摘要
目的探讨交通相关PM2.5对Jurkat T细胞白介素-2(IL-2)的影响及钙信号通路对IL-2释放的调控作用。方法以100μg/ml的PM2.5染毒Jurkat T细胞,以生理盐水为对照组,并用钙调磷酸酶拮抗剂环孢菌素A(CSA)、钙离子螯合剂(EGTA)进行干预,设置空白滤膜组、EGTA组和CSA组平行对照,染毒时间为3、6和24 h。ELISA方法测定细胞上清IL-2水平;QRT-PCR测定细胞钙调磷酸酶(CaN)、活化T细胞转录因子(NFAT)的mRNA表达;免疫荧光法观察NFAT核分布情况。结果交通相关PM2.5染毒细胞3、6和24 h后,100μg/ml PM2.5组IL-2水平明显低于空白滤膜和生理盐水组,但高于100μg/ml PM2.5+CSA组、100μg/ml PM2.5+EGTA组,差异均有统计学意义(P<0.05),并且随着时间的延长,IL-2表达水平呈降低趋势。100μg/ml PM2.5组的NFAT、CaN mRNA表达水平高于对照组、100μg/ml PM2.5+CSA组、100μg/ml PM2.5+EGTA组,差异均具有统计学意义(P<0.05)。PM2.5能够使NFAT蛋白脱磷酸化而活化,可移位至细胞核内。白介素-2表达水平与NFAT基因、CaN基因表达水平呈负相关,差异均具有统计学意义(P<0.05)。结论交通相关PM2.5可抑制Jurkat T细胞释放IL-2,Ca2+-CaN-NFAT信号通路可能参与调控PM2.5对Jurkat T细胞IL-2的释放。
Objective To explore the effects of traffic-related PM2.5 on interleukin-2 （IL-2） in Jurkat T cells and the regulatory action of calcium signaling pathway. Methods The cells were exposed to 100 μg/ml of PM2.5 for 3, 6 and 24 h. Normal saline group, blank filter group, calcium chelating agent EGTA group and the calcineurin antagonist eyclosporine A （CSA） group were as parallel control. The level of IL-2 was detected by ELISA kits, the mRNA expression of CaN, NFAT were determined by QRT- PCR. The nuclear distribution of NFAT was observed by immunofluorescence microscopy. Results The level of IL-2 in Jurkat T cells exposed to 100 μg/ml PM2. 5 was significantly lower than parallel groups, but higher than PM2. 5 ＋ CSA group and PM2. 5 ＋ EGTA group （P 〈 0. 05）. With the increase of time, the releasing level of IL-2 appeared reducing trend in 100 μg/ml of PM2.5 group. The mRNA expression level of NFAT and CaN were higher than parallel groups, PM2.5 ＋ CSA group and PM2.5 ＋ EGTA group （P 〈 0.05）. PM2.5 can induce NFAT protein with dephosphorylation and be activated, and NFAT protein can shift into nuclear. The level of IL-2 was negatively associated with the expression level of NFAT and CaN gene（P 〈 0.05）. Conclusion Traffic-related PM2.5 may inhibit the releasing of IL-2, Ca^2＋-CaN-NFAT signal pathway may involve in the regulation of IL-2.
出处
《卫生研究》
CAS
CSCD
北大核心
2013年第5期822-827,共6页
Journal of Hygiene Research
基金
国家自然科学基金(No.81072261)
2009年山西省高校优秀青年学术带头人支持项目
