Beta-arrestin 2 negatively regulates sepsis-induced inflammation
Beta-arrestin 2 negatively regulates sepsis-induced inflammation
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DOI：
10.1111/j.1365-2567.2009.03185.x
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发表时间：
2010-07-01
期刊：
影响因子：
6.4
通讯作者：
Cook, James A.
中科院分区：
文献类型：
--
作者：
Fan, Hongkuan;Bitto, Alessandra;Cook, James A.
β-抑制蛋白1和2是广泛表达的蛋白质，通过G蛋白偶联受体改变信号传导。β-抑制蛋白2在调节细胞炎症反应中作为信号传导衔接子和支架发挥重要作用。我们假设β-arrestin 2是实验性脓毒症炎症反应的关键调节因子。对β-抑制蛋白2（-/-）和野生型（WT）小鼠进行盲肠结扎和穿孔（CLP）。与WT小鼠（53%存活率）相比，β-抑制蛋白2（-/-）小鼠（13%存活率）的存活率显著降低（P < 0，中心点05）。在CLP后18小时处死第二组小鼠，用于血液、腹膜灌洗和组织样品收集。与WT小鼠相比，β-抑制蛋白2（-/-）中CLP诱导的血浆白细胞介素（IL）-6显著增加25 +/- 12倍，盲肠髓过氧化物酶（MPO）活性增加2中心点4 +/- 0中心点3倍。通过组织病理学和菌落形成单位计数测量，与CLP攻击后的WT小鼠相比，β-抑制蛋白2（-/-）小鼠表现出更严重的肺损伤和更高的细菌负荷。在随后的实验中，从β-抑制蛋白2（-/-）和WT小鼠中分离和培养脾细胞、腹膜巨噬细胞和骨髓衍生的巨噬细胞（BMDM），并在体外用脂多糖（LPS）刺激。LPS诱导的肿瘤坏死因子（TNF-α）、IL-6和IL-10的产生显著增加（分别为2个中心点2 +/-0中心点2倍、1个中心点8 +/-0中心点1倍和2个中心点2 +/-0中心点4倍;与WT小鼠相比，β-抑制蛋白2（-/-）小鼠的脾细胞中P < 0（中心点05）。脾细胞反应与腹膜巨噬细胞或BMDM的反应不同，其在WT和β-抑制蛋白2（-/-）小鼠之间在LPS刺激后的TNF-α和IL-6产生方面没有差异。我们的数据表明，β-arrestin 2的功能，负调控炎症反应的多微生物败血症。
P>beta-arrestins 1 and 2 are ubiquitously expressed proteins that alter signalling by G-protein-coupled receptors. beta-arrestin 2 plays an important role as a signalling adaptor and scaffold in regulating cellular inflammatory responses. We hypothesized that beta-arrestin 2 is a critical modulator of inflammatory response in experimental sepsis. beta-arrestin 2(-/-) and wild-type (WT) mice were subjected to caecal ligation and puncture (CLP). The survival rate was significantly decreased (P < 0 center dot 05) in beta-arrestin 2(-/-) mice (13% survival) compared with WT mice (53% survival). A second group of mice were killed 18 hr after CLP for blood, peritoneal lavage and tissue sample collection. CLP-induced plasma interleukin (IL)-6 was significantly increased 25 +/- 12 fold and caecal myeloperoxidase (MPO) activity was increased 2 center dot 4 +/- 0 center dot 3 fold in beta-arrestin 2(-/-) compared with WT mice. beta-arrestin 2(-/-) mice exhibited more severe lung damage and higher bacterial loads compared with WT mice post CLP challenge as measured by histopathology and colony-forming unit count. In subsequent experiments, splenocytes, peritoneal macrophages and bone marrow-derived macrophages (BMDMs) were isolated and cultured from beta-arrestin 2(-/-) and WT mice and stimulated in vitro with lipopolysaccharide (LPS). Tumour necrosis factor (TNF)-alpha, IL-6 and IL-10 production induced by LPS was significantly augmented (2 center dot 2 +/- 0 center dot 2 fold, 1 center dot 8 +/- 0 center dot 1 fold, and 2 center dot 2 +/- 0 center dot 4 fold, respectively; P < 0 center dot 05) in splenocytes from beta-arrestin 2(-/-) mice compared with WT mice. The splenocyte response was different from that of peritoneal macrophages or BMDMs, which exhibited no difference in TNF-alpha and IL-6 production upon LPS stimulation between WT and beta-arrestin 2(-/-) mice. Our data demonstrate that beta-arrestin 2 functions to negatively regulate the inflammatory response in polymicrobial sepsis.