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南京医科大学学报（自然科学版） 第44卷第11期
·1510 · Journal of Nanjing Medical University（Natural Sciences） 2024年11月
·基础研究·
miR⁃199a⁃5p在肝脏组织中的表达及其作用的初步研究
潘锦堃，李超普，张 许，季学涛，薛 瑶，李 仲 *
南京医科大学罕见代谢性疾病研究重点实验室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：探究不同营养状态下miR⁃199a⁃5p在肝脏中的表达水平，以及其对肝细胞甘油三酯（triglyceride，TAG）含量的
影响及其潜在机制。方法：RT⁃qPCR检测高脂饮食小鼠肝脏组织中miR⁃199a⁃5p的表达；分别使用miR⁃199a⁃5p模拟物、抑制
剂、阴性对照和pcDH⁃CD36⁃flag质粒转染Hepa1⁃6和AML12细胞，通过RT⁃qPCR和Western blot检测脂代谢相关标志物表达变
化，采用试剂盒检测TAG含量；通过miRDB（microRNA Target Prediction Database）预测miR⁃199a⁃5p的靶基因并通过双荧光素
酶报告实验进行验证。结果：高脂饮食和饥饿状态下C57BL/6J小鼠肝脏中miR⁃199a⁃5p的表达水平升高；过表达miR⁃199a⁃5p
能够降低肝细胞内TAG，而miR⁃199a⁃5p抑制剂会增加细胞内TAG含量；miR⁃199a⁃5p通过作用于脂肪酸转位酶CD36基因的
3′非翻译区（3′ untranslated region，3′ UTR）降低其蛋白表达。结论：在肝脏脂质积累过程中，miR⁃199a⁃5p的表达水平增加，并
且miR⁃199a⁃5p可能通过靶向CD36降低肝细胞内TAG含量。
［关键词］ miR⁃199a⁃5p；肝脏；甘油三酯；CD36
［中图分类号］ R575 ［文献标志码］ A ［文章编号］ 1007⁃4368（2024）11⁃1510⁃07
doi：10.7655/NYDXBNSN240674
The expression and role of miR⁃199a⁃5p in the liver
PAN Jinkun，LI Chaopu，ZHANG Xu，JI Xuetao，XUE Yao，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 investigate the expression levels of miR⁃199a⁃5p in the liver under various nutritional conditions and its
effects on hepatic triacylglyceride（TAG）content，as well as the underlying mechanisms. Methods：RT⁃qPCR was used to detect the
expression levels of miR⁃199a⁃5p in liver tissues of high fat diet mice. Hepa1⁃6 and AML12 cells were transfected with miR⁃199a⁃5p
mimics，inhibitors，negative controls and pcDH⁃CD36⁃flag plasmid，respectively. The changes in biomarker expression related to lipid
metabolism were detected by RT⁃qPCR and Western blot，and TAG content was detected by kit. The target gene of miR⁃199a⁃5p was
predicted by microRNA Target Prediction Database（miRDB）and verified by the dual luciferase reporting assay. Results：The
expression levels of miR ⁃ 199a ⁃ 5p in the liver of C57BL/6J mice were elevated under high ⁃ fat diet and fasting conditions.
Overexpression of miR⁃199a⁃5p reduced TAG levels in hepatocytes，while inhibition of miR⁃199a⁃5p increased intracellular TAG
content. miR⁃199a⁃5p decreased the protein expression of the fatty acid translocase CD36 by interacting with its 3′ untranslated region
（3′ UTR）. Conclusion：The expression level of miR⁃199a⁃5p increases during hepatic lipid accumulation，and miR⁃199a⁃5p may
reduce TAG content in hepatocytes by targeting CD36.
［Key words］ miR⁃199a⁃5p；liver；triacylglyceride；CD36
［J Nanjing Med Univ，2024，44（11）：1510⁃1516］
肝脏是体内脂代谢调控的重要器官，通过脂肪 density lipoprotein，VLDL）的分泌维持机体脂代谢稳
酸摄取、合成、氧化以及极低密度脂蛋白（very low 态 ［1-3］ 。非酒精性脂肪肝病（non⁃alcohol fatty liver dis⁃
［基金项目］ 国家自然科学基金（32130050，32201064） ease，NAFLD）是以肝细胞内脂肪过度沉积为主要特
∗ 征的临床病理综合征，是机体代谢紊乱的肝脏表
通信作者（Corresponding author），E⁃mail：lizhong@njmu.edu.cn
