Isolation and purification of agkistrodon venom defibrase and experimental study of its transrectal dosing
OBJECTIVE To isolate and purify defibrase from agkistrodon venom by chromatography and explore the feasibil-ity of transrectal dosing into blood circulation to reduce the level of fibrinogen in vivo.METHODS Defibrase was isolated from crude venom of Agus agicoides.After a transrectal injection,the distribution of defibrase in the organs of mice was observed by animal imaging.The effect of defibrase on tissues was detected by pathology examination.RESULTS Electrophoretically pure defibrillinase was acquired by isolation and purification of crude venom of sharp-nosed pit viper.It was safe after transrectal dosing without acute toxicity or obvious pathological changes in rectal tissues.Immunohistochemistry indicated that fibrinogenin was dis-tributed in rectal tissues(P＜o.01).Shotgun mass spectrometry was utilized for measuring the plasma level of fibrinogen frag-ments after 1 h of dosing.Animal imaging revealed that dye-labeled fibrinogen was predominantly distributed in cecal tissues after 3 h of dosing with a gradual decrease.Rectal administration of fibrinogen reduced fibrinogen level in vivo(P＜0.01).CONCLUSION Electrophoretic pure defibrase is acquired from crude venom of agkistrodon by two-step chromatography.Defi-brase may be absorbed into blood circulation and reduce fibrinogen level after transrectal dosing.
defibrasevenom of Agkistrodon acutusrectal dosingseparation and purification
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