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使用艾考糊精进行透析会干扰血清α淀粉酶活性的测定。

Dialysis with icodextrin interferes with measurement of serum alpha-amylase activity.

作者信息

Schoenicke Gerrit, Grabensee Bernd, Plum Joerg

机构信息

Department of Nephrology and Rheumatology, H. Heine University Duesseldorf, Germany.

出版信息

Nephrol Dial Transplant. 2002 Nov;17(11):1988-92. doi: 10.1093/ndt/17.11.1988.

Abstract

BACKGROUND

The glucose polymer icodextrin has gained a widespread use in peritoneal dialysis especially in patients with low ultrafiltration and high peritoneal transport properties. In patients using a once-daily exchange with icodextrin, a decreased serum amylase activity has been reported. We explored the potential underlying mechanisms of this effect.

METHODS

Using standard chromolytic methods, serum amylase activity was measured in blood samples from 11 patients on icodextrin treatment and from 11 patients on conventional glucose treatment. Samples were additionally supplemented with alpha-amylase and unused icodextrin dialysis fluid. Potential complex formation between icodextrin and alpha-amylase was studied by SDS-gel electrophoresis with protein silver staining and fluorophore-assisted carbohydrate staining with the AMAC (FACE) method, which provides oligosaccharide labelling. Lipase activity was measured in parallel in all samples by two standard methods.

RESULTS

Amylase activity was reduced by 90% in serum from patients using icodextrin for the long dwell (15.9+/-10.9 U/l) vs patients using standard glucose (157.1+/-23.7 U/l; P<0.001). Addition of icodextrin to serum samples from patients using conventional glucose solutions induced a dose-dependent decrease in amylase activity. The assay results indicated a substrate competition between ET7-G7PNP and icodextrin. AMAC fluorophore staining of icodextrin and subsequent gel electrophoresis failed to demonstrate complex formation between icodextrin and alpha-amylase. Unlike the amylase findings, icodextrin did not affect lipase activity.

CONCLUSIONS

The present findings indicate that icodextrin competitively interacts as a substrate in the amylase assay. In support of this, fluorophore-assisted oligosaccharide electrophoresis on SDS gel failed to reveal the formation of an 'amylase/icodextrin complex'. Lipase measurement should provide an alternative and unconfounded method for diagnosing pancreatitis in icodextrin patients.

摘要

背景

葡萄糖聚合物异麦芽糖糊精在腹膜透析中已得到广泛应用,尤其适用于超滤量低且腹膜转运特性高的患者。据报道,在使用异麦芽糖糊精每日一次交换治疗的患者中,血清淀粉酶活性降低。我们探讨了这种效应的潜在机制。

方法

采用标准比色法,对11例接受异麦芽糖糊精治疗的患者和11例接受传统葡萄糖治疗的患者的血样进行血清淀粉酶活性检测。样本还补充了α -淀粉酶和未使用的异麦芽糖糊精透析液。通过十二烷基硫酸钠 - 凝胶电泳(SDS - gel electrophoresis)结合蛋白质银染以及使用AMAC(荧光辅助碳水化合物染色法,FACE)进行寡糖标记的荧光团辅助碳水化合物染色法,研究异麦芽糖糊精与α -淀粉酶之间潜在的复合物形成。通过两种标准方法同时测定所有样本中的脂肪酶活性。

结果

长期使用异麦芽糖糊精的患者血清中淀粉酶活性降低了90%(15.9±10.9 U/l),而使用标准葡萄糖的患者血清淀粉酶活性为157.1±23.7 U/l(P<0.001)。向使用传统葡萄糖溶液的患者血清样本中添加异麦芽糖糊精会导致淀粉酶活性呈剂量依赖性降低。检测结果表明ET7 - G7PNP与异麦芽糖糊精之间存在底物竞争。异麦芽糖糊精的AMAC荧光团染色及后续凝胶电泳未能证明异麦芽糖糊精与α -淀粉酶之间形成复合物。与淀粉酶的结果不同,异麦芽糖糊精不影响脂肪酶活性。

结论

目前的研究结果表明,异麦芽糖糊精在淀粉酶检测中作为底物发生竞争性相互作用。支持这一观点的是,SDS凝胶上的荧光团辅助寡糖电泳未能揭示“淀粉酶/异麦芽糖糊精复合物”的形成。脂肪酶检测应为诊断使用异麦芽糖糊精患者的胰腺炎提供一种替代且无干扰的方法。

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