BACKGROUND The Tibial dyschondroplasia (TD) in fast-growing chickens is mainly caused by improper blood circulation. The exact mechanism underlying angiogenesis and vascularization in tibial growth plate of broiler chickens remains unclear. Therefore, this research attempts to study genes involved in the regulation of angiogenesis in chicken red blood cells. Twenty-four broiler chickens were allotted into a control and thiram (Tetramethyl thiuram disulfide) group. Blood samples were collected on day 2, 6 (8- and 14-days old chickens) and 15 (23 days old chickens). RESULTS Histopathology and hematoxylin and eosin (H&E) results showed that angiogenesis decreased on the 6th day of the experiment but started to recover on the 15th day of the experiment. Immunohistochemistry (IHC) results confirmed the expressions of integrin alpha-v precursor (ITGAV) and clusterin precursor (CLU). Transcriptome sequencing analysis evaluated 293 differentially expressed genes (DEGs), of which 103 up-regulated genes and 190 down-regulated genes were enriched in the pathways of neuroactive ligand receptor interaction, mitogen-activated protein kinase (MAPK), ribosome, regulation of actin cytoskeleton, focal adhesion, natural killer cell mediated cytotoxicity and the notch signalling pathways. DEGs (n = 20) related to angiogenesis of chicken erythrocytes in the enriched pathways were thromboxane A2 receptor (TBXA2R), interleukin-1 receptor type 1 precursor (IL1R1), ribosomal protein L17 (RPL17), integrin beta-3 precursor (ITGB3), ITGAV, integrin beta-2 precursor (ITGB2), ras-related C3 botulinum toxin substrate 2 (RAC2), integrin alpha-2 (ITGA2), IQ motif containing GTPase activating protein 2 (IQGAP2), ARF GTPase-activating protein (GIT1), proto-oncogene vav (VAV1), integrin alpha-IIb-like (ITGA5), ras-related protein Rap-1b precursor (RAP1B), tyrosine protein kinase Fyn-like (FYN), tyrosine-protein phosphatase non-receptor type 11 (PTPN11), protein patched homolog 1 (PTCH1), nuclear receptor corepressor 2 (NCOR2) and mastermind like protein 3 (MAML3) selected for further confirmation with qPCR. However, commonly DEGs were sarcoplasmic/endoplasmic reticulum calcium ATPase 3 (ATP2A3), ubiquitin-conjugating enzyme E2 R2 (UBE2R2), centriole cilia and spindle-associated protein (CCSAP), coagulation factor XIII A chain protein (F13A1), shroom 2 isoform X6 (SHROOM2), ras GTPase-activating protein 3 (RASA3) and CLU. CONCLUSION We have found potential therapeutic genes concerned to erythrocytes and blood regulation, which regulated the angiogenesis in thiram induced TD chickens. This study also revealed the potential functions of erythrocytes. 1. Tibial dyschondroplasia (TD) in chickens were more on day 6, which started recovering on day 15. 2. The enriched pathway observed in TD chickens on day 6 was ribosome pathway, on day 15 were regulation of actin cytoskeleton and focal adhesion pathway. 3. The genes involved in the ribosome pathways was ribosomal protein L17 (RPL17). regulation of actin cytoskeleton pathway were Ras-related C3 botulinum toxin substrate 2 (RAC2), Ras-related protein Rap-1b precursor (RAP1B), ARF GTPase-activating protein (GIT1), IQ motif containing GTPase activating protein 2 (IQGAP2), Integrin alpha-v precursor (ITGAV), Integrin alpha-2 (ITGA2), Integrin beta-2 precursor (ITGB2), Integrin beta-3 precursor (ITGB3), Integrin alpha-IIb-like (ITGA5). Focal adhesion Proto-oncogene vav (Vav-like), Tyrosine-protein kinase Fyn-like (FYN).

中文翻译：

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基于转录组的筛查法筛选肉鸡在不同阶段由锡拉姆诱导的胫骨病变中的细胞内途径和血管生成相关基因。

背景技术快速生长的鸡中的胫骨软骨发育不良（TD）主要是由血液循环不当引起的。肉鸡胫骨生长板中血管生成和血管形成的确切机制尚不清楚。因此，本研究试图研究参与调节鸡红细胞中血管生成的基因。将二十四只肉鸡分为对照组和硫柳（四甲基秋兰姆二硫化物）组。在第2、6（8和14天大的鸡）和15（23天大的鸡）收集血样。结果组织病理学，苏木精和曙红（H＆E）结果表明，血管生成在实验的第6天减少，但在实验的第15天开始恢复。免疫组织化学（IHC）结果证实了整联蛋白α-v前体（ITGAV）和簇蛋白前体（CLU）的表达。转录组测序分析评估了293个差异表达基因（DEG），其中103个上调基因和190个下调基因丰富了神经活性配体受体相互作用，促分裂原激活蛋白激酶（MAPK），核糖体，肌动蛋白调节的途径细胞骨架，粘着斑，自然杀伤细胞介导的细胞毒性和Notch信号通路。与富集途径中的鸡红细胞血管生成有关的DEG（n = 20）是血栓烷A2受体（TBXA2R），白细胞介素1受体1型前体（IL1R1），核糖体蛋白L17（RPL17），整联蛋白beta-3前体（ITGB3） ，ITGAV，整联蛋白beta-2前体（ITGB2），与ras相关的C3肉毒毒素底物2（RAC2），整联蛋白α-2（ITGA2），包含GTPase激活蛋白2（IQGAP2）的IQ基序，ARF GTPase激活蛋白（GIT1），原癌基因vav（VAV1），整联蛋白alpha -IIb样（ITGA5），ras相关蛋白Rap-1b前体（RAP1B），酪氨酸蛋白激酶Fyn样（FYN），酪氨酸蛋白磷酸酶非受体11型（PTPN11），蛋白修补同源物1（PTCH1） ，核受体corepressor 2（NCOR2）和mastermind like protein 3（MAML3）进行qPCR进一步确认。然而，常见的DEGs是肌浆/内质网钙ATPase 3（ATP2A3），泛素结合酶E2 R2（UBE2R2），纤毛纤毛和纺锤体相关蛋白（CCSAP），凝血因子XIII A链蛋白（F13A1），shroom 2亚型。 X6（SHROOM2），ras GTP酶激活蛋白3（RASA3）和CLU。结论我们发现了潜在的治疗基因，与红细胞和血液调节有关，这些基因调节了硫柳胺诱导的TD鸡的血管生成。这项研究还揭示了红细胞的潜在功能。1.鸡的胫骨软骨发育不良（TD）在第6天开始增多，在第15天开始恢复。2.在第6天在TD鸡中观察到的富集途径是核糖体途径，在第15天是肌动蛋白细胞骨架和粘着斑粘附途径的调节。3.核糖体途径中涉及的基因是核糖体蛋白L17（RPL17）。肌动蛋白细胞骨架途径的调控包括Ras相关的C3肉毒杆菌毒素底物2（RAC2），Ras相关的蛋白Rap-1b前体（RAP1B），ARF GTPase激活蛋白（GIT1），包含GTPase激活蛋白2的IQ基序（IQGAP2），整合素α-v前体（ITGAV），整合素α-2（ITGA2），整合素β-2前体（ITGB2），整合素β-3前体（ITGB3），整合素α-IIb样（ITGA5）。局灶粘附原癌基因vav（Vav样），酪氨酸蛋白激酶Fyn样（FYN）。