| Trichomoniasis is a common sexually transmitted disease, an estimated 170 million persons are infected with Trichomonas vaginalis worldwide. Metronidazole has been the principle medicine used to treat Trichomoniasis since 1960,the medicine is still effective now for approximately 90 to 95% of infections. However, metronidazole resistance have been developed in viro by growth in the presence of increasing concentrations of metronidazole and metronidazole treatment dose not cure all patients. In addition, the side effects such as nausea and gastrointestinal discomfort commonly reported and, with hypersensitivity reactions, can be severe enough to preclude the use of metronidazole for treatment some individuals. It is recently reported that tinidazole is efficacious against T. vaginalis isolates resistance to metronidazole at lower minimal lethal concentrations (MLCs) than metronidazole. Many traditional Chinese medicines have been used to treatment Trichomoniasis such as Sophora flavescens, Phellodendron amurense and Coptis deltoide. It was well known that the effects of the traditional Chinese medicines against T. vaginalis were less than metronidazole. T. vaginalis incubateded in vitro were used to screen medicines. T. vaginalis could grow in many culture medium which were cysteine- peptone-liver-maltose culture medium, Trypticase-Yeast Extract-Maltose culture medium and liver extract culture medium. However, roosevett park memorial institute culture medium 1640 (RPMI1640), which is popular culture medium for kinds of cell growth, had been used to incubate T. vaginalis as well. The suitable culture medium and accurate counting are essential to evalute the effect of drug compounds against T. vaginalis. The blood cell counting chamber assay and MTT assay had been used extensively counting the number of cell. The purpose of this study were to screen the suitable culture medium for T. vaginalis in vitro and determine effects of MSSZ compounds(the mixture of metronidazole, extracts of Sophora flavescens, extracts of Senecio scandens, Zedoray turmeric oil) on the growth of T. vaginalis in vitro. Methods: The suitable culture medium for the growth of T. vaginalis was screened from the rabbit liver extracts medium and RPMI-1640 medium. The proliferation of T. vaginalis was required exactly by comparing the two counting methods, the MTT stain and blood cell count methods. The normal ultrastructures of T. vaginalis were observed by transmission electron microscopes. Eleven medicines were investigated the effects on the culture organisms, which were metronidazole, extracts of S. flavescens, extracts of S. scandens, Z. turmeric Oil, and MSSZ compounds (the mixture of metronidazole and three extracts of the traditional Chinese medicines), and single compounds (the mixture of twins with the above simple medicines in equal proportion). The changes of ultrastructures of T. vaginalis treated with above medicines were observed by transmission electron microscopes. Results: The organisms of T. vaginalis could grow in the RPMI-1640 medicumat 37℃, and the optimal pH of 5.8. The organisms in the medium multiplied by 13.6 folds at 48h after inoculation, which could be used to evaluate the effect of trail medicines. The RPMI-1640 medium was a suitable medium to incubate the organisms of T. vaginalis, for its simple preparing procedure, and clear watch fields. The organisms in the liver extract culture medium multiplied by 26.4 folds at 72h after inoculation, the died liver cell and organisms of T. vaginalis were seen in watch fields. Some lively organisms of T. vaginalis were not stained with MTT. The results of trialed medicines on the T. vaginalis showed significant difference among MSSZ compounds groups, various single medicines groups (metronidazole, S. flavescens, S. scandens, Z. turmeric oil) and control groups (RPMI-1640 medium, 1% ethanol). The average of death rate of T. vaginalis treated by MSSZ compounds was 94.44% at the dilution of concentration ratio of 1:1 at 2h, all the organisms of T. vaginalis after... |
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