邹佳华,余昭胜,邓光锐,等.X射线照射后活性氧对巨噬细胞分泌肿瘤坏死因子-α和转化生长因子-β1的影响[J].安徽医药,2018,22(8):1444-1448. |
X射线照射后活性氧对巨噬细胞分泌肿瘤坏死因子-α和转化生长因子-β1的影响 |
ROS-induced expression of TNF-α and TGF-β1 in the RAW264.7 macrophage cell after X-ray radiation |
投稿时间:2017-05-27 |
DOI: |
中文关键词: 巨噬细胞 X射线 活性氧 细胞因子 |
英文关键词: Macrophage cell X-ray Reactive oxygen species Cytokine |
基金项目:湖北省自然科学基金(2014CFB423) |
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中文摘要: |
目的 研究X射线照射后活性氧(reactive oxygen species,ROS)对巨噬细胞肿瘤坏死因子-α(TNF-α)和转化生长因子-β1(TGF-β1)的影响。 方法 采用X射线照射巨噬细胞,收集细胞培养上清液,酶联免疫吸附实验(ELISA)法检测RAW264.7细胞TNF-α和TGF-β1。采用X射线照射巨噬细胞,荧光探针法检测ROS。随后采用活性氧清除剂N-乙酰半胱氨酸(NAC)和TNF-α抑制剂Lenalidomide预处理巨噬细胞后照射X射线,检测ROS, TNF-α和TGF-β1。采用Western blot检测Nox2的表达。结果 X射线照射后细胞培养上清液中ROS在照射后12 h达到最高,TNF-α在照射后24 h达到最高,TGF-β1在照射后48 h达到最高。使用3 mmol·L-1 NAC后,ROS明显受到抑制,同时TNF-α, TGF-β1分泌减少。使用2 μmol·L-1 Lenalidomide后,TNF-α分泌明显减少,同时TGF-β1分泌减少,ROS未受到影响。X射线照射巨噬细胞后Nox2的表达显著上升。结论 X射线照射巨噬细胞激活Nox2导致ROS增加,进而激活炎性因子TNF-α表达,最终导致TGF-β1表达升高。 |
英文摘要: |
Objective To investigate the regularity of reactive oxygen species induced secretion(ROS) of TNF-α and TGFβ1 in the RAW264.7 macrophage cell after X-ray radiation. Methods RAW264.7 macrophage cells were irradiated by X-ray. Then cell supernatant was collected. The secretion of TNF-α and TGFβ1 were assayed by ELISA. The level of ROS were assayed by nuorescent probe. The level of TNF-α, TGFβ1 and ROS were assayed after RAW264.7 macrophage cell pre-treated by NAC and Lenalidomide irradiated by X-ray. The expression of Nox2 were assayed by Western blot. Results The level of ROS reached a peak secretion after irradiation on 12 h. The level of TNF-α reached a peak secretion after irradiation on 24 h. The level of TGFβ1 reached a peak secretion after irradiation on 48 h. The level of ROS was inhibited in the RAW264.7 cell pre-treated by 3mM NAC after X-ray radiation.At the same time the level of TNF-α and TGF-β1 declined. The level of TNF-α was inhibited in the RAW264.7 cell pre-treated by 2 μmol·L-1 Lenalidomide after X-ray radiation,meanwhile, the level of TGF-β1 was declined and the level of ROS was not affected. The expression of Nox2 was significantly increased after X-ray radiation. Conclusion After X-ray radiation, macrophage cells activated Nox2 and led to increase of ROS, which further activated the expression of inflammatory cytokines TNF-α and lead to increased expression of TGF-β1. |
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