Recombinant bactericidal/permeability-increasing protein attenuates the systemic inflammatory response syndrome in lower limb ischemia-reperfusion injury
Recombinant bactericidal/permeability-increasing protein attenuates the systemic inflammatory response syndrome in lower limb ischemia-reperfusion injury
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DOI：
10.1067/mva.2001.111992
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发表时间：
2001-04-01
影响因子：
4.3
通讯作者：
Campbell, FC
中科院分区：
文献类型：
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作者：
Harkin, DW;Barros, AAB;Campbell, FC
目的：后肢缺血再灌注(I/R)损伤增加肠道通透性，由此引起的内毒素血症与放大的全身炎症反应综合征相关，导致多器官功能障碍综合征。目的：研究重组杀菌/通透性增加蛋白(rBPI(21))在后肢I/R损伤后内毒素增强型全身炎症反应综合征中的作用。方法：48只体重300~350g的雄性Wistar大鼠随机分为假手术组(Sham)和双侧后肢缺血3h再灌流(I/R)2 h组(n=8)。对照组和未处理的I/R组给予2 mg/kg的对照蛋白制剂thaumatin。治疗组在再灌流开始时按1、2或4 mg/kg体重静脉注射rBPI(21)，另一组在再灌流1h后按2 mg/kg静脉注射rBPI(21)。用生物测定法测定血浆白介素6浓度作为全身炎症反应的指标。血浆内毒素浓度用无核细胞裂解物显色法测定。用酶联免疫吸附试验检测内毒素高度保守的核心区域的交叉反应性免疫球蛋白G和M抗体。结果：I/R组血浆IL-6浓度(1351.20 pg/m L[860.16-1886.40 pg/m L])显著高于对照组(125.32 p g/m L[87.76-157.52 pg/m L；P<；P<0.01]。再灌流时给予rBPI(21)2 mg/kg(715.89 pg/m L[573.36-847.76 pg/m L])，与非治疗组比较，IL-6反应显著降低(P<；.016)。缺血再灌注后血浆内毒素水平显著升高(21.52pg/mL[6.20~48.23pg/mL[P<；0.0001]，与对照组(0.90pg/mL[0.00~2.30pg/mL[P<；0.0001]相比；rBPI(21)4 mg/kg治疗组再灌流时血浆内毒素血症显著降低(1.30pg/mL[1.20~2.20pg/mL])，与未治疗组比较差异有统计学意义(P<；0.0001)。肺组织髓过氧化物酶活性：未治疗I/R组(208.18%[128.79%~221.81%])显著高于对照组(62.00%[40.45%~80.92%；P<；0.0001]；rBPI(21)2 mg/kg组(129.54%[90.49%~145.78%；P<；0.05])。结论：后肢缺血再灌注损伤与内毒素血症、血浆白介素6升高和肺白细胞计数有关。缺血后用rBPI(21)治疗可降低内毒素血症、白介素6反应，并减轻对后肢再灌流损伤所致的肺白细胞减少。
Objectives: Hind limb ischemia-reperfusion (I/R) injury increases gut permeability, and resultant endotoxemia is associated with an amplified systemic inflammatory response syndrome leading to multiple organ dysfunction syndrome. We studied the potential role of recombinant bactericidal/permeability-increasing protein (rBPI(21)), a novel antiendotoxin therapy, in modulating endotoxin-enhanced systemic inflammatory response syndrome in hind limb I/R injury.Methods: In this prospective, randomized, controlled, experimental animal study, 48 male Wistar rats, weighing 300 to 350 g, were randomized to a control group (sham) and five groups undergoing 3 hours bilateral hind limb ischemia with 2 hours reperfusion (I/R) (n = 8 per group). The control and untreated I/R groups received thaumatin, a control-protein preparation, at 2 mg/kg. Treatment groups were administered rBPI(21) intravenously at 1, 2, or 4 mg/kg body weight at the beginning of reperfusion; an additional group was administered rBPI(21) intravenously at 2 mg/kg after 1 hour of reperfusion. Plasma interleukin-6 concentration was estimated by bioassay as a measure of systemic inflammation. Plasma endotoxin concentration was determined by use of an amebocyte lysate chromogenic assay. Cross-reactive immunoglobulin G and M antibodies to the highly conserved inner core region of endotoxin were measured by use of an enzyme-linked immunosorbent assay. The lung tissue wet-to-dry weight ratio and myeloperoxidase concentration were used as markers of edema and neutrophil sequestration, respectively.Results: I/R provoked highly significant elevation in plasma interleukin-6 concentrations (1351.20 pg/mL [860.16 - 1886.40 pg/mL]) compared with controls (125.32 pg/mL [87.76-157.52 pg/mL; P < .0001]), but treatment with rBPI(21) 2 mg/kg at onset of reperfusion (715.89 pg/mL [573.36-847.76 pg/mL]) significantly decreased interleukin-6 response compared with the nontreatment group (P < .016). I/R increased plasma endotoxin concentrations significantly (21.52 pg/mL [6.20-48.23 pg/mL]), compared with control animals (0.90 pg/mL [0.00-2.30 pg/mL; P < .0001]), and treatment with rBPI(21) 4 mg/kg at reperfusion significantly decreased endotoxemia (1.30 pg/mL [1.20-2.20 pg/mL]), compared with the untreated group (P < .001). The lung tissue myeloperoxidase level was significantly increased in the untreated I/R group (208.18% [128.79%-221.81%]), compared with in controls (62.00% [40.45%-80.92%; P < .0001]), and attenuated in those treated with rBPI(21) 2 mg/kg (129.54% [90.49%-145.78%; P < .05]). Data represent median and interquartile range, comparisons made with the nonparametric Mann-Whitney U test.Conclusions: These findings show that hind limb ischemia-reperfusion injury is associated with endotoxemia, elevations in plasma interleukin-6, and pulmonary leukosequestration. Treatment with rBPI(21) after ischemia reduces endotoxemia, the interleukin-6 response, and attenuates pulmonary leukosequestration in response to hind limb reperfusion injury.