The Creation And Preliminary Practical Of Screening Platform Of Anti-hydatid Drugs In Vitro

Objective:To establish a drug screening platform used for observing the anti-Echinococcus granulosus activity of 100 kinds of harmine derivatives in vitro,hoping to find efficient anti-Echinococcus granulosus drugs.Methods:We obtained the echinococcus granulosus protoscoleces from slaughtering sheep liver cysts sterilely at the Xinjiang Hualing abattoirs.Before the experiment start,the protoscoleces will be cultured for a short time.Use hematoxylin-eosin stain to test the cytotoxicity of tween 80,dimethyl sulfoxide and and the mixtured solvent for the solution of above to E.granulosus protoscoleces which have a zero to ten days in vitro drug intervention experiment.Through high performance liquid chromatography(HPLC)to examine the solubilization affect of Tween 80 dimethyl sulfoxide and and the mixtured solvent for the solution of above to albendazole.Through HPLC and drug intervention experiments in vitro to examine the insecticidal effect of albendazole(ABZ),albendazole sulfoxide(ABZSO),harmine and total alkaloid of peganum harmala to select a positive control drug for the drug screening test.Use hematoxylin-eosin stain and drug intervention experiments in vitro to test the insecticidal effect of harmine derivatives on the basis of the above screening platform.Through observe anti-protoscoleces effect of harmine derivatives and compare the mice’s LD50 of harmine derivatives to select the harmine derivatives of high insecticidal effect and low toxicity.Use analysis of variance and Q test to analysis the result.Results:The cytotoxicity of three kinds of surfactants to protoscoleces enhanced with the concentration increasing.There is no obvious cytotoxic effect when protoscoleces had been interfered by low concentrations tween 80(0.05g/ml and 0.1g/ml),the same result also applies to low concentrations dimethyl sulfoxide(0.5g/ml and 1.0g/ml)and the mixtured solvent for the solution of above(P>0.05).The solubilization effect of three kinds of surfactants to albendazole increases with the concentration increasing.The solubility to albendazole of tween80(0.1g/100ml)dimethyl sulfoxide(1.0g/100ml)and the mixtured solvent for the solution of above are(7.47±0.11)μg/100ml,(0.77±0.15)μgg/100ml and(22.45±0.144)μg/100ml.The several times experiment results showed that the mortality of protoscleces with the drug of ABZ,ABZSO,harmine and total alkaloid of peganum harmala were(45±6.23)%,(20±4.38)%,100%and 100%in a ten days intervention experiment,but the anti-protoscoleces effect of ABZ and ABZSX had a significantly decrease when the time is two days.After a strict screening experiment,the result showed that the harmine derivatives with high anti-protoscoleces effect and low toxicity were DH-004,DH-109,DH-007 and DH-322.The LC50 and LD50 of them were(14.336±6.32)μg/ml,(19.23±2.12)μg/ml,(21.45±1.41)μg/ml and(31.21±2.21)pg/ml,ip250,ip 125,ip250 and ip500(P<0.05).Conclusion:This study established a drug screening platform used for observing the anti-Echinococcus granulosus activity of 100 kinds of harmine derivatives in vitro through selecting the cosolvent and positive control drug.In this screening platform,the cosolvent is 1%DMSO and the positive control drug is harmine.After screening experiment,the harmine derivatives of high anti-protoscoleces effect and low toxicity were DH-004,DH-109,DH-007 and DH-322.