Sheppard Forest R, Moore Ernest E, Johnson Jeffrey L, Cheng Aaron M, McLaughlin Nathan, Silliman Christopher C
Department of Surgery, University of Colorado Health Sciences Center, Denver, Colorado 80204, USA.
J Trauma. 2004 Oct;57(4):720-4; discussion 724-5. doi: 10.1097/01.ta.0000140479.65350.28.
Red blood cell (pRBC) transfusion is an independent risk factor for multiple organ failure (MOF); a maladaptive immuno-inflammatory response is implicated. Interleukin-8 (IL-8) is one putative mediator of this response. We previously observed that injured patients resuscitated with pRBCs have increased plasma IL-8 compared with those given human polymerized hemoglobin (PolyHb). To further elucidate the mechanisms responsible for this difference in IL-8, we devised an ex-vivo transfusion model. We hypothesize that pRBC transfusion induces increased IL-8 gene expression that is avoided by the use of PolyHb.
Human volunteer blood was incubated alone (RB) or with a major transfusion (50% exchange) of either post-storage leukoreduced O-pRBCs (RB + pRBC) or PolyHb (RB + PolyHb) for 30 minutes at 37 degrees C. Total leukocyte (TL) or polymorphonuclear leukocyte (PMN) total RNA was isolated and IL-8 mRNA quantified. Results are reported as amol IL-8 mRNA/microg total RNA +/- SEM. Stats: ANOVA with Bonferroni/Dunn post hoc analysis.
Simulated transfusion of pRBCs increased TL IL-8 mRNA (RB=0.28 +/- 0.10 amol/microg total RNA, RB + pRBC=2.24 +/- 0.25 amol/microg total RNA, p <0.01), whereas PolyHb did not (B + PolyHb=0.82 +/- 0.30 amol/microg total RNA). PolyHb IL-8 mRNA was less than pRBC transfused (p <0.01). In PMNs, simulated transfusion of pRBCs increase IL-8 mRNA (RB=3.17 +/- 1.05 amol/microg total RNA, RB + pRBC=7.60 +/- 1.79 amol/microg total RNA, p <0.01), whereas PolyHb did not (RB + PolyHb=4.53 +/- 1.64 amol/microg total RNA).
Stored pRBCs induces increased TL and PMN IL-8 gene expression, whereas human polymerized hemoglobin, in lieu or pRBCs, avoids this increase. These experimental results corroborate our previous clinical studies and further encourage the study of PolyHb as a resuscitation strategy to decrease postinjury MOF.
红细胞（pRBC）输血是多器官功能衰竭（MOF）的一个独立危险因素；这与适应性免疫炎症反应失调有关。白细胞介素-8（IL-8）是这种反应的一种假定介质。我们之前观察到，与输注人聚合血红蛋白（PolyHb）的患者相比，接受pRBC复苏的受伤患者血浆IL-8水平升高。为了进一步阐明导致IL-8这种差异的机制，我们设计了一种体外输血模型。我们假设pRBC输血会诱导IL-8基因表达增加，而使用PolyHb可避免这种情况。
将人类志愿者血液单独孵育（RB），或在37℃下与主要输血（50%置换）的储存后白细胞滤除的O型pRBC（RB + pRBC）或PolyHb（RB + PolyHb）一起孵育30分钟。分离总白细胞（TL）或多形核白细胞（PMN）的总RNA并对IL-8 mRNA进行定量。结果以每微克总RNA中IL-8 mRNA的阿托摩尔数（amol）±标准误（SEM）报告。统计学分析：采用方差分析及Bonferroni/Dunn事后分析。
模拟输注pRBC可增加TL的IL-8 mRNA（RB = 0.28±0.10 amol/μg总RNA，RB + pRBC = 2.24±0.25 amol/μg总RNA，p<0.01），而PolyHb则不会（RB + PolyHb = 0.82±0.30 amol/μg总RNA）。PolyHb的IL-8 mRNA低于输注pRBC的情况（p<0.01）。在PMN中，模拟输注pRBC可增加IL-8 mRNA（RB = 3.17±1.05 amol/μg总RNA，RB + pRBC = 7.60±1.79 amol/μg总RNA，p<0.01），而PolyHb则不会（RB + PolyHb = 4.53±1.6 amol/μg总RNA）。
储存的pRBC可诱导TL和PMN的IL-8基因表达增加，而人聚合血红蛋白替代pRBC则可避免这种增加。这些实验结果证实了我们之前的临床研究，并进一步鼓励对PolyHb作为一种复苏策略进行研究，以降低损伤后MOF的发生率。
