氟对小胶质细胞炎性因子表达及细胞核因子κB信号通路的影响  

Enhanced expression of inflammatory cytokines and nuclear factor-KB in microglia by overdose fluoride

作　　者：汤婷婷[1] 禹文峰 官志忠[1] 

机构地区：[1]贵州医科大学病理学教研室,550004 [2]贵州省医学分子生物学重点实验室

出　　处：《中华地方病学杂志》2015年第11期785-789,共5页Chinese Journal of Endemiology

基　　金：国家自然科学基金（81160335）;科技部支撑计划课题（2011BAZ03220）;贵州省创新计划项目（[20141]6）;贵州省卫生厅项目（G2010-20）

摘　　要：目的观察氟对小胶质细胞炎症因子表达及细胞核因子KB（NF-κB）信号通路的影响。方法以体外培养人急性单核细胞白血病细胞（THP-1）作为小胶质细胞模型，用不同浓度[0（阴性对照组）、10、50、100、500、1000、5000μm0L／L]氟化钠（NaF）培养48h，CCK8法检测细胞存活率。根据CCK8法检测结果，将THP-1细胞分为3组：对照组和低、高剂量染氟组，分别用0、500、5000μmoVLNaF染氟处理48h．以酶联免疫吸附实验（ELISA）检测细胞培养液上清中炎性细胞因子白细胞介素（IL）-1β和肿瘤坏死因子（TNF）-α，蛋白质免疫印迹（Westemblot）法检测培养细胞IκBα、Phospho-NF-κBp65（P-P65）和Phospho-IKB．-α（P-IκBα）蛋白表达水平。结果500、1000、5000μmol／L染氟组THel细胞存活率[（73．21±3．67）％、（31．40±4．56）％、（0．40±0．24）％]明显低于阴性对照组[（100．00±0．00）％，P均〈0．01]。低、高剂量染氟组炎性因子IL-1β[（1．42±0．79）、（19．47±2．90）ng／L]、TNF-α含量[（61．06±2．20）、（172．72±2．29）ng／L]均高于对照组[（0．36±0．07）、（31．07±0．81）．g／L，P均〈0．05]；高剂量染氟组IκBα蛋白表达水平[（63．53±9．67）％]低于对照组[（100．00±10．99）％，P〈0．01]；低、高剂量染氟组P-P65蛋白表达水平[（113．71±8．99）％、（134．74±1．93）％]明显高于对照组[（100．00±5．48）％，P均〈0．05]；低、高氟剂量组P-IκBα蛋白表达水平[（152．61±14.16）％、（176．91±7．95）％]高于对照组[（100．00±14．82）％，P均〈0．01]。线性回归分析表明，TNF-α和IL-1β含量与P-P65蛋白表达水平之间呈线性正相关（r=0．74、0．72，P均〈0．05）。结论过量氟可诱导小胶质细胞产生炎性因子并激活NF．κB信号通路，炎性因子释放及信号通路的激活可能是氟导致中枢神经系统损伤的机制之一�Objective To investigate fluoride-induced inflammation and nuclear factor-KB （NF-KB） signaling pathway in cultured human acute monocytic leukemia cells （THP-1）. Methods In vitro cultured THP-1 ceils were used as a model of microglia. After cultured with different of [0 （negative control group）, 10, 50, 100, 500, 1 000 and 5 000 μmol/L] sodium fluoride （NaF） for 48 h, the survival of cells was detected by CCK8. THP-1 cells were divided into 3 groups： control group, low dose and high dose fluoride groups according to the results of CCK8 assay, and then treated with different concentrations of sodium fluoride （0, 500, 5 000 μmol/L） for 48 h, concentrations of inflammatory cytokines, such as Interleukin-ll3 （IL-1[β） and tumor necrosis factor-or （TNF--α） were measured by enzyme linked immunosorbent assay （ELISA） in THP-1 mononuclear cell culture medium. The protein levels of IKBα, phospho-NF-κB p65 and phospho-IκB-α were detected by Western blotting. Results THP-1 cells were treated with different concentrations of sodium fluoride （500, 1 000, 5 000 μmol/L） for 48 h. Fluoride group THP-1 cell survival rate [（73.21 ±3.67）%, （31.40 ± 4.56）%, （0.40 ± 0.24）%] was lower than that of the negative control group [（100.00 ± 0.00）%, all P 〈 0.01]. Compared to the control groups [（0.36 ± 0.07）, （31.07 ±0.81）ng/L[, significant increases of the inflammatory cytokines IL-1β [（1.42 ± 0.79）, （19.47 ±2.90）ng/L] and TNF-α [（61.06 ±2.20）, （172.72± 2.29）ng/L] were detected in culture medium in low-fluoride and high fluoride groups, respectively. Interestingly, compared to the control groups [（100.00 ±5.48）%, （100.00 ± 14.82）%], significant increases of phospho-NF-κB p65 [（113.71 ± 8.99）%, （134.74± 1.93）%] and phospho-IκB-α [（152.61 ±14.16）%, （176.91 ± 7.95）%] were observed in both low-fluoride and high fluoride groups. Meanwhile, the protein level of IκBoα in high fluoride group [（63.53 ± 9.67）% ] w

关 键 词：氟化物中毒 小胶质细胞 细胞核因子ΚB 

分 类 号：R599.1[医药卫生—内科学]