Lieberman J M, Sacchettini J, Marks C, Marks W H
Department of Organ Transplantation, Swedish Medical Center, Seattle, Wash. 98104, USA.
Surgery. 1997 Mar;121(3):335-42. doi: 10.1016/s0039-6060(97)90363-9.
Human intestinal fatty acid binding protein (hIFABP) is a cytoplasmic protein of mature small intestinal epithelium. Work with the rat demonstrated that serum levels of IFABP correlated with early phases of intestinal mucosal injury. The aim of this study was to develop an assay for hIFABP and assess its usefulness as a marker for intestinal mucosal injury in human beings.
Recombinant hIFABP (r-hIFABP) was used to produce rabbit anti-hIFABP. Specificity and avidity of binding were tested with immunoprecipitation and Scatchard analysis. r-hIFABP was labeled with 125I, and a competitive assay was developed. Urine and serum from normal volunteers and from patients with necrotizing enterocolitis (NEC), acute thromboembolic related intestinal ischemia, and systemic inflammatory response syndrome were tested for hIFABP.
Molecular weight was 10(-12) kd, limit of detection was 1.87 ng/ml, and no cross-reactivity occurred when tested against rat IFABP or human heart FABP. Mean levels of hIFABP (ng/ml) were controls (serum less than 1.87, urine less than 1.87), NEC (serum 14.7 ng/ml), intestinal ischemia (serum 50 ng/ml, urine 52.3 ng/ml), systemic inflammatory response syndrome (serum 5.3 ng/ml, urine 13.2 ng/ml).
This assay is quantitative for hIFABP in serum and urine. Results from both normal persons and those with various causes of intestinal ischemia parallel our previous findings in the rat. Preliminary findings suggest that hIFABP may serve as a diagnostic marker for early intestinal mucosal compromise and, in addition, that it should prove useful as a tool in developing rationale therapeutic regimens to treat these complex clinical problems.
人肠脂肪酸结合蛋白(hIFABP)是成熟小肠上皮细胞的一种胞质蛋白。对大鼠的研究表明,IFABP的血清水平与肠黏膜损伤的早期阶段相关。本研究的目的是开发一种检测hIFABP的方法,并评估其作为人类肠黏膜损伤标志物的实用性。
使用重组hIFABP(r-hIFABP)制备兔抗hIFABP。通过免疫沉淀和Scatchard分析检测结合的特异性和亲和力。用125I标记r-hIFABP,并开发了一种竞争性检测方法。对正常志愿者以及患有坏死性小肠结肠炎(NEC)、急性血栓栓塞相关肠缺血和全身炎症反应综合征患者的尿液和血清进行hIFABP检测。
分子量为10(-12)kd,检测限为1.87 ng/ml,与大鼠IFABP或人心脏FABP检测时无交叉反应。hIFABP的平均水平(ng/ml)为:对照组(血清<1.87,尿液<1.87),NEC(血清14.7 ng/ml),肠缺血(血清50 ng/ml,尿液52.3 ng/ml),全身炎症反应综合征(血清5.3 ng/ml,尿液13.2 ng/ml)。
该检测方法可对血清和尿液中的hIFABP进行定量。正常人和各种原因引起肠缺血患者的检测结果与我们之前在大鼠中的发现一致。初步研究结果表明,hIFABP可能作为早期肠黏膜损伤的诊断标志物,此外,它应被证明是制定合理治疗方案以治疗这些复杂临床问题的有用工具。