蒲黄总黄酮抑制棕榈酸培养下C2C12骨骼肌细胞白细胞介素6的表达

Pollen Typhae total flavones inhibit expression of interleukin-6 in C2C12 skeletal muscle cells cultured with palmitate

目的:观察蒲黄总黄酮(Pollen Typhae total flavones,PTF)对棕榈酸培养下C2C12骨骼肌细胞白细胞介素6(interleukin-6,IL-6)表达的影响,探讨PTF改善骨骼肌胰岛素抵抗(insulin resistance,IR)的可能机制。方法:0.25mmol/L棕榈酸培养建立骨骼肌细胞IR模型。根据处理方法不同,设对照组、模型组、核转录因子κB(nuclear factor-kappaB,NF-κB)抑制剂PDTC组、罗格列酮(rosiglitazone,ROS)组、ROS+PDTC组、PTF组和PTF+PDTC抑制剂组。处理16h后,3H-脱氧葡萄糖摄入法观察细胞对葡萄糖的转运率,实时定量多聚酶联反应检测IL-6mRNA的表达,酶联免疫吸附测定法检测细胞培养上清中IL-6蛋白水平。结果:0.25mmol/L棕榈酸培养16h,C2C12细胞葡萄糖摄取率下降30.43%,IR模型建立;PTF可使IR模型细胞葡萄糖转运率增加32.39%,IL-6mRNA表达及细胞培养上清液中IL-6蛋白水平均显著下降,差异有统计学意义(P<0.05);加入PDTC后,细胞IL-6mRNA表达及细胞培养上清液中IL-6蛋白水平上升(P<0.05)。结论:PTF通过抑制NF-κB通路而抑制C2C12骨骼肌细胞IL-6mRNA表达及蛋白分泌,可能是其减轻骨骼肌炎症状态和改善IR的机制之一。

Objective： To observe the effects of Pollen Typhae total flavones （PTF） on expression of interleukin-6 （IL-6） mRNA and protein secretion in C2C12 cell strain of skeletal muscle cells cultured with palmitate, and to explore the mechanism of PTF in relieving insulin resistance （IR）. Methods： The IR of C2C12 cells was induced by co-culturing with palmitate. The C2C12 cells were divided into normal control group, untreated group, PDTC （a nuclear factor-kappaB inhibitor） treated group, rosiglitazone （ROS）-treated group, ROS＋ PDTC -treated group, PTF-treated group and PTF＋PDTC-treated group. Sixteen hours after culture, the transportation rate of glucose was observed by ^3H-deoxygiucose uptake method; IL-6 mRNA expression in C2C12 cells was assayed by real time potymerase chain reaction （Real-time PCR）, and level of IL-6 protein secretion in culture supernatant was detected by enzyme linked immunosorbent assay. Results： Compared with the normal control group, the transportation rate of glucose of cells in untreated group was decreased 30. 43% after 16-hour palmitate culture, and was increased 32. 39% in the PTF-treated group. Compared with the untreated group, the levels of IL-6 mRNA expression in cells and IL-6 protein secretion in supernatant were significantly decreased in the PTF-treated group （P〈0. 05）. The levels of IL-6 mRNA expression in cells and IL-6 protein secretion in supernatant were increased in PTF＋ PDTC-treated group as compared with PFT-treated group （P〈0. 05）. Conclusion： PTF can inhibit the IL-6 mRNA expression and IL-6 protein secretion via nuclear factor-kappaB pathway in C2C12 skeletal muscle cells, which may be one of its mechanisms in relieving inflammation conditions and insulin resistance in C2C12 skeletal muscle cells.