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目的培育恶性疟原虫萘酚喹抗性虫株,为恶性疟原虫抗性的深入研究提供实验虫株。方法采用恶性疟原虫FccSM/YN株,用Trager法进行体外连续培养,待其正常生长后,在培养基中间断添加不同浓度的萘酚喹进行克隆筛选,培育抗性,在培育前及用药后不同时段同步化处理疟原虫,使疟原虫小环状体率达95%以上,然后用Rieckmann体外微量法测定萘酚喹对培养虫株的半数抑制浓度(IC50)。结果药物刺激前(亲代)IC50为3.03 nmol/L;药物刺激后166 d IC50为43.07 nmol/L,为药物刺激前的14.22倍;停止药物刺激后25 d的IC50为18.98 nmol/L,较药物刺激后166 d下降55.94%。但仍然较亲代高6.26倍。结论连续体外培养药物间断刺激方法可以筛选培育出高度抗萘酚喹恶性疟原虫虫株。该株恶性疟原虫抗性不稳定,停药后抗性程度有所下降。
Objective To cultivate the paraquat resistant Plasmodium falciparum parahaemolyticus strains and provide an experimental strain for the further study on Plasmodium falciparum resistance. Methods The falciparum falciparum FccSM / YN strain was cultured in vitro by Trager method. After its normal growth, different concentrations of naphthoquine were intermittently added to the medium for clonal screening and resistance cultivation. Before and after incubation Plasmodium synchronously was treated at different time intervals to make the minocycline rate of Plasmodium more than 95%. The half inhibitory concentration (IC50) of naphthoquine on cultured strain was determined by Rieckmann in vitro micro-assay. Results IC50 was 3.03 nmol / L before drug stimulation and 43.07 nmol / L at 166 days after drug stimulation, which was 14.22 times higher than that before drug stimulation. IC50 was 18.98 nmol / L at 25 days after stopping drug stimulation, 166 d after stimulation decreased by 55.94%. But still 6.26 times higher than their parents. CONCLUSION: Continuous discontinuation of drug in vitro culture can be used to screen out highly resistant isolates of Plasmodium falciparum isolates. The strain of Plasmodium falciparum instability, after stopping the degree of resistance decreased.