Chen Hang, Huang Minshan, Zhang Dan, Wang Hui, Wang Da, Li Mengwei, Wang Xianmei, Zhu Rui, Liu Jianjun, Ma Lanqing
The First Affiliated Hospital, Yunnan Institute of Digestive Disease, Kunming Medical University, Kunming, China.
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, School of Life Sciences, Yunnan University, Kunming, China.
Front Pharmacol. 2024 Dec 17;15:1477212. doi: 10.3389/fphar.2024.1477212. eCollection 2024.
Metformin has shown benefits in treating metabolic dysfunction-associated steatotic liver disease (MASLD), but its mechanisms remain unclear. This study investigates miR-200a-5p's role in the AMPK/SERCA2b pathway to reduce liver fat accumulation and ER stress in MASLD.
A PA cell model induced by palmitic and oleic acids (2:1) was used to assess lipid accumulation via Oil Red O and Nile Red staining. mRNA levels of miR-200a-5p and lipid metabolism genes were measured with RT-PCR, and AMPK, p-AMPK, and SERCA2b protein levels were analyzed by Western blotting. The interaction between miR-200a-5p and AMPK was studied using a luciferase reporter assay. A high-fat diet-induced MASLD mouse model was used to evaluate metformin's effects on liver steatosis and lipid profiles. Serum miR-200a-5p levels were also analyzed in MASLD patients.
In the PA cell model, elevated miR-200a-5p and lipid metabolism gene mRNA levels were observed, with decreased AMPK and SERCA2b protein levels. miR-200a-5p mimic reduced AMPK and SERCA2b expression. Metformin treatment reduced liver steatosis and lipid deposition in mice, normalizing miR-200a-5p, lipid metabolism gene mRNA, and AMPK/SERCA2b protein levels. Elevated serum miR-200a-5p was detected in MASLD patients.
These findings suggest that metformin alleviates lipid deposition and ER stress in MASLD through the modulation of the AMPK/SERCA2b pathway via miR-200a-5p.
二甲双胍已显示出对治疗代谢功能障碍相关脂肪性肝病（MASLD）有益，但其机制仍不清楚。本研究探讨miR-200a-5p在AMPK/SERCA2b通路中对减少MASLD中肝脏脂肪堆积和内质网应激的作用。
使用棕榈酸和油酸（2:1）诱导的PA细胞模型，通过油红O和尼罗红染色评估脂质蓄积。用逆转录聚合酶链反应（RT-PCR）检测miR-200a-5p和脂质代谢基因的mRNA水平，并用蛋白质免疫印迹法分析AMPK、p-AMPK和SERCA2b蛋白水平。使用荧光素酶报告基因检测法研究miR-200a-5p与AMPK之间的相互作用。使用高脂饮食诱导的MASLD小鼠模型评估二甲双胍对肝脏脂肪变性和血脂谱的影响。还分析了MASLD患者血清中的miR-200a-5p水平。
在PA细胞模型中，观察到miR-200a-5p和脂质代谢基因mRNA水平升高，同时AMPK和SERCA2b蛋白水平降低。miR-200a-5p模拟物降低了AMPK和SERCA2b的表达。二甲双胍治疗减少了小鼠的肝脏脂肪变性和脂质沉积，使miR-200a-5p、脂质代谢基因mRNA和AMPK/SERCA2b蛋白水平恢复正常。在MASLD患者中检测到血清miR-200a-5p升高。
这些发现表明，二甲双胍通过miR-200a-5p调节AMPK/SERCA2b通路，减轻MASLD中的脂质沉积和内质网应激。
