Li A, Li Y Y, Wuqie Q B, Li X, Zhang H, Wang Y, Wang Y L, Zhu J J, Lin Y Q
Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization of Education Ministry, Southwest Minzu University, Chengdu 610041, China.
Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Sichuan Province, Southwest Minzu University, Chengdu 610041, China.
Anim Biosci. 2023 Jun;36(6):829-839. doi: 10.5713/ab.22.0308. Epub 2023 Jan 11.
The aim of this study was to clone the mRNA sequence of the Acyl-CoA dehydrogenase long chain (ACADL) gene of goats and explore the effect of ACADL on the differentiation of subcutaneous fat cells on this basis.
We obtained the ACADL gene of goats by cloning and used quantitative real-time polymerase chain reaction (qPCR) to detect the ACADL expression patterns of different goat tissues and subcutaneous fat cells at different lipid induction stages. In addition, we transfect intramuscular and subcutaneous adipocytes separately by constructing overexpressed ACADL vectors and synthesizing Si-ACADL; finally, we observed the changes in oil red stained cell levels under the microscope, and qPCR detected changes in mRNA levels.
The results showed goat ACADL gene expressed in sebum fat. During adipocyte differentiation, ACADL gradually increased from 0 to 24 h of culture, and decreased. Overexpression of ACADL promoted differentiation of subcutaneous adipocytes in goat and inhibited their differentiation after interference.
So, we infer ACADL may have an important role in positive regulating the differentiation process in goat subcutaneous adipocytes. This study will provide basic data for further study of the role of ACADL in goat subcutaneous adipocyte differentiation and lays the foundation for final elucidating of its molecular mechanisms in regulating subcutaneous fat deposition in goats.
本研究旨在克隆山羊酰基辅酶A脱氢酶长链(ACADL)基因的mRNA序列,并在此基础上探讨ACADL对皮下脂肪细胞分化的影响。
通过克隆获得山羊ACADL基因,采用定量实时聚合酶链反应(qPCR)检测不同山羊组织及不同脂质诱导阶段皮下脂肪细胞中ACADL的表达模式。此外,分别构建ACADL过表达载体和合成Si-ACADL转染肌内和皮下脂肪细胞;最后,在显微镜下观察油红染色细胞水平的变化,qPCR检测mRNA水平的变化。
结果显示山羊ACADL基因在皮脂中表达。在脂肪细胞分化过程中,ACADL在培养0至24小时逐渐升高,之后下降。ACADL过表达促进山羊皮下脂肪细胞分化,干扰后则抑制其分化。
因此,我们推断ACADL可能在正向调节山羊皮下脂肪细胞分化过程中发挥重要作用。本研究将为进一步研究ACADL在山羊皮下脂肪细胞分化中的作用提供基础数据,并为最终阐明其调节山羊皮下脂肪沉积的分子机制奠定基础。