Lou Shao-Ying, Liu Yi, Chen Wei-Hua, Ying Jian, He Yan-Ming, Wang Wen-Jian
Institution of Chinese Integrative Medicine, Huashan Hospital, Fudan University, Shanghai 200040, China.
Zhong Xi Yi Jie He Xue Bao. 2008 May;6(5):488-92. doi: 10.3736/jcim20080511.
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).
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.
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).
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.
观察蒲黄总黄酮(PTF)对棕榈酸培养的骨骼肌C2C12细胞株白细胞介素-6(IL-6)mRNA表达及蛋白分泌的影响,探讨PTF改善胰岛素抵抗(IR)的机制。
通过与棕榈酸共培养诱导C2C12细胞产生IR。将C2C12细胞分为正常对照组、未处理组、PDTC(核因子-κB抑制剂)处理组、罗格列酮(ROS)处理组、ROS+PDTC处理组、PTF处理组和PTF+PDTC处理组。培养16小时后,采用(3)H-脱氧葡萄糖摄取法观察葡萄糖转运率;采用实时聚合酶链反应(Real-time PCR)检测C2C12细胞中IL-6 mRNA表达,采用酶联免疫吸附测定法检测培养上清液中IL-6蛋白分泌水平。
与正常对照组相比,未处理组细胞经16小时棕榈酸培养后葡萄糖转运率降低30.43%,PTF处理组升高32.39%。与未处理组相比,PTF处理组细胞中IL-6 mRNA表达水平及上清液中IL-6蛋白分泌水平均显著降低(P<0.05)。与PTF处理组相比,PTF+PDTC处理组细胞中IL-6 mRNA表达水平及上清液中IL-6蛋白分泌水平升高(P<0.05)。
PTF可通过核因子-κB途径抑制C2C12骨骼肌细胞中IL-6 mRNA表达及IL-6蛋白分泌,这可能是其改善C2C12骨骼肌细胞炎症状态及胰岛素抵抗的机制之一。