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第43卷第8期 南京医科大学学报(自然科学版)
2023年8月 Journal of Nanjing Medical University(Natural Sciences) ·1041 ·
·基础研究·
VPS13A在3T3⁃L1脂肪细胞分化过程中的表达及调控研究
刘 瑾,黄 昀,李超普,张 桐,潘锦堃,季学涛,张 许,李 仲 *
南京医科大学罕见代谢性疾病研究重点实验室,南京医科大学生物化学与分子生物学系,江苏省人类功能基因组重点实验室,
江苏 南京 211166
[摘 要] 目的:明确囊泡分选蛋白13A(vacuolar protein sorting 13 homolog A,VPS13A)在小鼠胚胎成纤维细胞(mouse embry⁃
onic fibroblast cells,3T3⁃L1)分化过程中的表达水平;探索VPS13A对3T3⁃L1细胞的影响。方法:利用Western blot及Q⁃PCR法
检测3T3⁃L1细胞在分化过程中VPS13A的表达,利用CRISPR/Cas9系统构建VPS13A稳定敲降细胞系,Western blot和油红染色
检测VPS13A敲降后对3T3⁃L1细胞分化的影响。结果:VPS13A的表达在3T3⁃L1分化过程的早期降低,在分化的后期升高。敲
降VPS13A后,3T3⁃L1细胞中脂滴增多,参与调控脂肪细胞分化基因的表达水平升高。结论:在3T3⁃L1细胞中VPS13A的表达
水平在分化过程中被调控,敲降VPS13A可以促进3T3⁃L1细胞的分化成熟。
[关键词] VPS13A;3T3⁃L1细胞;脂滴;脂肪细胞分化
[中图分类号] R329.26 [文献标志码] A [文章编号] 1007⁃4368(2023)08⁃1041⁃06
doi:10.7655/NYDXBNS20230801
Expression and regulation of VPS13A in 3T3⁃L1 adipocyte differentiation
*
LIU Jin,HUANG Yun,LI Chaopu,ZHANG Tong,PAN Jinkun,JI Xuetao,ZHANG Xu,LI Zhong
Key Laboratory of Rare Metabolic Disease,Department of Molecular Biology and Biochemistry,Nanjing Medical
University,the Key Laboratory of Human Functional Genomics of Jiangsu Province,Nanjing 211166,China
[Abstract] Objective:To determine the expression level of vacuolar protein sorting 13 homolog A(VPS13A)during cell
differentiation of 3T3⁃L1 and to explore the effects of VPS13A on 3T3⁃L1 cells. Methods:Western blot and Q⁃PCR were used to detect
the expression changes of VPS13A during the differentiation of 3T3 ⁃ L1 cells,and CRISPR(clustered regularly interspaced short
palindromic repeats)/Cas9(CRISPR associated with 9)system to establish stable VPS13A knowdown cell lines. Finally,the effect of
VPS13A knockdown on 3T3⁃L1 cell differentiation was detected by Western blot and Oil red staining. Results:The expression of
VPS13A was downregulated in the early stage and upregulated in the later stage of 3T3⁃L1 cell differentiation. Lipid droplets and the
expression levels of genes involved in differentiation were increased in VPS13A knockdown 3T3 ⁃ L1 cells. Conclusion:VPS13A
expression is modulated by cell differentiation status in 3T3⁃L1 cells. Knockdown of VPS13A enhances the differentiation of 3T3⁃L1
cells.
[Key words] VPS13A;3T3⁃L1 cell;lipid droplets;adipocytes differentiation
[J Nanjing Med Univ,2023,43(08):1041⁃1046]
世界卫生组织发布的最新数据表明全球肥胖 引起的。白色脂肪组织通过增加脂肪细胞的大小
发病率越来越高。全球肥胖患病率的大幅增加一 或促进脂肪前体细胞的分化增加脂肪细胞数目而
[1]
方面可能是由于营养摄入过多以及运动量不足引 扩张,进一步引起肥胖症的发生 。脂肪组织在机
[2]
起的,另一方面也可能是由于遗传和生物等因素所 体能量代谢过程中起着重要作用 ,相关研究表明
肥胖与 2 型糖尿病、高血压等疾病之间具有明显相
[基金项目] 国家自然科学基金(32130050,3220110126);南
[3]
京医科大学科技发展基金(2016NJMU004) 关性 。因此对脂肪细胞分化机制的深入研究可
∗ 以为肥胖的防治提供重要的理论基础及潜在的靶
通信作者(Corresponding author),E⁃mail:lizhong@njmu.edu.cn