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首页> 外文期刊>Research communications in molecular pathology and pharmacology >Interaction of hyaluronic acid with mucin, evaluated by gel permeation chromatography.
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Interaction of hyaluronic acid with mucin, evaluated by gel permeation chromatography.

机译：透明质酸与粘蛋白的相互作用，通过凝胶渗透色谱法评估。

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摘要

Hyaluronic acid (HA) is known to increase the ocular bioavailability of ophthalmic drugs not only for its viscous properties but also for its specific affinity for ocular mucins. This phenomenon, called bio- or mucoadhesion, can be evaluated in vitro by mechanical tests which, however, require considerable amounts of mucin (M) that are difficult to obtain from ocular surfaces. Thus, we developed an alternative method, based on gel permeation liquid chromatography, to examine the interaction of HA with microgram quantities of mucin. HA (from human umbilical cord or rooster comb) were fractionated using a Sepharose CL-4B column, before and after incubation with porcine gastric mucin (PGM), and the fractions were analyzed by a specific assay based on the histological dye Stains-all. PGM interacted with high molecular weight (M.W). HA, causing the displacement of low M.W., non-covalently bound, HA fragments, which were eluted under a distinct chromatographic peak. By quantitating the relative area of this peak, an evaluation of the mucoadhesion of HA could be obtained. This method could be useful to study the interaction between HA and microgram quantities of ocular M (mucin), obtained from individual patients or normal subjects.
机译：透明质酸（HA）不仅可以提高眼科药物的眼部生物利用度，而且不仅具有粘性，还具有与眼粘蛋白的特异性亲和力。这种现象称为生物粘附或粘膜粘连，可以通过机械测试在体外进行评估，但是机械测试需要大量的粘蛋白（M），很难从眼表获得。因此，我们开发了一种基于凝胶渗透液相色谱的替代方法，以检查HA与微克量的粘蛋白的相互作用。在用猪胃粘蛋白（PGM）孵育之前和之后，使用Sepharose CL-4B色谱柱分离HA（来自人脐带或公鸡梳），并通过基于组织染料Stains-all的特定测定法分析这些级分。 PGM与高分子量（M.W）相互作用。 HA，导致低分子量，非共价结合的HA片段被置换，这些片段在不同的色谱峰下洗脱。通过定量该峰的相对面积，可以获得HA的粘膜粘附性的评估。该方法对于研究HA和从个体患者或正常受试者获得的微克量的眼M（粘蛋白）之间的相互作用可能是有用的。

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