氧化石墨烯衍生物对L6细胞活性的影响

摘要

目的：研究氧化石墨烯(GO)衍生物对L6细胞活性的影响。方法原代的L6细胞经过1 d或3 d培养，倒置荧光显微镜观察细胞形态并采取电子图片保存；收集对数期成熟的L6细胞，MTT法测定细胞增殖活性；对数期成熟的L6细胞，经GO衍生物3 d的刺激培养，收集细胞培养上清液，采用ELISA方法测定肿瘤坏死因子-α(TNF-α)和干扰素-γ(IFN-γ)。结果原代L6经过3 d培养，80%以上细胞分化为成熟的L6细胞；0.3μg/mL GO衍生物(GO-L和GO-D)经过3 d刺激，GO-L刺激细胞增殖的OD值为(0.52±0.28)，GO-D刺激细胞增殖的OD值为(0.81±0.34)，两者与对照细胞OD值(0.18±0.06)比较差异有显著统计学意义(F=6.05，P<0.01)；其中D型衍生物对L6细胞增殖活性强于L型衍生物，两者之间比较差异有统计学意义(F=4.54，P<0.05)；L型衍生物和D型衍生物均能诱导L6细胞分泌TNF-α[(18.6±5.36) vs (24.9±8.86)]和少量IFN-γ[(14.3±3.78) vs (13.8±2.62)]。但IFN-γ分泌两者差异无统计学意义(F=0.82，P>0.05)。结论 GO衍生物能诱导L6细胞活化和分泌TNF-α，可能会影响L6细胞中糖的代谢与调节。%Objective To investigate the effect of graphene oxide (GO) derivatives on the activity of L6 cells. Methods Morphologies of primary L6 cells, which had been cultured for 1 day or 3 days, and the cell morphology were observed with inverted fluorescent microscope and saved as electronic graphics. Mature L6 cells in logarithmic phase were collected. Cell proliferation activity was detected by MTT assay. Mature L6 cells in logarithmic phase stimu-lated with GO derivatives had been cultured for 3 days, then cell culture supernatants were collected. The serum tumor necrosis factor alpha (TNF-α) and interferon gamma (INF-γ) levels were determined by ELISA. Results Over 80%of primary L6 cells had been cultured for 3 days differentiated into mature ones. 0.3 μg/mL GO derivatives (GO-L and GO-D) were stimulated by 3 d. OD value for cell proliferation stimulated by GO-L and GO-D was (0.52 ± 0.28), (0.81 ± 0.34), respectively. Both had significant differences compared with OD value for control cells [(0.18 ± 0.06), F=6.05, P<0.01]. Activity of GO-D on L6 cells proliferation was significantly higher than GO-L (F=4.54, P<0.05). Both GO-L and GO-D could induce L6 cells to secrete TNF-α[(18.6±5.36) and (24.9±8.86) respectively] and a small amount of IFN-γ [(13.8 ± 2.62) and (14.3 ± 3.78) respectively]. But there were no significant differences in IFN-γ between the GO-L and GO-D (F=0.82, P>0.05). Conclusion GO derivatives can induce L6 cell activation and secretion of TNF-α, both of which may affect the metabolism and regulation of sugar in L6 cells.