Zhang Lu, Wang Mingfu, An Ran, Dai Jun, Liu Shujun, Chen Ming, Ding Haoran
Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, Jiangsu, China.
Surgery Department I, Zhangjiagang Traditional Chinese Medicine Hospital, Suzhou, Jiangsu, China.
J Clin Transl Hepatol. 2023 Oct 28;11(5):1069-1078. doi: 10.14218/JCTH.2022.00109. Epub 2023 May 23.
Donors with fatty livers are considered to address the shortage of livers for transplantation, but those livers are particularly sensitive to ischemia-reperfusion injury (IRI), and an increased incidence of graft failure is observed. Kupffer cells account for 20-35% of liver nonparenchymal cells, and have been shown to participate in the process of IRI and inflammatory reactions of hepatic steatosis. NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) is an intracellular sensor activated by Kupffer cells to promote generation and participates in IRI. Dynamics-associated protein 1 (Drp1) is one of the main proteins regulating mitochondrial division and exacerbates IRI by affecting mitochondrial dynamics. The mechanism of interaction of Kupffer cells with Drp1 and NLRP3 to aggravate IRI has not been clarified.
A mouse model of hepatic steatosis was established by feeding the mice with a high-fat diet. experiments were performed using AML12 normal mouse liver cells and RAW264.7 mononuclear macrophage cells cultured in medium with palmitate and oleic acid. Western blotting and immunohistochemical (IHC) staining were used to detect the expression of NLRPP3 and Drp1 in IRI in the control and high-fat diet groups. The expression of F4/80+ cells during IRI in hepatic steatosis was verified by IHC staining, and the role of NLRPP3 and Drp1 in Kupffer-cell mediated IRI was investigated by targeting Drp-1 inhibition.
Drp1 and NLRP3 expression was increased during IRI in hepatic steatosis, and the expression of Drp1 and NLRP3 were decreased after the elimination of Kupffer cells. That indicated Kupffer cells were involved in the process of IRI in hepatic steatosis through the action of Drp1 and NLRP3. After Drp1 inhibition, liver function was restored and NLRP3 expression level was reduced.
Kupffer cells aggravated IRI in hepatic steatosis via NLRP3 and Drp1. Drp1 inhibitors might be useful as specific therapeutics to alleviate IRI in hepatic steatosis and may have promise in case of liver donor shortage.
脂肪肝供体被认为可解决肝移植供肝短缺问题，但这些肝脏对缺血再灌注损伤（IRI）特别敏感，且观察到移植失败发生率增加。库普弗细胞占肝脏非实质细胞的20%-35%，已证明其参与IRI过程及肝脂肪变性的炎症反应。NOD样受体热蛋白结构域相关蛋白3（NLRP3）是一种由库普弗细胞激活的细胞内传感器，可促进炎症小体的生成并参与IRI。动力相关蛋白1（Drp1）是调节线粒体分裂的主要蛋白之一，通过影响线粒体动力学加重IRI。库普弗细胞与Drp1和NLRP3相互作用加重IRI的机制尚未阐明。
通过给小鼠喂食高脂饮食建立肝脂肪变性小鼠模型。使用在含有棕榈酸和油酸的培养基中培养的AML12正常小鼠肝细胞和RAW264.7单核巨噬细胞进行实验。采用蛋白质免疫印迹法和免疫组织化学（IHC）染色检测对照组和高脂饮食组IRI中NLRPP3和Drp1的表达。通过IHC染色验证肝脂肪变性IRI期间F4/80+细胞的表达，并通过靶向Drp-1抑制研究NLRPP3和Drp1在库普弗细胞介导的IRI中的作用。
肝脂肪变性IRI期间Drp1和NLRP3表达增加，消除库普弗细胞后Drp1和NLRP3表达降低。这表明库普弗细胞通过Drp1和NLRP3的作用参与肝脂肪变性的IRI过程。抑制Drp1后，肝功能恢复，NLRP3表达水平降低。
库普弗细胞通过NLRP3和Drp1加重肝脂肪变性中的IRI。Drp1抑制剂可能作为缓解肝脂肪变性中IRI的特异性治疗药物，在肝供体短缺的情况下可能具有应用前景。
