Flowmetric study with hyaluronidase and chondroitin sulfate demonstrates the partaking of endothelial glycocalyx in microcirculation disturbances

Abstract

Modification of bovine testicular hyaluronidase with chondroitin sulfate altered on the contrary the
glycation character of native and modified enzyme with neutral and charged saccharide derivatives. Monoand di- saccharides inactivated more the native enzyme than modified biocatalyst. The glycation with
N-acethylhexosamines showed the counter picture: modified enzyme has been inactivated more then native
hyaluronidase. The reason of observed effect was related with interactions between N-acethylhexosamine (with
reducing end) glycation agents, on the one hand, and modified hyaluronidase with altered surface electrostatic
potential due to modification with chondroitin sulfate, on the other hand. These properties of hyaluronidase
derivatives gives us the informative research enzyme test for determination in vivo the dominant glycation
agents in bloodstream and their origin. The interaction of hyaluronidase derivatives with hyaluronan fragments
and their mixture has confirmed the opportunity of such evaluation. On the rat model of post ischemic perfusion
of limb we evaluated the restoration of microcirculation level with help of laser doppler flowmetry method
with application of enzyme derivatives and their components. After preventive administration the native
hyaluronidase accelerate the microcirculation restoration in post ischemic period of rat limb. Modified enzyme
was inhibited with glycocalyx degradation products (with N-acethylhexosamines at reducing end) pronouncedly.
The results of laser doppler flowmetry (after different schemes of assayed substance administration) warranted
the participation of endothelial glycocalyx in microcirculation disturbances.