Li Bin, Di Guohu, Ge Huanhuan, Song Peirong, Han Wenshuo, Sun Hetong, Wang Dianqiang, Chen Peng, Wang Ye
School of Basic Medicine, Qingdao University, Qingdao, Shandong Province 266071, China.
School of Basic Medicine, Qingdao University, Qingdao, Shandong Province 266071, China; Institute of Stem Cell Regeneration Medicine, School of Basic Medicine, Qingdao University, Qingdao, Shandong Province 266071, China.
Cell Signal. 2025 Mar;127:111602. doi: 10.1016/j.cellsig.2025.111602. Epub 2025 Jan 13.
During the proliferative phase of liver regeneration, insufficient regulation of hepatocyte hydrogen peroxide (HO) overproduction can result in oxidative stress and hepatocyte death. This study aims to investigate the influence of Aquaporin 5 (Aqp5) on liver regeneration by evaluating its role in reactive oxygen species (ROS) generation and NLRP3-GSDMD-mediated pyroptosis. A 70 % partial hepatectomy (PHx) model was established in Aqp5 mice to evaluate the pathological changes in the liver. Reactive oxygen species (ROS) production was assessed using a dichlorodihydrofluorescein diacetate (DCFH-DA) assay. Aqp5 deficiency significantly increased ROS production, the number of TUNEL-positive cells, and disrupted mitochondrial membrane potential in the liver of Aqp5-deficient mice. The impact of Aqp5 on ROS/NLRP3/Gasdermin-D (GSDMD)-mediated pyroptosis was examined through the administration of N-acetyl-L-cysteine (NAC, an ROS scavenger) or disulfiram (DSF, a GSDMD inhibitor). In Aqp5-deficient mice, the regenerative liver exhibited increased expression of NLRP3, enhanced activation of caspase-1 and GSDMD, as well as elevated secretion of IL-1β. Treatment with DSF significantly attenuated GSDMD-mediated pyroptosis triggered by Aqp5 deficiency in the regenerating liver. Furthermore, the administration of NAC to Aqp5-deficient mice resulted in a reduction in the expression levels of NLRP3, the activity levels of caspase-1 and GSDMD, as well as the release of IL-1β. Our findings indicate that the deficiency of Aqp5 facilitates GSDMD activation through the production of ROS. The suppression of ROS or inhibition of GSDMD significantly alleviates the damage and pyroptosis observed in Aqp5-deficient regenerative liver.
在肝脏再生的增殖阶段，肝细胞过氧化氢（HO）产生过多而调节不足会导致氧化应激和肝细胞死亡。本研究旨在通过评估水通道蛋白5（Aqp5）在活性氧（ROS）生成以及NLRP3 - GSDMD介导的细胞焦亡中的作用，来研究其对肝脏再生的影响。在Aqp5基因敲除小鼠中建立70%部分肝切除术（PHx）模型，以评估肝脏的病理变化。使用二氯二氢荧光素二乙酸酯（DCFH - DA）检测法评估活性氧（ROS）的产生。Aqp5基因缺失显著增加了Aqp5基因敲除小鼠肝脏中的ROS产生、TUNEL阳性细胞数量，并破坏了线粒体膜电位。通过给予N - 乙酰 - L - 半胱氨酸（NAC，一种ROS清除剂）或双硫仑（DSF，一种GSDMD抑制剂），研究了Aqp5对ROS/NLRP3/ Gasdermin - D（GSDMD）介导的细胞焦亡的影响。在Aqp5基因敲除小鼠中，再生肝脏中NLRP3的表达增加，半胱天冬酶 - 1和GSDMD的激活增强，以及IL - 1β的分泌增加。用DSF治疗可显著减轻再生肝脏中由Aqp5基因缺失引发的GSDMD介导的细胞焦亡。此外，给Aqp5基因敲除小鼠注射NAC导致NLRP3的表达水平、半胱天冬酶 - 1和GSDMD的活性水平以及IL - 1β的释放降低。我们的研究结果表明，Aqp5的缺乏通过ROS的产生促进了GSDMD的激活。抑制ROS或抑制GSDMD可显著减轻Aqp5基因敲除的再生肝脏中观察到的损伤和细胞焦亡。
