SGLT2 inhibitor empagliflozin downregulates miRNA-34a-5p and targets GREM2 to inactivate hepatic stellate cells and ameliorate non-alcoholic fatty liver disease-associated fibrosis.
SGLT2 inhibitor empagliflozin downregulates miRNA-34a-5p and targets GREM2 to inactivate hepatic stellate cells and ameliorate non-alcoholic fatty liver disease-associated fibrosis.
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DOI：
10.1016/j.metabol.2023.155657
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发表时间：
2023-07
期刊：
影响因子：
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通讯作者：
Yunfeng Shen;Lidan Cheng;Minxuan Xu;Wei Wang;Zhiping Wan;Haixia Xiong;Wanrong Guo;Meng-yin Cai;Fen Xu
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作者：
Yunfeng Shen;Lidan Cheng;Minxuan Xu;Wei Wang;Zhiping Wan;Haixia Xiong;Wanrong Guo;Meng-yin Cai;Fen Xu
肝星状细胞（hepatic stellate cells，HSC）的活化是肝纤维化的中心事件，提示非酒精性脂肪性肝病（nonalcoholic fatty liver disease，NAFLD）的严重程度。microRNAs（miRNAs）参与了这一过程。钠-葡萄糖协同转运蛋白2抑制剂（SGLT 2 i）治疗可加重2型糖尿病和NAFLD患者的肝纤维化;然而，SGLT 2 i通过调节miRNAs在改善NAFLD患者肝纤维化中的作用尚不清楚。方法和结果我们监测了两种NAFLD模型肝脏中NAFLD相关miRNAs的表达，并观察到miR-34 a-5 p的高表达。miR-34 a-5 p在小鼠原代肝非实质细胞和LX-2 HSC中高表达，且该miRNA与NAFLD模型中丙氨酸转氨酶水平呈正相关。miR-34 a-5 p的过表达增强LX-2的活化，而其抑制通过调节TGFβ信号通路阻止HSC活化。SGLT 2 i恩格列净显著下调miR-34 a-5 p，抑制TGFβ信号通路，并改善NAFLD模型中的肝纤维化。随后，通过数据库预测和双荧光素酶报告基因测定，GREM 2被鉴定为miR-34 a-5 p的直接靶点。在LX-2 HSC中，miR-34 a-5 p模拟物和抑制剂分别直接下调和上调GREM 2。过表达GREM 2使TGFβ通路失活，而GREM 2敲低使其活化。此外，恩格列净上调NAFLD模型中的GREM 2表达。在蛋氨酸和胆碱缺乏饮食喂养的ob/ob小鼠中，纤维化模型，恩格列净下调miR-34 a-5 p和上调Grem 2以改善肝纤维化。结论恩格列净通过下调miR-34 a-5 p和靶向Grem 2抑制HSC中的TGFβ通路来改善NAFLD相关的纤维化。
Background and rationaleActivation of hepatic stellate cells (HSCs), the central event of fibrosis, indicates the severe stage of non-alcoholic fatty liver disease (NAFLD). MicroRNAs (miRNAs) participate in this process. Treatment with a sodium-glucose cotransporter 2 inhibitor (SGLT2i) alleviates liver fibrosis in patients with type 2 diabetes and NAFLD; however, the role of SGLT2i in ameliorating liver fibrosis in NAFLD by regulating miRNAs remains unclear.Approach and resultsWe monitored the expression of NAFLD-associated miRNAs in the livers of two NAFLD models and observed high expression of miR-34a-5p. miR-34a-5p was highly expressed in mouse primary liver non-parenchymal cells and LX-2 HSCs, and this miRNA was positively correlated with alanine transaminase levels in NAFLD models. Overexpression of miR-34a-5p enhanced LX-2 activation, whereas its inhibition prevented HSCs activation by regulating the TGFβ signaling pathway. The SGLT2i empagliflozin significantly downregulated miR-34a-5p, inhibited the TGFβ signaling pathway, and ameliorated hepatic fibrosis in NAFLD models. Subsequently,GREM2was identified as a direct target of miR-34a-5p through database prediction and a dual-luciferase reporter assay. In LX-2 HSCs, the miR-34a-5p mimic and inhibitor directly downregulated and upregulatedGREM2, respectively. OverexpressingGREM2inactivated the TGFβ pathway whereasGREM2knockdown activated it. Additionally, empagliflozin upregulatedGrem2expression in NAFLD models. In methionine- and choline-deficient diet-fed ob/ob mice, a fibrosis model, empagliflozin downregulated miR-34a-5p and upregulatedGrem2to improve liver fibrosis.ConclusionsEmpagliflozin ameliorates NAFLD-associated fibrosis by downregulating miR-34a-5p and targetingGREM2to inhibit the TGFβ pathway in HSCs.