El Tabaa Maram Mohammed, Faheem Heba, Elballal Mohammed Salah, Rashad Eman, Mohsen Mohamed, El Tabaa Manar Mohammed
Physiology Department, Faculty of Medicine, Tanta University, Egypt.
Biochemistry, Faculty of Pharmacy, Badr University in Cairo (BUC), Badr City, Cairo, 11829, Egypt.
Eur J Pharmacol. 2025 Mar 15;991:177306. doi: 10.1016/j.ejphar.2025.177306. Epub 2025 Jan 27.
Liver damage is one of the most severe side effects of valproic acid (VPA) therapy. Research indicates that PPAR-α prevents Wnt3a/β-catenin-induced PGC-1α dysregulation, which is linked to liver injury. Although PPAR-α activation has hepatoprotective effects, its role in preventing VPA-induced liver injury remains unclear. Our research used network analysis, molecular docking, and in-vivo validation to predict and assess targets and pathways associated with the hepatoprotective effects of oleoylethanolamide (OEA), a PPAR-α agonist, on VPA-induced steatohepatitis. For in-vivo experiments, 24 rats were assigned to V, OEA, VPA, and OEA + VPA. Liver functions, TGs, cholesterol, and LDL were tested. Hepatic levels of PPAR-α, ACO, TNF-α, IL-1β, HO-1, MDA, and TAC, along with Wnt3a/β-catenin, PGC-1α, and Nrf2 expression were assessed. Further, NF-κB, Bax, Bcl-2, and caspase-3 expression were detected immunohistochemically. Network pharmacology identified 258 targets for OEA-steatohepatitis connection, including NFKB1, PPARA, and NFE2L2, in addition to TNF, non-alcoholic fatty liver, NF-κB, PPAR, and WNT signaling, as contributing to steatohepatitis pathogenesis. The docking revealed a strong affinity between OEA and Wnt3a, β-catenin, and PGC-1α. Therefore, we postulated that the hepatoprotective effect of OEA may be due to Wnt3a/β-catenin-mediated inactivation of PGC1-α pathway. In vivo, OEA inhibited Wnt3a/β-catenin and increased PGC1-α by activating PPAR-α. Hence, PGC1-α reduced fat cell β-oxidation and NF-κB-mediated inflammation. OEA lessened MDA and raised TAC to mitigate oxidative damage. OEA additionally reduced apoptosis by lowering Bax/Bcl-2 ratio and caspase-3. In summary, PPAR-α involvement in the protective effects of OEA against VPA-induced steatohepatitis can be confirmed by suppressing Wnt3a/β-catenin and activating PGC-1α signaling.
肝损伤是丙戊酸（VPA）治疗最严重的副作用之一。研究表明，过氧化物酶体增殖物激活受体α（PPAR-α）可防止Wnt3a/β-连环蛋白诱导的过氧化物酶体增殖物激活受体γ辅助激活因子1α（PGC-1α）失调，而这种失调与肝损伤有关。虽然PPAR-α激活具有肝脏保护作用，但其在预防VPA诱导的肝损伤中的作用仍不清楚。我们的研究使用网络分析、分子对接和体内验证来预测和评估油酰乙醇胺（OEA，一种PPAR-α激动剂）对VPA诱导的脂肪性肝炎的肝脏保护作用相关的靶点和通路。对于体内实验，将24只大鼠分为V组、OEA组、VPA组和OEA + VPA组。检测肝功能、甘油三酯（TGs）、胆固醇和低密度脂蛋白（LDL）。评估肝脏中PPAR-α、酰基辅酶A氧化酶（ACO）、肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、血红素加氧酶-1（HO-1）、丙二醛（MDA）和总抗氧化能力（TAC）水平，以及Wnt3a/β-连环蛋白、PGC-1α和核因子E2相关因子2（Nrf2）的表达。此外，通过免疫组织化学检测核因子κB（NF-κB）、 Bax、Bcl-2和半胱天冬酶-3（caspase-3）的表达。网络药理学确定了OEA与脂肪性肝炎关联的258个靶点，包括NFKB1、PPARA和NFE2L2，此外还有TNF、非酒精性脂肪肝、NF-κB、PPAR和WNT信号通路，这些都与脂肪性肝炎的发病机制有关。分子对接显示OEA与Wnt3a、β-连环蛋白和PGC-1α之间具有很强的亲和力。因此，我们推测OEA的肝脏保护作用可能是由于Wnt3a/β-连环蛋白介导的PGC1-α通路失活。在体内，OEA通过激活PPAR-α抑制Wnt3a/β-连环蛋白并增加PGC1-α。因此，PGC1-α减少脂肪细胞β-氧化和NF-κB介导的炎症。OEA降低MDA并提高TAC以减轻氧化损伤。OEA还通过降低Bax/Bcl-2比值和caspase-3减少细胞凋亡。总之，通过抑制Wnt3a/β-连环蛋白和激活PGC-1α信号通路，可以证实PPAR-α参与了OEA对VPA诱导的脂肪性肝炎的保护作用。
