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-1NAC后,ROS明显受到抑制,同时TNF-α,TGF-β1分泌减少。使用2μmol·L-1Lenalidomide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。
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 3 m M 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.
作者
邹佳华
余昭胜
邓光锐
余胜年
王娴
韩秀文
汪井龙
付红梅
ZHOU Jiahua;YU Zhaosheng;DENG Guangrui;YU Shengnian;WANG Xian;HAN Xiuwen;WANG Jinglong;FU Hongmei(Tumor Center of Hubei Huanggang Traditional Chinese Medicine Hospital,Huanggang,Hubei 438000,China)
出处
《安徽医药》
CAS
2018年第8期1444-1448,共5页
Anhui Medical and Pharmaceutical Journal
基金
湖北省自然科学基金(2014CFB423)
关键词
巨噬细胞
X射线
活性氧
细胞因子
Macrophage cell
X-ray
Reactive oxygen species
Cytokine
