重组水蛭素-2在家兔鼻黏膜中降解的体外实验

目的考察重组水蛭素-2(rHV2)在家兔鼻黏膜中的降解情况。方法建立用HPLC测定家兔鼻黏膜和肠黏膜组织匀浆液中rHV2浓度的方法；测定不同浓度rHV2在家兔鼻黏膜组织匀浆液中及rHV2在不同pH值的家兔鼻黏膜组织匀浆液中的降解；比较家兔鼻黏膜和肠黏膜组织匀浆液对rHV2的降解；考察rHV2在家兔鼻黏膜酶提取液中的稳定性以及加入酶抑制剂对rHV2在家兔鼻黏膜组织匀浆液降解的抑制作用。结果rHV2在鼻黏膜匀浆液中的降解具有明显的药物浓度和pH依赖性；rHV2在鼻黏膜匀浆液中的稳定性优于在肠黏膜匀浆液中的稳定性；鼻黏膜酶提取液对rHV2的降解作用较小；酶抑制剂杆菌肽对rHV2在鼻黏膜匀浆液中的降解具有一定的抑制作用。结论与口服给药途径相比，rHV2鼻腔给药更有优势。

Degradation of recombinant hirudin-2 in nasal mucosa of rabbits in vitro

Aim To investigate the degradation of recombinant hirudin-2(rHV2) in nasal mucosa of rabbit.Methods The specific and accurate HPLC method was developed for analyzing rHV2;The degrading ratios of rHV2 at different concentrations and at pH conditions in rabbit nasal mucosa homogenate were determined;The results in nasal mucosa homogenate were compared with that in small intestinal mucosa homogenate of rabbits.The stability of rHV2 in the enzyme extract of nasal mucosa surface and the effect of proteolysis inhibitor bacitracin on the degradation of rHV2 in nasal mucosa homogenate were also estimated.Results The degradation of rHV2 in rabbit nasal mucosa homogenate showed concentration-and pH-dependence;rHV2 in nasal mucosa homogenate was more stable than that in intestinal mucosa homogenate.Also rHV2 was more stable in the enzyme extracts of nasal serosal surface than that of mucosa surface.Addition of bacitracin was able to inhibit the degradation to some degree.Conclusion Comparing with oral administration,rHV2 nasal delivery was a more tolerant route.