1. School of Biology College of Science University of Tehran 2. School of Biology College of Science University of Tehran 3. School of Biology College of Science University of Tehran 4. School of Traditional Medicine Tehran University of Medical Sciences
Abstract
Introduction
Medicinal leech, during feeding, enter in host body many bioactive substances that often contain proteins and peptides. in regard to the therapeutic application of each of these proteins, leech therapies have extensive applications in medicine. in order to maximize the effect, it is important to select the suitable leeches that have the right amount of proteins of interest.
the aim of this study was found age-associated differences of saliva proteins in hirudo orientalis. the bradford method was used to determine total proteins and peptides concentration. sds-page 14% gel electrophoresis was performed for quantitative and qualitative changs of proteins from 5-245 kd.
Methods
For getting leech saliva, a solution of 0.001 m arginine in 0.9% nacl (phagostimulatory solution) at approximately 40° c fed to leeches and the solution that contained saliva collected in different ages. bradford method for protein and peptide concentration and sds-page 14% gel electrophoresis for protein detection was done.
Results
According to bradford absorption and 1d electrophoresis, quantitative and qualitative changes of the salivary proteins observed as with age passing, concentration and number of protein increased.for example hirudin, important protein in leech saliva, with nearly 7 kd increased with age passing.
Conclusion
This study was done according to wide therapeutic application of leech in medicine on hirudo orientalis salivary proteins in order to find protein changes in different ages. for this purpose phagostimulatory solution at approximately 40 ° c given to the leeches and collected solution with saliva. protein concentration was determined by bradford method. protein bands were also isolated by sds-page 14% gel electrophoresis between 5-245 kd. quantitative and qualitative differences were observed in the saliva in different ages, which generally increased with passing of time in different ages like hirudin. according to these results, select a suitable age with specified protein play an important role in the treatment.
