短期使用达格列净加重CCl_(4)诱导的急性肝损伤

Short-term medication of dapagliflozin exacerbates CCl_(4)induced acute liver injury

目的探究钠-葡萄糖共转运体2抑制剂达格列净(dapagliflozin,Dapa)在急性肝损伤中的作用及其机制。方法8周龄C57BL6/J小鼠给予单次腹腔注射CCl_(4)诱导急性肝损伤,实验组注射前24 h和前2 h分别给予5 mg/kg Dapa预防性灌胃给药,对照组给予等体积溶剂灌胃,造模24 h后麻醉处死,采血并取肝脏。HE染色、血浆生化检测、RTq PCR及Western blotting方法检测肝脏损伤严重程度与巨噬细胞极化相关基因表达情况。结果CCl_(4)造模组小鼠肝内炎症细胞浸润,肝功、肾功能指标明显恶化,Dapa干预后肝肾功损害进一步加重。CCl_(4)能够促进肝脏巨噬细胞M1型与纤维化相关基因转录上调,同时降低M2型与抗氧化应激相关基因转录,并且能被Dapa进一步抑制。此外,Dapa干预后肝脏Arg1蛋白表达降低,SGLT2蛋白表达升高,而NF-κB通路蛋白无明显改变,提示Dapa可能通过直接作用而影响肝内能量代谢稳态,加重CCl_(4)诱导的急性肝损伤。结论Dapa可加重肝损伤急性期的肝肾功能损害,抑制巨噬细胞M2型极化,加剧CCl_(4)诱导的肝内氧化应激与炎症损伤。

Objective To investigate the role and mechanism of dapagliflozin(Dapa),a sodium glucose cotransporter 2 inhibitor,in acute liver injury.Methods Eight-week-old C57BL6/J mice were given a single intraperitoneal injection of CCl_(4)to induce acute liver injury.The mice were preventively given 5 mg/kg Dapa by gavage 24 h and 2 h before CCl_(4)injection,while those in the control group were given an equal volume of solvent gavage.After 24 h,the mice were anesthetized and sacrificed.H&E staining,plasma biochemistry,RT-q PCR,and Western blotting were used to detect the severity of liver injury and the expressions of macrophage-related genes.Results In the CCl_(4)group,hepatic infiltration of inflammatory cells increased,and liver and renal functions significantly deteriorated,which was further aggravated by Dapa.CCl_(4)could promote the expressions of M1 macrophages and fibrosis-related genes in the liver,but reduce those of M2 and antioxidant-related genes,and the latter was further inhibited by Dapa.In addition,the protein expression of arginase 1 decreased and that of SGLT2 increased after Dapa intervention,while NF-κB pathway did not change significantly,suggesting that Dapa might directly affect the energy metabolism homeostasis in the liver and aggravate acute liver injury induced by CCl_(4).Conclusion Dapa can exacerbate hepatic and renal damage in acute stage of liver injury,inhibit macrophages M2 polarization,and aggravate oxidative stress and inflammatory injury induced by CCl_(4).