重组鸡GSTA3蛋白对福美双诱导胫骨软骨发育不良肉鸡红细胞免疫相关基因转录的影响

doi:10.11843/j.issn.0366-6964.2018.04.019

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (4): 811-817.doi: 10.11843/j.issn.0366-6964.2018.04.019

重组鸡GSTA3蛋白对福美双诱导胫骨软骨发育不良肉鸡红细胞免疫相关基因转录的影响

贾发杰¹, 牛胜¹, 张宁¹, 李欣¹, 宁官保¹, 张鼎¹, 李宏全¹, 马海利¹, 郝卫芳², 高文伟¹, 赵宇军¹, 高诗敏¹, 李桂兰¹, 李建慧¹, 闫芳¹, 高荣琨¹, 田文霞^1*

1. 山西农业大学动物科技学院, 太谷 030801;
2. 太原市动物疫病预防控制中心, 太原 030024

收稿日期:2017-10-09 出版日期:2018-04-23 发布日期:2018-04-23
通讯作者: 田文霞,E-mail:wenxiatian@126.com
作者简介:贾发杰(1989-),男,河南商丘人,硕士生,主要从事动物传染病发病与免疫机制研究,E-mail:2251651836@qq.com。
基金资助:
国家重点研发计划（2016YFD0500800）；山西省回国留学人员科研资助项目（2017-073）；晋中市重点科技创新平台（P171002-3）；山西省科技攻关项目（20130311027-3）；国家自然科学基金（31072179）；山西省自然基金（2014011028-1）

The Effection of Recombinant Protein GSTA3 on the Transcription of Erythrocytes Immune Related Genes in Tibial Dyschondroplasia Broilers Induced by Thiram

JIA Fa-jie¹, NIU Sheng¹, ZHANG Ning¹, LI Xin¹, NING Guan-bao¹, ZHANG Ding¹, LI Hong-quan¹, MA Hai-li¹, HAO Wei-fang², GAO Wen-wei¹, ZHAO Yu-jun¹, GAO Shi-min¹, LI Gui-lan¹, LI Jian-hui¹, YAN Fang¹, GAO Rong-kun¹, TIAN Wen-xia^1*

1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China;
2. Taiyuan Center for Disease Control and Prevention, Taiyuan 030024, China

Received:2017-10-09 Online:2018-04-23 Published:2018-04-23

摘要/Abstract

摘要：

为研究胫骨软骨发育不良（TD）肉鸡红细胞免疫相关基因转录水平的变化及重组鸡谷胱甘肽S-转移酶A3（chGSTA3）蛋白对TD肉鸡红细胞免疫相关基因转录的影响，将120只肉雏鸡饲喂一周后，随机分为基础日粮组（A、B、C组）和福美双处理组（D、E、F组），对福美双处理组肉鸡通过饲料中添加100 mg·kg^-1福美双诱发建立肉鸡TD模型，通过腿部肌肉注射法，对B组和E组肉雏鸡注射有活性的低剂量（20 μg·kg^-1）重组GSTA3蛋白，对C组和F组肉雏鸡注射高剂量（50 μg·kg^-1）重组GSTA3蛋白，对A和D组的肉雏鸡注射等体积的磷酸盐缓冲液（PBS）；使用Real-time PCR对肉鸡红细胞中免疫基因的信使RNA（mRNA）转录水平进行检测。结果显示，Toll样受体（TLR）2、TLR3、TLR4、TLR5、TLR7、TLR15、髓样分化因子88（MyD88）、主要组织相容性复合体（MHC）Ⅱ、核苷酸结合寡聚化结构域样受体家族中的C5型受体（NLRC5）、肿瘤坏死因子受体相关因子6（TRAF6）、白细胞介素-7（IL-7）基因可以在鸡红细胞转录；福美双处理组D与磷酸缓冲盐溶液（PBS）处理组A相对比，TLR2、TLR3、TLR4、TLR5、TLR7、TLR15、MyD88、TRAF6、MHCⅡ和NLRC5在试验的第4天显著升高，IL-7显著降低（P<0.05），在试验的第15天TD肉鸡红细胞中TLR2、TLR4、MyD88和NLRC5显著下调（P<0.05），IL-7、TLR3、TLR5、TLR7、TLR15、MHCⅡ、TRAF6变化不明显（P>0.05）；E组和F组与D组相比，在试验第4天，除E组TLR4、TLR5和MyD88外，其余基因都有显著性变化（P<0.05）；在第15天，E组中TLR4、TLR5、MyD88和NLRC5，及F组中TLR3、TLR5、MyD88、TRAF6和IL-7变化不显著，其他基因都有显著性差异（P<0.05）。鸡红细胞可能通过免疫相关基因TLR2、TLR3、TLR4、TLR5、TLR7、TLR15、IL-7、MyD88、TRAF6、MHCⅡ和NLRC5转录水平的变化，在早期诱导TD发生，后期减轻肉鸡TD症状；GSTA3能够对福美双诱导肉鸡TD的作用产生影响。

