MAPK信号通路在交通相关PM2.5诱导CEM-6T细胞凋亡中作用

Effect of MAPK signaling pathway on CEM-6T cell apoptosis induced by traffic-related PM2.5

目的探讨MAPK信号通路在交通相关PM2.5诱导CEM-6T细胞凋亡中的作用,为研究交通相关细颗粒物的免疫毒性机制提供实验依据。方法采用0、20、80、320mg/L PM2.5染毒CEM-6T细胞24、48h,用Western blot法检测交通相关PM2.5对CEM-6T细胞p-ERK、p-JNK和p38蛋白的作用;用p-JNK的拮抗剂SP600125和p38的拮抗剂SB203580进一步反面验证交通相关PM2.5是否影响CEM-6T细胞p-JNK和p38蛋白的表达。采用单因素方差分析进行数据分析。结果用不同剂量PM2.5(0、20、80、320mg/L)染毒CEM-6T细胞24、48h,随着染毒剂量增加,p-ERK蛋白表达量逐渐减少,320mg/L PM2.5组p-ERK蛋白表达量和生理盐水组比较明显降低,差异有统计学意义(P<0.05)。p-JNK1和p-JNK2蛋白表达量随染毒剂量增加而增加,320mg/L组与生理盐水组比较差异有统计学意义(P<0.05)。p38蛋白表达量随染毒剂量增加而增加,p38蛋白表达量在80和320mg/L PM2.5染毒组与生理盐水组比较差异有统计学意义(P<0.05);染毒前加入JNK拮抗剂(SP600125)和p38拮抗剂(SB203580)后,pJNK2和p38蛋白表达量均较正常染毒时降低,加与未加拮抗剂320mg/L PM2.5染毒组比较,差异有统计学意义。结论交通相关细颗粒物可能通过MAPK信号通路诱导CEM-6T细胞凋亡。

ObjectiveTo explore the effect of MAPK signaling pathway on CEM-6T cell apoptosis induced by traffic related PM2.5, and to provide experimental evidence for understanding the mechanism of immunotoxic ity. Methods CEM 6T cells were exposed to PM2.5 at the doses of 0, 20, 80 and 320 mg/L for 24h or 48h. Western Blot was used to assess the effect of PM2.5 on p ERK, p-JNK and p38 inside cells. Meanwhile, the antagonists SP600125 for p JNK and SB203580 for p38 were also applied for further verification of the effect. One-way ANOVA was used for data statistical analysis. Results The p-ERK protein expression in CEM 6T cells decreased along with an increased exposure doses and time frames to traffic-related PM2.5. The lowest p-ERK protein level was noticed in cells exposed to the highest dose of 320 mg/L PM2.5 which had a significant difference compared to saline control （P〈0.05） . While, the p JNK1, p-JNK2 and p38 protein levels elevated in cells along with the increased doses of PM2.5 exposure. When cells were exposed to320 mg/L PM2.5, the differences of the above three protein levels were statistically significant compared to saline control （P-~0.0,5） . However, the addition of antagonists of p-JNK and p38 prior to the exposure could block the increases of p JNK2 and p38 protein levels, especially at the dose of 320mg/L. Conclusion Traffic related PM2.5 may induce CEM-6T cells apoptosis through the MAPK signaling pathway.