Matrix Metalloproteinase-2 (MMP-2) Gene Deletion Enhances MMP-9 Activity, Impairs PARP-1 Degradation, and Exacerbates Hepatic Ischemia and Reperfusion Injury in Mice.
Matrix Metalloproteinase-2 (MMP-2) Gene Deletion Enhances MMP-9 Activity, Impairs PARP-1 Degradation, and Exacerbates Hepatic Ischemia and Reperfusion Injury in Mice.
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DOI：
10.1371/journal.pone.0137642
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Coito AJ
中科院分区：
文献类型：
--
作者：
Kato H;Duarte S;Liu D;Busuttil RW;Coito AJ
肝脏缺血再灌注损伤(IRI)是一种炎症状态，也是术后发病率和死亡率的重要原因。基质金属蛋白酶(MMPs)广泛参与炎症性疾病的发病机制。在不同的MMPs中，明胶酶(MMP2和MMP9)是肝脏IRI中最显著的MMPs。虽然基质金属蛋白酶-9在肝脏损伤中的作用已有相当多的文献记载，但有关基质金属蛋白酶-2活性在肝脏缺血再灌注损伤中的作用的直接证据仍有待建立。由于在体内缺乏合适的针对单个MMPs的抑制剂，基因操作是评估MMPs对肝损伤的直接贡献的必要工具。因此，我们使用了MMP2-/-缺陷小鼠和MMP2+/+野生型仔鼠来研究MMP2活性在肝脏IRI中的作用。在手术前后的野生型肝脏的肝窦和IRI后的少量白细胞中检测到了基质金属蛋白酶-2的表达。与MMP2+/+小鼠相比，MMP2缺失(MMP2-/-)小鼠在再灌注后6、24和48小时表现出加重的肝损伤，在某些情况下是致命的。在体外和肝脏IRI后，MMP2缺乏导致MMP9活性上调，自发性白细胞在幼稚的肝脏中渗透，并增加依赖于MMP9的白细胞的迁移。此外，基质金属蛋白酶-2活性的完全丧失损害了再灌注后严重受损的肝脏中聚(ADP-核糖)聚合酶(PARP-1)的降解。然而，给予MMP2缺失的小鼠注射PARP-1抑制剂后，肝脏保存恢复到几乎与IRI后MMP2+/+小鼠相当的水平。通过测量培养液中乳酸脱氢酶的外流来评估基质金属蛋白酶-2缺失的肝窦内皮细胞的PARP-1降解缺陷与其增加的细胞毒性相关。综上所述，我们的研究结果首次表明，基质金属蛋白酶-2基因缺失加剧了肝脏IRI。此外，他们对基质金属蛋白酶-2对炎症反应的调控提供了新的见解，这可能与设计新的以基质金属蛋白酶为靶向的药物来治疗肝脏IRI有关。
Hepatic ischemia and reperfusion injury (IRI) is an inflammatory condition and a significant cause of morbidity and mortality after surgery. Matrix metalloproteinases (MMPs) have been widely implicated in the pathogenesis of inflammatory diseases. Among the different MMPs, gelatinases (MMP-2 and MMP-9) are within the most prominent MMPs detected during liver IRI. While the role of MMP-9 in liver damage has been fairly documented, direct evidence of the role for MMP-2 activity in hepatic IRI remains to be established. Due to the lack of suitable inhibitors to target individual MMPs in vivo, gene manipulation is as an essential tool to assess MMP direct contribution to liver injury. Hence, we used MMP-2-/- deficient mice and MMP-2+/+ wild-type littermates to examine the function of MMP-2 activity in hepatic IRI. MMP-2 expression was detected along the sinusoids of wild-type livers before and after surgery and in a small population of leukocytes post-IRI. Compared to MMP-2+/+ mice, MMP-2 null (MMP-2-/-) mice showed exacerbated liver damage at 6, 24, and 48 hours post-reperfusion, which was fatal in some cases. MMP-2 deficiency resulted in upregulation of MMP-9 activity, spontaneous leukocyte infiltration in naïve livers, and amplified MMP-9-dependent transmigration of leukocytes in vitro and after hepatic IRI. Moreover, complete loss of MMP-2 activity impaired the degradation of poly (ADP-ribose) polymerase (PARP-1) in extensively damaged livers post-reperfusion. However, the administration of a PARP-1 inhibitor to MMP-2 null mice restored liver preservation to almost comparable levels of MMP-2+/+ mice post-IRI. Deficient PARP-1 degradation in MMP-2-null sinusoidal endothelial cells correlated with their increased cytotoxicity, evaluated by the measurement of LDH efflux in the medium. In conclusion, our results show for the first time that MMP-2 gene deletion exacerbates liver IRI. Moreover, they offer new insights into the MMP-2 modulation of inflammatory responses, which could be relevant for the design of new pharmacological MMP-targeted agents to treat hepatic IRI.