Abstract:

The objective of this study was to evaluate the transcriptional changes of immune related genes and the effection of recombinant chicken glutathione S-transferase A3 (chGSTA3) protein on the transcription of immune related genes in erythrocytes of Tibial Dyschondroplasia (TD) broiler chickens. Therefore, the model of TD broiler chickens was established, in which 120 broiler chickens were randomly broken up into 6 groups; group A, B and C (fed with basal diet), group D, E and F (fed with basal diet containing 100 mg·kg^-1 Thiram) after one week normal feeding. The broiler chickens of group B and E were treated with chGSTA3 protein by intramuscular injection (low dose:20 μg·kg^-1) and the broilers of group C and F were treated intra-muscularly with chGSTA3 protein (high dose:50 μg·kg^-1), while groups A and D received the same dosage of phosphate buffered solution (PBS). We used Real-time PCR to investigate the mRNA expression of the immune-related genes in chicken erythrocytes. The transcripts for TLR2, TLR3, TLR4, TLR5, TLR7, TLR15, myeloid differentiation factor 88 (MyD88), major histocompatibility complex (MHC) class Ⅱ, Nucleotidebinding oligomerization domain receptor caspase recruitment domain 5 (NLRC5), TNF receptor-associated factor 6 (TRAF6) and Interleukin (IL-7) were constitutively expressed in erythrocytes of healthy broiler chickens. Contrasted with group A with phosphate buffered solution (PBS) treatment, the mRNA transcription level of TLR2, TLR3, TLR4, TLR5, TLR7, TLR15, MyD88, MHCⅡ, NLRC5, TRAF6 of Thiram-treated chicken erythrocytes in group D on the 4^th day were significantly upregulated (P<0.05), whereas IL-7 was significantly downregulated. We observed that the transcripts of immune-related genes:TLR2, TLR4, MyD88 and NLRC5 in the chicken erythrocytes of group D were significantly downregulated than that in group A (P<0.05), but IL-7, TLR3, TLR5, TLR7, TLR15, MHCⅡ and TRAF6 were insignificant (P>0.05) on the 15^th day. The changes of above mentioned genes mRNA expression in group E and F were significant except TLR4, TLR5, MyD88 in group E as compared with group D on the 4^th day. The changes of TLR4, TLR5, MyD88, NLRC5 genes in group E and TLR3, TLR5, MyD88, TRAF6, IL-7 genes in group F were insignificant, whereas the others were significant compared with group D on the 15^th day. Broiler chicken erythrocytes could induce the occurrence of TD in the early stage and alleviate TD symptoms in the late stage by changing the mRNA expression of IL-7, TLR2, TLR3, TLR4, TLR5, TLR7, TLR15, MyD88, MHCⅡ, NLRC5, TRAF6. The recombinant chicken glutathione S-transferase A3 protein could influence TD development caused by Thiram.

中图分类号:

S852.34

贾发杰, 牛胜, 张宁, 李欣, 宁官保, 张鼎, 李宏全, 马海利, 郝卫芳, 高文伟, 赵宇军, 高诗敏, 李桂兰, 李建慧, 闫芳, 高荣琨, 田文霞. 重组鸡GSTA3蛋白对福美双诱导胫骨软骨发育不良肉鸡红细胞免疫相关基因转录的影响[J]. 畜牧兽医学报, 2018, 49(4): 811-817.

JIA Fa-jie, NIU Sheng, ZHANG Ning, LI Xin, NING Guan-bao, ZHANG Ding, LI Hong-quan, MA Hai-li, HAO Wei-fang, GAO Wen-wei, ZHAO Yu-jun, GAO Shi-min, LI Gui-lan, LI Jian-hui, YAN Fang, GAO Rong-kun, TIAN Wen-xia. The Effection of Recombinant Protein GSTA3 on the Transcription of Erythrocytes Immune Related Genes in Tibial Dyschondroplasia Broilers Induced by Thiram[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(4): 811-817.

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重组鸡GSTA3蛋白对福美双诱导胫骨软骨发育不良肉鸡红细胞免疫相关基因转录的影响

The Effection of Recombinant Protein GSTA3 on the Transcription of Erythrocytes Immune Related Genes in Tibial Dyschondroplasia Broilers Induced by Thiram

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