低分子透明质酸片段对红细胞聚集诱发作用及其种属特异性

The Inducing Effect of Low Molecular Hyaluronic Acid Fragments on Erythrocyte Aggregation and Its Species Specificity

旨在开发一种组织渗透性好的35 kDa透明质酸片段(HA35),探究HA35对红细胞的生物活性作用及其种属特异性。分别利用足量或轻度过量的重组人透明质酸酶PH20和牛提取透明质酸酶PH20,酶解切割平均分子量1600 kDa的透明质酸原料,制备一种能通过220 nm孔径的平均分子量35 kDa的透明质酸片段HA35,并进一步使用HA35与人和不同动物的红细胞共同孵育,研究HA35对红细胞的生物活性作用及其种属特异性。结果表明:HA35和其他平均分子量不同的透明质酸片段,通过CD44受体诱发人和多种实验动物的红细胞呈钱串状聚集,但存在种属特异性。HA35和其他平均分子量不同的透明质酸片段,诱发红细胞钱串状聚集的透明质酸片段的最小浓度与其分子量呈负相关,这种负相关关系首次被用来准确测定低分子透明质酸片段的分子量。HA35不仅诱发人和动物红细胞钱串状聚集,还诱发红细胞沉降率增加,被用于HA35产品的生产质控。

The aim of this study was to develop a 35 kDa hyaluronic acid fragment(HA35)with good tissue permeability,and to explore the biological activity and species specificity of HA35 on erythrocyte.In this experiment,sufficient or slightly excessive recombinant human PH20 hyaluronidase and bovine extract hyaluronidase PH20 were used respectively to cut the hyaluronic acid with an average molecular weight of 1600 kDa to prepare hyaluronic acid fragment HA35 with an average molecular weight of 35 kDa that could pass through a 220 nm pore size.And HA35 was incubated with human and different animal erythrocyte to study the bioactivity and species specificity of HA35 on erythrocyte.The results showed that HA35 and other hyaluronic acid fragments with different average molecular weight induced the erythrocytes rouleaux formation of human and various experimental animals by CD44 receptor,but there was species specificity.The minimum concentration of the erythrocytes rouleaux formation induced by HA35 and other hyaluronic acid fragments with different average molecular weight was negatively correlated with their molecular weight.This negative correlation was used for the first time to accurately determine the molecular weight of low molecular hyaluronic acid fragments.HA35 not only induces the erythrocytes rouleaux formation in human and animal,but also increases the erythrocyte sedimentation rate,which is used for production quality control of HA35 products.