US Army Institute of Surgical Research, JBSA Fort Sam Houston, San Antonio, Texas.
Shock. 2017 Oct;48(4):449-458. doi: 10.1097/SHK.0000000000000865.
The use of albumin for resuscitation has not proven as beneficial in human trials as expected from numerous animal studies. One explanation could be the practice of adding fatty acid (FA) during manufacture of pharmaceutical albumin. During ischemia, unbound free FAs (FFA) in the circulation could potentially induce cellular damage. We hypothesized that albumins with higher available binding capacities (ABC) for FFAs may prevent that damage. Therefore, we developed a technique to measure ABC, determined if pharmaceutical human serum albumin (HSA) has decreased ABC compared with FA-free bovine serum albumin (BSA), and if binding capacity would affect hemolysis when blood is mixed with exogenous FFA at levels similar to those observed in shock. The new assay used exogenous oleic acid (OA), glass fiber filtration, and a FFA assay kit. RBC hemolysis was determined by mixing 0 to 5 mM OA with PBS, HSA, FA-free BSA, or FA-saturated BSA and measuring plasma hemoglobin after incubation with human blood. 5% HSA contained 4.7±0.2 mM FFA, leaving an ABC of 5.0 ± 0.6 mM, compared with FA-free BSAs ABC of 7.0 ± 1.3 mM (P < 0.024). Hemolysis after OA was reduced with FA-free BSA but increased with FA-saturated BSA. HSA provided intermediate results. 25% solutions of FA-free BSA and HSA were more protective, while 25% FA-saturated BSA was more damaging than 5% solutions. These findings suggest that increased FA saturation may reverse albumin's potential benefit to lessen cellular damage and may explain, at least in part, its failure in human trauma studies.
在人体试验中,白蛋白的复苏效果并不像许多动物研究预期的那样有益。一种解释可能是在制药白蛋白生产过程中添加脂肪酸 (FA) 的做法。在缺血期间,循环中未结合的游离脂肪酸 (FFA) 可能潜在地诱导细胞损伤。我们假设具有更高游离脂肪酸结合能力 (ABC) 的白蛋白可能会防止这种损伤。因此,我们开发了一种测量 ABC 的技术,确定与无 FA 的牛血清白蛋白 (BSA) 相比,药物人血清白蛋白 (HSA) 的 ABC 是否降低,以及如果结合能力在血液与外源性 FFA 混合时会影响溶血,其水平类似于休克中观察到的水平。新的测定方法使用外源性油酸 (OA)、玻璃纤维过滤和游离脂肪酸测定试剂盒。通过将 0 至 5mM OA 与 PBS、HSA、无 FA 的 BSA 或 FA 饱和的 BSA 混合,并在与人血孵育后测量血浆血红蛋白来确定 RBC 溶血。5%的 HSA 含有 4.7±0.2mM FFA,ABC 为 5.0±0.6mM,而无 FA 的 BSA 的 ABC 为 7.0±1.3mM(P<0.024)。OA 后的溶血用无 FA 的 BSA 减少,但用 FA 饱和的 BSA 增加。HSA 提供了中间结果。无 FA 的 BSA 和 HSA 的 25%溶液更具保护作用,而 25%FA 饱和的 BSA 比 5%溶液更具破坏性。这些发现表明,增加 FA 饱和度可能会逆转白蛋白减轻细胞损伤的潜在益处,并且至少部分解释了其在人体创伤研究中失败的原因。