| 摘要: |
| 【目的】探讨IBDV感染过程中凋亡相关基因启动子5-mC甲基化水平的变化，为揭示IBDV引起细胞凋亡的细胞内调节机制提供理论基础。【方法】采用21日龄母源性抗体检测为阴性的商品肉鸡，随机分为对照组（control）和病毒感染组（IBDV），病毒感染组肉鸡用IBDV CJ801毒液进行点眼滴鼻攻毒0.1 mL/只，对照组以同样方式给予PBS。IBDV感染后5 d收集法氏囊组织，采用HE染色观察法氏囊组织病理变化，采用qRT-PCR检测凋亡相关基因的表达，并应用MeDIP-PCR技术检测差异基因启动子的甲基化水平。【结果】与对照组相比，IBDV感染后5 d肉鸡法氏囊出现明显的组织水肿，滤泡面积明显降低，滤泡中出现大量固缩细胞核。促凋亡基因Caspase-8和Caspase-3的表达极显著降低（P＜0.01），抑制凋亡基因Bcl-2的表达极显著升高（P＜0.01）。MeDIP-PCR发现，Caspase-3基因启动子的甲基化水平显著升高（P＜0.05），而Bcl 2基因启动子的甲基化水平极显著降低（P＜0.01）。【结论】IBDV感染引起凋亡相关基因的表达可能与其启动子甲基化水平有关。 |
| 关键词: IBDV 细胞凋亡 凋亡基因 DNA甲基化 |
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| 基金项目:国家自然科学基金项目（31602024，31702199）；河南省教育厅重点科研计划项目（17A230004，17B230002） |
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| IBDV induced changes in expression of apoptosis-related genes |
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GUO Feng,WU Qi,HU Bin,et al
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| Abstract: |
| 【Objective】To provide a new perspective on the mechanism of IBDV-induced apoptosis,this study investigated the changes in 5-mC level in the promoter regions of apoptosis related genes after IBDV infection.【Method】Twenty one days old broilers without maternal IBDV antibodies were randomly divided into two groups of control and IBDV.Broilers in IBDV group were inoculated with IBDV CJ801 at the dosage of 0.1 mL through the oculonasal pathways,and broilers in the control group were infected with phosphate buffered saline (PBS) similarly. Bursae was collected on day 5 post inoculation.HE staining was conducted for histological analysis.The mRNA expression of apoptosis related genes was examined using real-time PCR.MeDIP PCR was used to analyze the methylation level in the promoter regions of the relative genes.【Result】Compared with control,IBDV induced tissue edema,follicle area reduction,and nuclear condensation in the bursae.The mRNA expressions of Caspase-8 and Caspase-3 were significantly decreased (P＜0.01),but Bcl-2 mRNA expression was significantly increased (P＜0.01).MeDIP PCR analysis showed that the methylation level in the promoter region of Caspase-3 was increased (P＜0.05),and the methylation level in the promoter region of Bcl-2 was remarkably decreased (P＜0.01).【Conclusion】The epigenetic regulation may be involved in the changes of expression of apoptosis-related genes during IBDV infection. |
| Key words: IBDV apoptosis apoptotic gene DNA methylation |
