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表达.docVIP

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表达 　作者：娄少颖, 刘毅, 陈伟华, 应健, 何燕铭, 王文健 【摘要】 　　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：0.25 mmol/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16 h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：0.25 mmol/L棕榈酸培养16 h，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（Plt;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（Plt;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。 【关键词】 蒲黄总黄酮; 棕榈酸; 胰岛素抵抗; 白细胞介素6; C2C12细胞株 　　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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 s