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Special Issue:
动物科学合辑Animal Science
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| HBP1 inhibits chicken preadipocyte differentiation by activating the STAT3 signaling via directly enhancing JAK2 expression |
CHEN Hong-yan1, 2, 3*, CHENG Bo-han1, 2, 3*, MA Yan-yan1, 2, 3, ZHANG Qi1, 2, 3, LENG Li1, 2, 3，WANG Shou-zhi1, 2, 3, LI Hui1, 2, 3
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1 Key Laboratory of Chicken Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Harbin 150030, P. R. China
2 Key Laboratory of Animal Genetics, Breeding and Reproduction, Education Department of Heilongjiang Province, Harbin 150030, P. R. China
3 College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, P. R. China
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摘要
本研究以东北农业大学肉鸡高、低腹脂双向选择品系1-7周龄鸡只和永生化鸡前脂肪细胞（ICP2细胞）为材料，利用RT-qPCR和Western blot方法分析HBP1在脂肪组织和前脂肪细胞分化过程中的表达模式；以稳定过表达HBP1的ICP2细胞、敲除HBP1的ICP2细胞以及各自的对照细胞为材料，利用油红O染色、RT-qPCR和Western blot检测过表达/敲除HBP1对鸡前脂肪细胞分化的影响；利用信号通路分析试剂盒筛选HBP1调控鸡前脂肪细胞分化的潜在信号通路；在稳定过表达HBP1的ICP2细胞中添加信号转导和转录激活因子3（STAT3）的化学抑制剂或转染siRNA进行功能拯救实验。结果：基因表达分析结果表明，HBP1的表达与鸡腹部脂肪沉积和前脂肪细胞分化有关。过表达HBP1抑制鸡前脂肪细胞分化（P<0.05），敲除HBP1促进鸡前脂肪细胞分化（P<0.05）。进一步研究发现，HBP1靶向激活Janus激酶2（JAK2）基因的转录来激活STAT3信号通路。功能拯救实验结果表明，STAT3信号介导了HBP1对鸡前脂肪细胞分化的调控作用。以上结果表明，HBP1通过直接上调JAK2的表达来激活STAT3信号通路，从而抑制鸡前脂肪细胞的分化。本研究阐明了HBP1在鸡前脂肪细胞分化中的基本功能，并揭示了其部分分子机制。这些发现为进一步解析鸡脂肪组织生长发育的分子遗传基础提供了新的见解。
Abstract
Obesity presents a serious threat to human health and broiler performance. The expansion of adipose tissue is mainly regulated by the differentiation of preadipocytes. The differentiation of preadipocytes is a complex biological process regulated by a variety of transcription factors and signaling pathways. Previous studies have shown that the transcription factor HMG-box protein 1 (HBP1) can regulate the differentiation of mouse 3T3-L1 preadipocytes by activating the Wnt/β-catenin signaling pathway. However, it is unclear whether HBP1 involved in chicken preadipocyte differentiation and which signaling pathways it regulates. The aim of the current study was to explore the biological function and molecular regulatory mechanism of HBP1 in the differentiation of chicken preadipocytes. The expression patterns of chicken HBP1 in abdominal adipose tissue and during preadipocyte differentiation were analyzed by RT-qPCR and Western blot. The preadipocyte stably overexpressing HBP1 or knockout HBP1 and their control cell line were used to analyze the effect of HBP1 on preadipocyte differentiation by oil red O staining, RT-qPCR and Western blot. Cignal 45-Pathway Reporter Array was used to screen the signal pathways that HBP1 regulates in the differentiation of chicken preadipocytes. Chemical inhibitor and siRNA for signal transducer and activator of transcription 3 (STAT3) were used to analyze the effect of STAT3 on preadipocyte differentiation. The preadipocyte stably overexpressing HBP1 was transfected by the siRNA of STAT3 or treated with a chemical inhibitor of STAT3 for the rescue experiment. The results of gene expression analysis showed that the expression of HBP1 was related to abdominal fat deposition and preadipocyte differentiation in chickens. The results of function gain and loss experiments indicated that overexpression/knockout of HBP1 in chicken preadipocytes could inhibit/promote (P<0.05) lipid droplet deposition and the expression of adipogenesis-related genes. Mechanismlly, HBP1 activates (P<0.05) the signal transducer and activator of transcription 3 (STAT3) signaling pathway by targeting janus kinase 2 (JAK2) transcription. The results of functional rescue experiments indicated that STAT3 signaling mediated the regulation of HBP1 on chicken preadipocyte differentiation. In conclusion, HBP1 inhibits chicken preadipocyte differentiation by activating the STAT3 signaling pathway via directly enhancing JAK2 expression. Our findings provided new insights for further analysis of the molecular genetic basis of chicken adipose tissue growth and development.
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Received: 07 June 2021
Accepted: 19 January 2022
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| Fund:
This research was supported by the China Agriculture Research System of MOF and MARA of China (CARS-41).
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| About author: CHEN Hong-yan, E-mail: chen_hongyan@neau.edu.cn; CHENG Bo-han, E-mail: bhcheng@neau.edu.cn; Correspondence LI Hui, Tel/Fax: +86-451-55191516, E-mail: lihui@neau.edu.cn
* These authors contributed equally to this study. |
Cite this article:
CHEN Hong-yan, CHENG Bo-han, MA Yan-yan, ZHANG Qi, LENG Li, WANG Shou-zhi, LI Hui.
2022.
HBP1 inhibits chicken preadipocyte differentiation by activating the STAT3 signaling via directly enhancing JAK2 expression. Journal of Integrative Agriculture, 21(6): 1740-1754.
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