Lehnert Mark, Uehara Tetsuya, Bradford Blair U, Lind Henrik, Zhong Zhi, Brenner David A, Marzi Ingo, Lemasters John J
Medical Univ. of South Carolina, 280 Calhoun St., PO Box 250140, Charleston, SC 29425, USA.
Am J Physiol Gastrointest Liver Physiol. 2006 Sep;291(3):G456-63. doi: 10.1152/ajpgi.00480.2005. Epub 2006 Apr 13.
Hemorrhagic shock and resuscitation cause endotoxemia and hepatocellular damage. Because lipopolysaccharide-binding protein (LBP) enhances cellular responses to endotoxin, our aim was to determine whether LBP contributes to hemorrhage/resuscitation-induced injury by comparing LBP knockout and wild-type mice. Under pentobarbital anaesthesia, wild-type and LBP-deficient mice were hemorrhaged to 30 mmHg for 3 h and then resuscitated with shed blood plus half the volume of lactated Ringer solution. Serum alanine aminotransferase (ALT) necrosis, neutrophil infiltration, and 4-hydroxynonenal by histology/cytochemistry and stress kinase activation by immunoblot analysis were then determined. ALT in wild-type mice was 2,461 +/- 383 and 1,418 +/- 194 IU/l (means +/- SE), respectively, at 2 and 6 h after resuscitation versus sham ALT of 102 +/- 6 IU/l. In LBP-deficient mice, ALT was blunted at both time points to 1,108 +/- 340 and 619 +/- 171 IU/l (P < 0.05). Liver necrosis after 6 h was also attenuated from 3.5 +/- 0.8% in wild-type mice to 1.3 +/- 0.5% in LBP-deficient mice (P < 0.05). After hemorrhage/resuscitation, neutrophil infiltration increased 71% more in wild-type than LBP knockout mice. Similarly, hepatic 4-hydroxynonenal staining, indicative of lipid peroxidation, decreased from 33.8 +/- 4.5% in wild-type mice to 11.6 +/- 1.9% in knockout mice (P < 0.05). After hemorrhage/resuscitation, activation of MAPKs, JNK and ERK, occurred in wild-type mice, which was largely blocked in LBP-deficient mice. However, endotoxin in portal blood after resuscitation was not significantly different between wild-type and knockout mice. In conclusion, hemorrhagic shock and resuscitation to mice cause severe, LBP-mediated hepatocellular damage. An absence of LBP blunts hepatocellular injury with decreased neutrophil infiltration, oxidative stress, and c-Jun and ERK activation.
失血性休克及复苏可导致内毒素血症和肝细胞损伤。由于脂多糖结合蛋白(LBP)可增强细胞对内毒素的反应,我们的目的是通过比较LBP基因敲除小鼠和野生型小鼠,来确定LBP是否参与出血/复苏诱导的损伤。在戊巴比妥麻醉下,将野生型和LBP缺陷型小鼠出血至血压30 mmHg并持续3小时,然后用自体失血加一半体积的乳酸林格液进行复苏。随后通过组织学/细胞化学测定血清丙氨酸转氨酶(ALT)、坏死情况、中性粒细胞浸润以及4-羟基壬烯醛,并通过免疫印迹分析测定应激激酶激活情况。复苏后2小时和6小时,野生型小鼠的ALT分别为2461±383和1418±194 IU/l(均值±标准误),而假手术组的ALT为102±6 IU/l。在LBP缺陷型小鼠中,两个时间点的ALT均降低至1108±340和619±171 IU/l(P<0.05)。6小时后的肝坏死也从野生型小鼠的3.5±0.8%减轻至LBP缺陷型小鼠的1.3±0.5%(P<0.05)。出血/复苏后,野生型小鼠中性粒细胞浸润比LBP基因敲除小鼠多增加71%。同样,指示脂质过氧化的肝脏4-羟基壬烯醛染色从野生型小鼠的33.8±4.5%降至基因敲除小鼠的11.6±1.9%(P<0.05)。出血/复苏后,野生型小鼠中丝裂原活化蛋白激酶(MAPKs)、应激活化蛋白激酶(JNK)和细胞外信号调节激酶(ERK)被激活,而在LBP缺陷型小鼠中这种激活在很大程度上被阻断。然而,复苏后野生型小鼠和基因敲除小鼠门静脉血中的内毒素没有显著差异。总之,对小鼠进行失血性休克及复苏会导致严重的、由LBP介导的肝细胞损伤。缺乏LBP可减轻肝细胞损伤,同时中性粒细胞浸润、氧化应激以及c-Jun和ERK激活减少。