Unit of Cellular and Molecular Pharmacology and Toxicology, Department of Internal Medicine and Therapeutics, University of Pavia, 27100 Pavia, Italy.
Department of Drug Science and Technology, University of Turin, 10125 Turin, Italy.
Int J Mol Sci. 2021 Feb 24;22(5):2234. doi: 10.3390/ijms22052234.
We previously demonstrated that the blockade of mGluR5 by 2-methyl-6(phenylethynyl)pyridine (MPEP) reduces both cold and warm ischemia/reperfusion injury. Here we evaluated whether MPEP reduces the hepatic preservation injury in rat livers from cardiac-death-donors (DCDs). Livers from DCD rats were isolated after an in situ warm ischemia (30 min) and preserved for 22 h at 4 °C with UW solution. Next, 10 mg/Kg MPEP or vehicle were administered 30 min before the portal clamping and added to the UW solution (3 µM). LDH released during washout was quantified. Liver samples were collected for iNOS, eNOS, NO, TNF-α, ICAM-1, caspase-3 and caspase-9 protein expression and nuclear factor-erythroid-2-related factor-2 (Nrf2) gene analysis. Lower LDH levels were detected in control grafts versus DCD groups. An increase in eNOS and NO content occurred after MPEP treatment; iNOS and TNF-α content was unchanged. ICAM-1 expression was reduced in the MPEP-treated livers as well as the levels of caspase-3 and caspase-9. Nrf2, oxidative stress-sensitive gene, was recovered to control value by MPEP. These results suggest that MPEP can be used to reclaim DCD livers subjected to an additional period of cold ischemia during hypothermic storage. MPEP protects against apoptosis and increased eNOS, whose overexpression has been previously demonstrated to be protective in hepatic ischemia/reperfusion damage.
我们之前的研究表明,mGluR5 阻断剂 2-甲基-6-(苯乙炔基)吡啶(MPEP)可减轻冷缺血再灌注损伤和热缺血再灌注损伤。在此,我们评估了 MPEP 是否可以减轻心脏死亡供体(DCD)大鼠肝脏的保存损伤。在原位热缺血(30 分钟)后,从 DCD 大鼠中分离出肝脏,并在 4°C 下用 UW 溶液保存 22 小时。然后,在门静脉夹闭前 30 分钟给予 10 mg/Kg MPEP 或载体,并将其添加到 UW 溶液(3 µM)中。洗脱过程中释放的 LDH 进行定量。收集肝组织样本,用于检测 iNOS、eNOS、NO、TNF-α、ICAM-1、caspase-3 和 caspase-9 蛋白表达和核因子-红细胞 2 相关因子-2(Nrf2)基因分析。与 DCD 组相比,对照组移植物中的 LDH 水平较低。MPEP 处理后 eNOS 和 NO 含量增加;iNOS 和 TNF-α 含量不变。MPEP 处理的肝脏中 ICAM-1 表达减少,caspase-3 和 caspase-9 的水平也降低。Nrf2,一种对氧化应激敏感的基因,通过 MPEP 恢复到对照值。这些结果表明,MPEP 可用于回收在低温保存期间经历额外冷缺血期的 DCD 肝脏。MPEP 可防止细胞凋亡和 eNOS 过度表达,先前的研究表明,eNOS 过度表达可保护肝脏缺血再灌注损伤。