通过AMPK调节HepG2细胞中支链氨基酸的代谢

杜朝升; 朱 迪; 邹 倩; 廉 坤; 洪志博; 周 芬; 陶 凌

脂联素通过AMPK调节HepG2细胞中支链氨基酸的代谢

Adiponectin improves branched-chain amino acid catabolism of HepG2 cells via upregulation of AMPK

摘要

摘要: 目的:探讨脂联素（APN）对HepG2细胞支链氨基酸（BCAA）分解代谢的影响及其作用机制。方法：将培养的HepG2细胞系随机分为对照组、APN组、APN+腺苷酸活化蛋白激酶(AMPK)抑制剂(Compound C，CpC)组(APN+CpC)组和AMPK激动剂（AICAR）组。用Western blot检测AMPK、P-AMPK、BCKD E1α（BCAA分解代谢过程中的关键酶，磷酸化时为失活状态）及P-BCKD E1α蛋白的表达，用ELISA方法检测培养HepG2细胞上清中BCAA的含量。结果：与对照组相比，APN组和AICAR组均可以显著上调细胞P-AMPK的水平（P<0.05，P<0.01），显著降低细胞p-BCKD E1α及上清中BCAA的水平（P<0.05，P<0.01）。与APN组比较，APN+CpC组，其细胞内P-AMPK水平显著下降（P<0.01），P-BCKD E1α及上清中BCAA的水平明显上升（分别为P<0.05及P<0.01）。结论： APN可显著促进HepG2细胞BCAA的分解代谢，其作用机制与AMPK磷酸化作用的上调相关。

Abstract: AIM:To investigate the promoter effect of adiponectin (APN) on the branched-chain amino acid (BCAA) catabolism of HepG2 cells and the underlying mechanism. METHODS: HepG2 cells randomly received a control, APN, APN+CpC and AICAR. Expressions of protein p-AMPK, AMPK, p-BCKD E1α and BCKD E1α were analyzed by Western blot and the BCAA level of supernatant from cultured HepG2 was detected by ELISA. RESULTS: Compared with that of vehicle, the supernatant BCAA level from cultured HepG2 significantly decreased (P<0.01) when APN or AICAR was administered, but the expression of p-AMPK protein from cells increased (P<0.01). Administration of CpC to HepG2 cells increased the supernatant BCAA level (P<0.01), but decreased the p-AMPK expression (P<0.01). CONCLUSION: Adiponectin notably improves the BCAA catabolism of HepG2 cells through upregulating p-AMPK expression.

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