摘要
目的探讨X射线对小鼠小胶质BV-2细胞极化的影响。方法取对数生长期BV-2细胞随机分为假照射组(Sham)和10 Gy照射组,后者给予X射线单次照射,剂量为1.28 Gy/min,照射时间7 min 49 s。于照射后1、3、6、24 h和48 h,光镜下观察并分析BV-2细胞活化率;HE染色及Iba-1免疫荧光染色观察BV-2细胞形态;ELISA检测BV-2细胞培养液上清中M1型活化标志物TNF-α、IL-1β和M2型活化标志物TGF-β1水平变化;Western blotting检测BV-2细胞M1型(CD86、iNOS)和M2型(CD206)极化相关蛋白水平变化。结果形态学结果显示,10 Gy X射线照射后细胞出现胞体变大、突起变粗变短,呈"阿米巴"样改变。与Sham组相比,10 Gy照射后3 h BV-2细胞活化率即明显升高,并在照后6 h达到高峰,48 h开始恢复;ELISA结果显示,TNF-α和TGF-β1水平在照后48 h时均明显升高;IL-1β水平在照后3~6 h出现一过性降低趋势,在照后24 h开始增高,至照后48 h达到高峰;Western blotting结果显示,CD86蛋白水平在照后各时间点无显著变化,iNOS蛋白表达水平在照后1、6、24 h和48 h显著增高;CD206在照后3 h出现先降低后升高交替波动性变化。结论10 Gy X射线单次照射可诱导离体培养的BV-2细胞激活,其极化类型随照后时间的不同而变化。
Objective To investigate the effect of X-ray on the polarization of mouse microglia BV-2 cells.Methods BV-2 cells at the logarithmic growth stage were randomly divided into the Sham irradiation group and 10 Gy irradiation group.The latter group was given a single X-ray irradiation at a dose of 1.28 Gy/min for 7 min 49 s.The activation rate of BV-2 cells was observed and analyzed under a microscope at 1,3,6,24 h and 48 h after irradiation.The changes of cell morphology were observed by HE staining and immunofluorescence staining;The levels of M1-type activation markers(TNF-αand IL-1β)and M2-type activation marker TGF-β1 in the supernatant of BV-2 cells were detected by ELISA.The levels of polarization-related proteins of M1-type(CD86 and iNOS)and M2-type(CD206)in BV-2 cells were detected by Western blotting.Results Morphological results showed that BV-2 cells became larger,and their protrude became coarse and shorter,showing"amoeba"like changes after 10 Gy X-ray irradiation.Compared with the Sham group,the activation rate of BV-2 cells was significantly increased at 3 h,and reached the peak at 6 h,and began to recover at 48 h after irradiation.ELISA results showed an obvious increase in the level of TNF-αand TGF-β148 h after irradiation.The level of IL-1βshowed a transient decrease at 3~6 h,increased at 24 h,and reached the peak 48 h after irradiation.Western blotting results showed that CD86 protein level did not change significantly at each time points after irradiation,and iNOS protein level increased significantly at 1,6,24 h and 48 h after irradiation.A fluctuating change in CD206 protein level was found after irradiation.Conclusion 10 Gy X-ray irradiation can induce the activation of BV-2 cells in vitro,and the polarization type changes with the time after irradiation.
作者
黄蓉蓉
周艳
郭玲
刘力源
薛一哲
来盼盼
荆云涛
徐惠
张钦凤
丁桂荣
HUANG Rongrong;ZHOU Yan;GUO Ling;LIU Liyuan;XUE Yizhe;LAI Panpan;JING Yuntao;XU Hui;ZHANG Qinfeng;DING Guirong(School of Public Health,Shandong First Medical University,Tai’an 271000 China;Department of Radiation Protection Medicine,Department of Military Preventive Medicine,Air Force Medical University,Xi’an 710032 China;College of Radiology,Shandong First Medical University,Tai’an 271000 China)
出处
《中国辐射卫生》
2021年第3期247-252,共6页
Chinese Journal of Radiological Health
基金
山东省自然科学基金(ZR2020MH328)。
关键词
电离辐射
X射线
小胶质细胞
M1极化
M2极化
Ionizing Radiation
X-ray
Microglia Cells
M1 Polarization
M2 Polarization
