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

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PTF4-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-6 mRNA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-6 mRNA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-6 mRNA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-6 mRNA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。

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

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 (3)H-deoxyglucose uptake method; IL-6 mRNA expression in C2C12 cells was assayed by real time polymerase 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.