交通相关PM2.5通过钙信号通路调控Jurkat T细胞IL-2的释放

发布时间：2018-12-14 03:04

【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(P0.05),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(P0.05)。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(P0.05)。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。
[Abstract]:Objective to investigate the effect of traffic related PM2.5 on interleukin-2 (IL-2) in Jurkat T cells and the regulation of calcium signaling pathway on IL-2 release. Methods Jurkat T cells were exposed to 100 渭 g/ml PM2.5, and normal saline was used as control group. The calcium chelator (EGTA), a calcium chelator of cyclosporin A (CSA), was used as a calcium chelating agent. The blank filter membrane group was set up. The levels of IL-2 in supernatant of EGTA group and CSA group were measured by 3 h.ELISA and 24 h.ELISA methods. QRT-PCR was used to detect the mRNA expression of T cell transcription factor (NFAT) activated by calmodulin (CaN), and the distribution of NFAT nucleus was observed by immunofluorescence. Results the level of IL-2 in 100 渭 g/ml PM2.5 group was significantly lower than that in blank filter membrane and saline group, but higher than that in 100 渭 g/ml PM2.5 CSA group and 100 渭 g/ml PM2.5 EGTA group. The difference was statistically significant (P0.05), and the expression level of IL-2 decreased with time. The NFAT,CaN mRNA expression level of 100 渭 g/ml PM2.5 group was higher than that of control group, 100 渭 g/ml PM2.5 CSA group. In the 100 渭 g/ml PM2.5 EGTA group, the difference was statistically significant (P0.05). PM2.5 could dephosphorize and activate the NFAT protein, which could be translocated into the nucleus. The expression level of IL-2 was negatively correlated with NFAT gene and CaN gene expression, and the difference was statistically significant (P0.05). Conclusion PM2.5 can inhibit the release of IL-2,Ca2-CaN-NFAT from Jurkat T cells and may be involved in the regulation of IL-2 release from Jurkat T cells by PM2.5.
【作者单位】：山西医科大学公共卫生学院环境卫生学教研室;
【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
【分类号】：R392

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