Blunting p38 MAPKa and ERK1/2 activities by empagliflozin enhances the antifibrotic effect of metformin and augments its AMPK-induced NF-κB inactivation in mice intoxicated with carbon tetrachloride
Blunting p38 MAPKa and ERK1/2 activities by empagliflozin enhances the antifibrotic effect of metformin and augments its AMPK-induced NF-κB inactivation in mice intoxicated with carbon tetrachloride
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DOI：
10.1016/j.lfs.2021.120070
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发表时间：
2021-12-01
期刊：
影响因子：
6.1
通讯作者：
Saber, Sameh
中科院分区：
文献类型：
--
作者：
Abdelhamid, Amir Mohamed;Youssef, Mahmoud E.;Saber, Sameh
目的：二甲双胍和恩格列净联合治疗可能具有互补作用，通过AMPK激活，超出其单药治疗的公认靶点。因此，本研究的目的是探讨首次在四氯化碳（CCl 4）诱导的小鼠肝纤维化模型，这种联合治疗的保肝作用。材料和方法：测定肝酶和肝脏的氧化应激参数，和羟脯氨酸含量进行生化。进行ELISA以测量PDGF-BB、TNF-α、TGF-β、TIMP 1、AMPK、p-mTOR、NF-κ B P65结合活性、p38 MAPK α、JNK 1/2和ERK 1/2。进行实时qPCR以测定Col 1a 1和a-SMA。此外，使用H&E和Masson三色染色进行组织病理学检查以确定组织病理学变化。关键发现：恩格列净抑制p38 MAPK和ERK 1/2的活化，并表现出弱AMPKa刺激。另一方面，二甲双胍对AMPKa产生更强的刺激作用，伴随NF-κ B核结合活性显著降低和p-mTOR水平下降。然而，二甲双胍对MAPK激酶的影响不显著。我们的研究结果表明，钝化p38 MAPKa和ERK 1/2活性恩格列净增强二甲双胍的抗纤维化作用，并增强其AMPK诱导的NF-κ B inactivation.Significance：由于糖尿病是肝纤维化最常见的危险因素之一，使用抗糖尿病药物有望改善治疗结果。因此，二甲双胍/恩格列净联合治疗通过显示出互补作用，特别是在糖尿病患者中，可能有希望预防肝脏炎症和纤维化。
Aim: Metformin and empagliflozin combined therapy may have complementary effects that go beyond the wellrecognized targets of their monotherapy through AMPK activation. Therefore, the current study was designed to investigate for the first time the hepatoprotective effects of such combination therapy in the carbon tetrachloride (CCl4)-induced hepatic fibrosis model in mice.Materials and methods: Determination of liver enzymes and the liver content of oxidative stress parameters, and hydroxyproline were performed biochemically. ELISA was performed to measure PDGF-BB, TNF-alpha, TGF-beta, TIMP1, AMPK, p-mTOR, NF-kappa B P65 binding activity, p38 MAPKa, JNK1/2 and ERK1/2. Real-time qPCR was conducted to determine Col1a1 and a-SMA. In addition, histopathological examination using H&E and Masson's trichrome stain were performed for determination of histopathological changes.Key findings: Empagliflozin inhibited the activation of p38 MAPK and ERK1/2 and exhibited a weak AMPKa stimulation. On the other hand, metformin exerted a more robust stimulatory action on the AMPKa that was accompanied by a notable decrease in the NF-kappa B nuclear binding activity and a decline in the p-mTOR levels. Nevertheless, the effect of metformin on MAPK kinases was insignificant. Our results revealed that blunting p38 MAPKa and ERK1/2 activities by empagliflozin enhanced the antifibrotic effect of metformin and augmented its AMPK-induced NF-kappa B inactivation.Significance: As diabetes is one of the most common risk factors for liver fibrosis, the use of antidiabetic drugs is expected to improve therapeutic outcome. Therefore, metformin/empagliflozin combined therapy could be promising in preventing hepatic inflammation and fibrosis via exhibiting complementary effects particularly in diabetic patients.