脱氧雪腐镰刀菌烯醇诱导CTLL-2细胞凋亡的信号途径研究

发布时间：2018-11-22 16:11

【摘要】：为探讨脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)对动物免疫功能的影响及其机理,以细胞毒性T淋巴细胞株CTLL-2为材料,用不同浓度的DON(0、0.5、1、2μg/m L)处理CTLL-2细胞48h,流式细胞术检测细胞的凋亡率,免疫印记法检测细胞凋亡相关通路蛋白CleavedCaspase 9、Cleaved-Caspase 3、Fas L、Fas、Cleaved-Caspase8等表达情况;并用流式细胞术检测了DON(1μg/m L)与Fas L中和抗体(10μg/m L)共处理后细胞的凋亡率。结果表明,随着DON浓度的升高,CTLL-2细胞的凋亡率呈上升趋势,染毒组均较对照组有极显著差异(P0.01);Bax/Bcl-2的比值升高,呈剂量-效应关系;DON促进细胞色素c(Cyt-c)从线粒体中释放到胞浆中,能够上调Cleaved-Caspase 9、Cleaved-Caspase 3的表达,染毒组均较对照组有显著性或极显著差异(P0.05或P0.01);Fas L、Fas、Cleaved-Caspase 8蛋白表达量随着DON浓度的升高逐渐增加,染毒组均较对照组有显著性或极显著差异(P0.05或P0.01);随着DON浓度的升高,p38 MAPK、JNK、Erk的磷酸化水平也升高,呈剂量-效应关系;与DON处理组相比,Fas L中和抗体与DON共同处理组细胞凋亡率明显下降(P0.01)。结果表明,DON可以诱导CTLL-2细胞发生凋亡,激活CTLL-2细胞线粒体通路、MAPKs通路,DON通过死亡受体途径诱导CTLL-2细胞凋亡,影响动物的免疫机能。
[Abstract]:In order to investigate the effect of deoxynivalenol (deoxynivalenol,DON) on animal immune function and its mechanism, CTLL-2 cells were treated with different concentrations of DON (0. 5% 1 渭 g / mL) for 48 h with cytotoxic T lymphocyte line CTLL-2. The apoptosis rate was detected by flow cytometry, and the expression of Cleaved-Caspase8 was detected by immunoblotting, and the expression of Cleaved-Caspase8 was detected by CleavedCaspase _ 9 Cleaved-Caspase3, and the expression of Cleaved-Caspase8 was detected by flow cytometry. The apoptosis rate of DON (1 渭 g / mL) co-treated with Fas L neutralizing antibody (10 渭 g / mL) was determined by flow cytometry. The results showed that the apoptotic rate of CTLL-2 cells increased with the increase of DON concentration, and the ratio of Bax/Bcl-2 increased in a dose-effect relationship between the exposed group and the control group (P0.01). DON promoted the release of cytochrome c (Cyt-c) from mitochondria to cytoplasm and up-regulated the expression of Cleaved-Caspase 9 Cleaved-Caspase 3, which was significantly or extremely different from that of control group (P0.05 or P0.01). The expression of Cleaved-Caspase 8 protein increased with the increase of DON concentration, and the expression of Cleaved-Caspase 8 protein in the exposed group was significantly higher than that in the control group (P0.05 or P0.01). With the increase of DON concentration, the phosphorylation level of p38 MAPK,JNK,Erk increased in a dose-effect relationship, and the apoptosis rate of, Fas L neutralizing antibody and DON co-treated with DON decreased significantly (P0.01). The results showed that DON could induce apoptosis of CTLL-2 cells, activate mitochondrial pathway and MAPKs pathway of CTLL-2 cells, and DON could induce apoptosis of CTLL-2 cells through death receptor pathway, which affected the immune function of animals.
【作者单位】：扬州大学兽医学院;江苏省动物重要疫病与人兽共患病防控协同创新中心;
【基金】：江苏省大学生实践创新训练计划项目(201511117043Z) 江苏高校优势学科建设工程资助项目(PAPD) 江苏省普通高校自然科学基金(08KJD230002)
【分类号】：S859.8

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