Studies On Pharmacology And Purification And Properties Of Rana Temporaria

Frog that lives in all over the world except the Sourth Pole is a specieses ofamphibians. Antimicrobial peptides are a kind of important and small molecularimmune proteins in cell-free immunity of plants and animals. They have smallmolecular masses, good thermostability and antigenicity. They havebroadspectrum antimicrobial activity. Some of them are active against someviruses, fungi and cancer cell lines, but they are harmless to human normal cells.Recently, some reports on cancer cell lines make people more interested in them.Many scientists found that the polypeptide in the Rana Temporaria have a lot ofanti-inflammatory and analgesic activities.It is possible that antimicrobial peptidesmay become a kind of new antimicrobial and antitumoral drug. In a word,antimicrobial peptides would greatly attribute to the promotion of people's healthlevel in the near future.Boman, a science of Sweden, established the existence of antimicrobialpeptides-Cecropins in 1975. Subsequently, many antimicrobial peptides arediscovered, including bacteria, fungi, insects, amphibians and mammals etc. Theyall produce antimicrobial peptides. But every animal has different peptides, suchas Magainins, Protegrins,Melitins, Defensins. And the endogenous antimicrobialpeptides of plants and animals are typically amphipathic molecules composed of12 to 45 amino acid residues. These peptides may be produced constitutively oronly after infection or injury, and they often contain excess lysine and arginineresidues.According to their characters, they were divided into four groups. 1) Thececropins. They are 4KDa peptides and active on both Gram-negative andGram-positive cells and devoid of cysteines and have the structure of twoamphipathic α -helices. 2) The insect defensins. They are 4KDaanti-Gram-positive peptides with six cysteines engaged in three intramoleculardisulfide bridges. 3) Small, proline-rich peptides. They are active against Gram-negativebacteria and have been isolated from Hymen-opera, Diptera andHemoptera. A remarkable feature of the peptides is the presence of anOglycosylated substitution. 4) Glycine-rich polypeptides. The molecular weightsof the peptides range from 10 –30KDa. They are active essentially onGram-negative bacteria.Gel is new preparation in recent years to meet the need of clinic. It use somenew macromolecular medical materials, such as Carbomer, CMC-Na, PVP, PVAect. It can improve the part concentration and prolong the release and pervasion. Ithas the advantages of stabilities and little badness response.In this article we established the quality standard of Rana Temporaria.We started our work in order to find new activion of antimicrobial peptides.The anti-inflammatory and analgesic properties of the extract of the skin of Ranatemporaria were investigated in the present study. The tests of subplantar oedemainduced by carrageenan model,ear oedema induced by dimethylbenzene modelshowed strong anti-inflammatory.The results of mouse writhing test and hot plate test,tail flick tests showedthat the analgesic peptides had strong inhibitive effect on visceraglgia andsomatalgia。From the overall results we can concluded that the tested the big doseshowed a significant analgesic activity, while only the small dose showed a loweranalgesic activity. The analgesic chemistry substance of Rana Temporaria issimilar to Ibuprofen in analgesic strength.In this article, anti-inflammatory and analgesic peptides were isolated andpurified from the s Rana Temporaria . Its physicochemical properties andpharmacological effect were studied.The purification procedure included gelfiltration chromatography and sds-page chromatography . Pharmacology testsdemonstrated that peak 2 have analgesic activity. Their homogeneity weredemonstrated by high performance liquid chromatography.Their molecularweights were 8000 and 24000 Dr calculated from measurement inSDS-polyacrylamide gel electrophoresis.