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目的探讨肥胖启动炎症的分子机制。方法将3T3-L1前脂肪细胞诱导分化为成熟脂肪细胞,取诱导分化第8天的脂肪细胞,分为空白对照组、激动剂组和抑制剂组。空白对照组将高糖DMEM培养基作用于诱导分化的脂肪细胞;激动剂组将含0.5 mmol/L 5氨基-4-甲酰胺咪唑核糖核苷酸(AICAR)的高糖DMEM培养基作用于脂肪细胞1 h;抑制剂组将含10μmol/L Compound C的高糖DMEM培养基作用于脂肪细胞1 h,收集蛋白及培养液于-80℃保存。Western blot法检测磷酸化AMPK(p-AMPK)及p-NF-κB表达。酶联免疫吸附试验法测定IL-6与TNF-α水平。结果与空白对照组比较,激动剂组p-AMPK蛋白表达明显升高,p-NF-κΒ蛋白表达明显降低;而抑制剂组p-AMPK蛋白表达明显降低,p-NF-κΒ蛋白表达明显升高,P均<0.05。与空白对照组比较,激动剂组TNF-α与IL-6表达均明显降低,而抑制剂组TNF-α与IL-6表达均明显升高,P均<0.05。结论 AMPK可抑制NF-κΒ信号及炎症因子的分泌,肥胖导致炎症可能与肥胖时AMPK活性降低引发NF-κΒ信号活化增强有关。
Objective To investigate the molecular mechanism of obesity-induced inflammation. Methods 3T3-L1 preadipocytes were induced to differentiate into mature adipocytes. Adipocytes were induced to differentiate into adipocytes on day 8 and divided into blank control group, agonist group and inhibitor group. In the blank control group, high glucose DMEM medium was used to induce differentiation of adipocytes. In the agonist group, high glucose DMEM medium containing 0.5 mmol / L 5-amino-4-formamidazoleimidazole ribonucleotide (AICAR) Cells for 1 h. The inhibitor group was treated with high glucose DMEM containing 10 μmol / L Compound C for 1 h, and the protein and culture solution were collected and stored at -80 ℃. Western blot was used to detect the expression of phosphorylated AMPK (p-AMPK) and p-NF-κB. The levels of IL-6 and TNF-α were measured by enzyme linked immunosorbent assay. Results Compared with the blank control group, the expression of p-AMPK protein was significantly increased and the expression of p-NF-κB protein was significantly decreased in the agonist group; however, the expression of p-AMPK protein was significantly decreased and the expression of p-NF-κB protein was significantly increased High, P <0.05. Compared with the blank control group, the expression of TNF-α and IL-6 in the agonist group were significantly decreased, while the expression of TNF-α and IL-6 in the inhibitor group were significantly increased, P <0.05. Conclusion AMPK can inhibit the secretion of NF-κB signal and inflammatory cytokines. Obesity-induced inflammation may be related to the decrease of AMPK activity and the activation of NF-κB signaling during obesity.