Page 7 - 南京医科大学学报自然科学版

第42卷第5期 南京医科大学学报（自然科学版）
2022年5月 Journal of Nanjing Medical University（Natural Sciences） ·603 ·

·基础研究·

成熟 SVFs 细胞与 Hepa1⁃6 细胞共培养模型的建立及对肝细胞

脂代谢的影响

王雨竹，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：建立小鼠成熟脂肪细胞血管基质部分（stromal vascular fraction，SVF）与小鼠肝细胞（Hepa1⁃6）共培养模型，研
究成熟脂肪细胞对肝细胞脂质堆积以及代谢的影响。方法：从小鼠皮下脂肪分离提取原代SVF细胞，经体外分化成熟后，以
Transwell小室建立成熟SVF细胞和Hepa1⁃6细胞间接共培养48 h体系。应用逆转录⁃聚合酶链反应法（RT⁃PCR法）检测SVF细
胞PPARγ和PGC⁃1α等分化相关基因及Hepa1⁃6细胞CD36、FATP2、GPAT1等脂代谢相关基因的表达。酶法检测经共培养后的
Hepa1⁃6细胞甘油三酯水平。结果：小鼠原代SVF细胞经诱导成熟后，出现明显较大脂滴；成熟SVF细胞中PPARγ和PGC⁃1α等
分化标志物较未分化 SVF 细胞显著升高；Hepa1⁃6 细胞甘油三酯水平明显高于未共培养组；Hepa1⁃6 细胞 GPAT1、CD36 基因
表达水平明显升高。结论：已成功建立成熟脂肪细胞与 Hepa1⁃6 细胞 Transwell 共培养模型。肝细胞与成熟脂肪细胞共培
养可以诱导其脂质堆积增加，该过程可能通过上调肝细胞中 CD36 和 GPAT1 的表达水平增加脂肪酸的吸收和甘油三酯的
合成通路实现。
［关键词］ SVF；肝细胞；细胞共培养；脂代谢
［中图分类号］ R329.25 ［文献标志码］ A ［文章编号］ 1007⁃4368（2022）05⁃603⁃07
doi：10.7655/NYDXBNS20220501

Establishment of co⁃culture of mature SVFs with Hepa1⁃6 as a model for studying effects
on lipid metabolism of hepatocytes

WANG Yuzhu，ZHANG Xu，LI Zhong *
Key Laboratory of Rare Metabolic Disease，Department of Molecular Biology and Biochemistry，The Key Laboratory of
Human Functional Genomics of Jiangsu Province，Nanjing Medical University，Nanjing 211166，China

［Abstract］ Objective：This study aims to establish a co⁃culture system of matured stromal vascular fraction（SVF）with Hepa1⁃6
and study the effects on lipid metabolism of hepatocytes. Method：Primary SVF cells were isolated from mice subcutaneous fat and
induced to matured SVF cells with differentiation medium. Matured SVF cells were co⁃cultured with Hepa1⁃6 by using Transwell
system for 2 days. RT⁃PCR was applied to detect the expression of differentiation biomarker such as PPARγ and PGC⁃1α in SVF cells
and lipid metabolism related genes such as CD36，FATP2 and GPAT1 in Hepa1⁃6. Cellular triglyceride level of co⁃cultured Hepa1⁃6
was detected by GPO Enzyme Method. Results：Isolated primary SVF cells were successfully differentiated to mature SVF cells with
significantly large lipid droplets after treated with adipocytes differentiation medium for 6 days. The expression level of PPARγ and
PGC⁃1α of SVF cells was significantly increased. The cellular triglyceride level of co⁃cultured Hepa1⁃6 was significantly higher than
that of control group accompanied with increased expression level of GPAT1 and CD36. Conclusion：The co ⁃ cultured system of
matured SVF cells with Hepa1⁃6 in Transwell insert has been successfully established. Co⁃culture with mature SVF cells induces lipid
accumulation in Hepa1⁃6. This process may be maintained by the elevated expression level of CD36 and GPAT1，increasing fatty acid
absorption and triglyceride synthesis.
［Key words］ SVF；hepatocytes；co⁃culture，lipid metabolism
［J Nanjing Med Univ，2022，42（05）：603⁃609］
［基金项目］ 国家自然科学基金面上项目（91957115）；南京医科大学科技发展基金⁃面上项目（2016NJMU004）
∗
通信作者（Corresponding author），E⁃mail：lizhong@njmu.edu.cn