IHH过表达对鸡原代软骨细胞增殖和分化的影响

doi:10.11843/j.issn.0366-6964.2023.11.013

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (11): 4569-4576.doi: 10.11843/j.issn.0366-6964.2023.11.013

IHH过表达对鸡原代软骨细胞增殖和分化的影响

金四华, 贾富民, 何培莉, 贾羽晴, 税斐, 刘旭玲, 刘星, 王新, 徐嫚, 耿照玉*

安徽农业大学动物科技学院, 合肥 230036

收稿日期:2022-10-08 出版日期:2023-11-23 发布日期:2023-11-26
通讯作者: 耿照玉,主要从事家禽遗传育种研究,E-mail:gzy@ahau.edu.cn
作者简介:金四华(1984-),安徽安庆人,博士,副教授,主要从事家禽分子遗传育种研究,E-mail:jsh3235@126.com
基金资助:
国家自然科学基金(31802028);高校优秀青年人才支持重点项目(gxyqZD2022017);安徽省教育厅高校自然科学研究重点项目(2022AH050928);教育部产学研合作协同育人项目(221000488095409);安徽省家禽产业技术体系专项经费(AHCYJSTX-06)

Effect of IHH Gene Overexpression on Proliferation and Differentiation of Chicken Primary Chondrocytes

JIN Sihua, JIA Fumin, HE Peili, JIA Yuqing, SHUI Fei, LIU Xuling, LIU Xing, WANG Xin, XU Man, GENG Zhaoyu*

College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China

Received:2022-10-08 Online:2023-11-23 Published:2023-11-26

摘要/Abstract

摘要： 为探究印度刺猬因子(IHH)基因对鸡原代软骨细胞增殖和分化的调控作用,本研究根据IHH基因序列信息,构建真核过表达载体(OV-IHH);采用双酶切和测序进行鉴定,并将其转染鸡原代软骨细胞;采用CCK-8法检测细胞增殖,流式细胞术检测细胞周期和细胞凋亡,对软骨细胞进行成骨诱导分化至21 d,采用ALP试剂盒检测细胞ALP活性。结果表明,IHH真核过表达载体构建成功,与对照组和空载体组(OV-NC)相比,过表达组IHH基因的mRNA表达显著增加(P<0.01),过表达效率扩大2 700倍左右。过表达IHH促进细胞周期从G1期到S期的转换,从而促进增殖,抑制细胞凋亡,且ALP活性升高,促进细胞分化。综上,IHH基因显著影响鸡软骨细胞的生长发育,研究结果为进一步解析IHH基因调控鸡匍匐性状的作用机制提供理论依据。

关键词: IHH, 鸡原代软骨细胞, 过表达, 增殖, 分化

Abstract: The study aimed to explore the regulatory effect of the Indian hedgehog (IHH) gene on the proliferation and differentiation of chicken primary chondrocytes. The eukaryotic overexpression vector (OV-IHH) was constructed according to the sequence information of the IHH gene in this study. The OV-IHH vector was identified by double enzyme digestion and sequencing, and transfected into chicken primary chondrocytes. The cell proliferation was detected by CCK-8 method, and the cell cycle and apoptosis were detected by flow cytometry. The chondrocytes were induced to differentiate into osteoblasts for 21 d. The ALP activity of cells was detected by the ALP kit. The results showed that the IHH eukaryotic overexpression vector was successfully constructed. Compared with the control group and the empty vector group (OV-NC), the mRNA expression of the IHH gene in the overexpression group was significantly increased (P<0.01), and the overexpression efficiency was increased about 2 700 times. Moreover, the overexpression of IHH promoted the transformation of the cell cycle from G1 to S phase, thereby promoting proliferation and inhibiting apoptosis, increasing ALP activity, and promoting cell differentiation. In conclusion, the IHH gene significantly affects the growth and development of chicken chondrocytes, which provides a theoretical basis for further analysis of the mechanism of the IHH gene regulating the creeper trait at the cellular level.

Key words: IHH, chicken primary chondrocytes, overexpression, proliferation, differentiation

中图分类号:

S831.2

金四华, 贾富民, 何培莉, 贾羽晴, 税斐, 刘旭玲, 刘星, 王新, 徐嫚, 耿照玉. IHH过表达对鸡原代软骨细胞增殖和分化的影响[J]. 畜牧兽医学报, 2023, 54(11): 4569-4576.

JIN Sihua, JIA Fumin, HE Peili, JIA Yuqing, SHUI Fei, LIU Xuling, LIU Xing, WANG Xin, XU Man, GENG Zhaoyu. Effect of IHH Gene Overexpression on Proliferation and Differentiation of Chicken Primary Chondrocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4569-4576.

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IHH过表达对鸡原代软骨细胞增殖和分化的影响

Effect of IHH Gene Overexpression on Proliferation and Differentiation of Chicken Primary Chondrocytes

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