ABSTRACT
The present study was designed to identify immunomodulatory components from the leech salivary gland of Haemadipsa sylvestris. The Sephadex G-50, Resource(TM) S column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC) were used to isolate and purify the salivary gland extracts (SGE). Structural analysis of isolated compounds was based on Edman degradation and matrix assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). The cDNA encoding the precursor of the compound was cloned from the cDNA library of the salivary gland of H. sylvestris. The levels of inflammatory mediators, including tumor necrosis factor-α (TNF-α), interferon γ (IFN-γ), interleukin-6 (IL-6), and monocyte chemotactic protein-1 (MCP-1) were assayed using an enzyme-linked immunosorbent assay (ELISA). The effects on cell proliferation and cell viability were observed using MTT assay. A novel neuropeptide Y (Neuropeptide Y-HS) from the leech salivary gland of H. sylvestris was purified and characterized. It was composed of 36 amino acid residues and the amino acid sequence was determined to be FLEPPERPAVFTSVEQMKSYIKALNDYYLLLGRPRF-NH2, containing an amidated C-terminus. It showed significant inhibitory effects on the production of inflammatory cytokines including TNF-α, IFN-γ, IL-6, and MCP-1. Neuropeptide Y was identified from leeches for the first time. The presence of neuropeptide Y-HS in leech salivary gland may help get blood meal from hosts and inhibit inflammation.
Subject(s)
Animals , Mice , Amino Acid Sequence , Immunologic Factors , Chemistry , Genetics , Inflammation , Drug Therapy , Allergy and Immunology , Interferon-gamma , Allergy and Immunology , Interleukin-6 , Allergy and Immunology , Leeches , Chemistry , Mass Spectrometry , Molecular Sequence Data , Neuropeptide Y , Chemistry , Genetics , Peptide Mapping , Salivary Glands , Chemistry , Tumor Necrosis Factor-alpha , Allergy and ImmunologyABSTRACT
Switching Mechanism At 5′ end of the RNA Transcript(SMART)is a technology used in biology researching,So far,there is no review only about SMART technology.So,the aim is to investigate the research developments of principles,methods and applications of the SMART technology.Based on some researches and combined with the review of the related literature at home and abroad,it analyzed and evaluated the latest development of the research on SMART.As the applications of the SMART technology expand in many fields day after day,it has been proved that the SMART technology is a very useful and efficient skill to construct full length cDNA library.As more and more researchers know this technology,the advantages of the SMART technology become obvious,and meanwhile the disadvantages of the SMART technology also show up.That is to say,the technology needed to be improved.
ABSTRACT
Objective To investigate the effect of ipratropi um bromide and atropine on airway mucin hypersecretion in the chronic bronchiti s model of rats. Methods The model was established by intratrac heal instillation of small dose of lipopolysaccaride (200 μg) ipratropium bromi de and atropine were administrated 3-21 d after the model was established. Patho logical changes, mucin in bronchoalveolar lavage fluid (BALF) and tracheal ring culture medium were examined before and after the adnsinistration of iprat ropium bromide and atropine. Results Characteristic pathologica l manifestions of chronic brochitis were found after instillation of LPS. Sig nificant decrease in the number of tracheal epithelia goblet cells, secretion of mucin were observed in ipratropium bromide and atropine treated rats. The eff ect of inhibiting the secretion of mucin of ipratropium bromide was higher than atropine. Conclusion It suggests that muscarinic acetylcholine receptor plays an important role in airway mucin hypersecretion in chronic bro nchitis model of rat. Ipratropium bromide exhibit a stronger inhibition effect on mucin hypersecretion than atropine, moreover no inhibition effect on mucocil iary clearance which was observed in atropine.
ABSTRACT
Objective To investigate the effect of ipratropi um bromide and atropine on airway mucin hypersecretion in the chronic bronchiti s model of rats. Methods The model was established by intratrac heal instillation of small dose of lipopolysaccaride (200 μg) ipratropium bromi de and atropine were administrated 3-21 d after the model was established. Patho logical changes, mucin in bronchoalveolar lavage fluid (BALF) and tracheal ring culture medium were examined before and after the adnsinistration of iprat ropium bromide and atropine. Results Characteristic pathologica l manifestions of chronic brochitis were found after instillation of LPS. Sig nificant decrease in the number of tracheal epithelia goblet cells, secretion of mucin were observed in ipratropium bromide and atropine treated rats. The eff ect of inhibiting the secretion of mucin of ipratropium bromide was higher than atropine. Conclusion It suggests that muscarinic acetylcholine receptor plays an important role in airway mucin hypersecretion in chronic bro nchitis model of rat. Ipratropium bromide exhibit a stronger inhibition effect on mucin hypersecretion than atropine, moreover no inhibition effect on mucocil iary clearance which was observed in atropine.