五种非常规使用化学物抗新型隐球菌体外药敏实验研究

In vitro drug sensitivity of five unconventional chemicals against cryptococcus neoformans

目的研究五种目前临床非常规使用化学物的体外抗隐球菌活性,为隐球菌病的治疗增加新型药物的选择。方法选择2018~2021年本院分离得到的10株临床不同来源的新型隐球菌分离株作为实验菌株;用基质辅助激光解吸离子飞行质谱技术(MALDI-TOF-MS)进行菌种的鉴定;采用微量肉汤稀释法以氟康唑为参考药物对五种非常规药物雷夫康唑(RFC)、盐酸舍曲林(SRT)、小檗碱(BRB)、香芹酚(CRV)、布洛芬钠(IBU)药物分别测定抑制90%菌株生长的最低药物浓度(MIC_(90))及棋盘微量肉汤稀释法与氟康唑(FCA)进行两两组合或临床可能使用的其他两种药物组合进行联合药敏实验测定MIC_(90),并计算部分抑制浓度指数(FICI)。结果雷夫康唑、盐酸舍曲林、小檗碱、香芹酚、布洛芬钠药物的MIC_(90)分别为≤0.03μg/ml、4~8μg/ml、8~32μg/ml、128~512μg/ml、2~8 mg/ml;联合药敏显示各双药组合的FIC指数分别如下:SRT与FCA为0.38~1;SRT与BRB为0.53~1.13;BRB与FCA为0.31~0.75;SRT与IBU为0.63~1.50;FCA与IBU为2.13~4.13。结论五种试验药物体外均具有一定的抗隐球菌活性,尤其是雷夫康唑具有极低的MIC_(90)。舍曲林、小檗碱也具有较低的MIC_(90)值,体外活性接近氟康唑。香芹酚、布洛芬钠体外抗隐球菌活性相对有限。联合药敏实验中,除了氟康唑与布洛芬钠显示体外拮抗作用外,其他不同药物组合之间基本表现为相加或无关作用。

Objective To study the several current clinical unconventional use of chemicals in vitro activity against cryptococcus neoformans,thus to increase new drugs selection for the treatment of cryptococcosis.Methods Ten clinical strains of cryptococcus neoformans isolated from different sources in our hospital from 2018 to 2021 were selected as experimental strains.Matrix assisted laser desorption ion flight mass spectrometry(MALDI-TOF-MS)was used to identify.The lowest concentration of a drug that inhibits the growth of 90%of pathogens(MIC_(90))of five non-conventional drugs(RFC),sertraline hydrochloride(SRT),berberine(BRB),carvaxol(CRV)and ibuprofen sodium(IBU)were determined by microbroth dilution method with fluconazole as reference drug,and the checkerboard microbroth dilution method was used to pair fluconazole(FCA)combination susceptibility tests were performed to determine MIC_(90)and calculate partial inhibitory concentration index(FICI)for combination or other two drug combinations that may be clinically used.Results The MIC_(90)of ravaconazole,sertraline hydrochloride,berberine,carvacrol and ibuprofen were≤0.03μg/ml,4-8μg/ml,8-32μg/ml,128-512μg/ml,and 2-8 mg/ml respectively.The FIC index of each drug is shown as follows:sertraline hydrochloride and fluconazole were 0.38-1,sertraline hydrochloride and berberine were 0.53-1.13,berberine and fluconazole were 0.31-0.75,sertraline hydrochloride and ibuprofen sodium were 0.63-1.50,fluconazole and ibuprofen sodium were 2.13-4.13.Conclusion Several experimental drugs all have certain anti-cryptococcus activity in vitro,especially ravaconazole has very low MIC_(90),sertraline and berberine also have low MIC_(90).In vitro activity the both drugs was close to fluconazole.Ibuprofen sodium and carvacrol had limited activity against cryptococcus in vitro.Combined drug sensitivity test showed that fluconazole and ibuprofen sodium showed antagonistic effect in vitro,other drug combinations showed additive or irrelevant effect.Combined with the pharmacological advantages of each drug,this study will provide some laboratory basis for the selection of new drugs for internal and external use in clinical anti-cryptococcus.