Zhou Hui-Ren, Islam Zahidul, Pestka James J
Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing Michigan 48824-1224, USA.
Toxicol Sci. 2005 Sep;87(1):113-22. doi: 10.1093/toxsci/kfi234. Epub 2005 Jun 23.
Deoxynivalenol (DON) and other ribotoxic trichothecenes cause immune stimulation and suppression in leukocytes by upregulating gene expression and apoptosis, respectively. The purpose of this study was to test the hypothesis that MAPKs mediate both apoptosis and survival in DON-exposed macrophages. At concentrations which partially inhibit translation, DON induced phosphorylation of p38 and ERK 1/2 mitogen activated protein kinases within 15 min in RAW 264.7 macrophages and these effects lasted up to 3 h. DON-exposed cells exhibited marked caspase 3-dependent DNA fragmentation after 6 h which was suppressed and attenuated by the p38 inhibitor SB203580 and ERK inhibitor PD98059, respectively. DON readily induced the phosphorylation and activity of p53 and this was inhibitable by SB203580. DON exposure evoked BAX translocation to mitochondria and corresponding cytochrome C release but did not alter mitochondrial membrane potential. The p53 inhibitor PFTalpha reduced both DON-induced phosphorylation of p53 and p53 binding activity. Moreover, both PFTalpha and p53 siRNA transfection suppressed DON-induced caspase-3 activity and subsequent DNA fragmentation. Concurrent with p53 activation, DON activated two anti-apoptotic survival pathways as evidenced by both ERK-dependent p90 Rsk and AKT activation. Taken together, the results indicate that DON initiates competing apoptotic (p38/p53/Bax/Mitochondria/Caspase-3) and survival (ERK/AKT/p90Rsk/Bad) pathways in the macrophage.
脱氧雪腐镰刀菌烯醇(DON)和其他核糖体毒性单端孢霉烯族毒素分别通过上调基因表达和诱导细胞凋亡,从而引起白细胞的免疫刺激和抑制。本研究的目的是验证丝裂原活化蛋白激酶(MAPKs)介导DON暴露巨噬细胞凋亡和存活的假说。在部分抑制翻译的浓度下,DON在15分钟内诱导RAW 264.7巨噬细胞中的p38和ERK 1/2丝裂原活化蛋白激酶磷酸化,且这些效应持续长达3小时。暴露于DON的细胞在6小时后表现出明显的半胱天冬酶3依赖性DNA片段化,分别被p38抑制剂SB203580和ERK抑制剂PD98059抑制和减弱。DON容易诱导p53的磷酸化和活性,且这可被SB203580抑制。DON暴露引起BAX转位至线粒体及相应的细胞色素C释放,但未改变线粒体膜电位。p53抑制剂PFTα降低了DON诱导的p53磷酸化和p53结合活性。此外,PFTα和p53 siRNA转染均抑制了DON诱导的半胱天冬酶3活性及随后的DNA片段化。与p53激活同时发生的是,DON激活了两条抗凋亡存活途径,ERK依赖性p90Rsk和AKT激活证明了这一点。综上所述,结果表明DON在巨噬细胞中启动了相互竞争的凋亡(p38/p53/Bax/线粒体/半胱天冬酶3)和存活(ERK/AKT/p90Rsk/Bad)途径。