Furlong S T, Samia J A, Rose R M, Fishman J A
Department of Medicine, Harvard Medical School, Boston, Massachusetts.
Antimicrob Agents Chemother. 1994 Nov;38(11):2534-40. doi: 10.1128/AAC.38.11.2534.
Although originally classified as a protozoan, Pneumocystis carinii is now considered to have fungal characteristics. Drugs typically used for the treatment of fungal infections target ergosterol. Because P. carinii is an important pathogen in AIDS and other immunocompromised patients, knowledge of the sterol content of this organism may be useful as a basis for developing new treatment strategies or for improving diagnosis. P. carinii organisms were harvested from infected rat lungs and were purified by filtration. Control preparations from uninfected animals were identically prepared. Lipids were extracted from the organisms and control preparations and were separated into neutral lipid, glycolipid, and phospholipid fractions by silicic acid chromatography. The neutral lipid fraction was further treated by alkaline hydrolysis and was analyzed by reversed-phase high-pressure liquid chromatography (HPLC), gas chromatography (GC), and GC-mass spectrometry (GC-MS). As shown by HPLC, the neutral lipid fraction from infected rats contained a minimum of six peaks, while in control preparations a single peak with a retention time identical to that of cholesterol was observed. The predominant sterol in these preparations was positively identified by GC-MS as cholesterol and constituted 80 to 90% of the total. The remaining peaks had relative retention times similar to those of phytosterols by both HPLC and GC, and the similarity of these sterols to those derived from plants and fungi was confirmed by MS. Ergosterol, however, was not present. These results provide further evidence for a close phylogenetic relationship between P. carinii and fungi and suggest that these sterols could be used as targets for drug development and for improving diagnosis.
尽管卡氏肺孢子虫最初被归类为原生动物,但现在被认为具有真菌特征。通常用于治疗真菌感染的药物靶向麦角固醇。由于卡氏肺孢子虫是艾滋病患者和其他免疫功能低下患者的重要病原体,了解这种生物体的固醇含量可能有助于开发新的治疗策略或改善诊断。从受感染的大鼠肺中采集卡氏肺孢子虫生物体,并通过过滤进行纯化。对未受感染动物的对照制剂进行相同的制备。从生物体和对照制剂中提取脂质,并通过硅酸色谱法将其分离为中性脂质、糖脂和磷脂部分。中性脂质部分进一步用碱性水解处理,并通过反相高压液相色谱(HPLC)、气相色谱(GC)和气相色谱-质谱联用(GC-MS)进行分析。如HPLC所示,感染大鼠的中性脂质部分至少含有六个峰,而在对照制剂中观察到一个保留时间与胆固醇相同的单峰。通过GC-MS确定这些制剂中的主要固醇为胆固醇,占总量的80%至90%。其余峰的相对保留时间通过HPLC和GC与植物固醇相似,并且通过质谱证实这些固醇与来自植物和真菌的固醇相似。然而,不存在麦角固醇。这些结果为卡氏肺孢子虫与真菌之间密切的系统发育关系提供了进一步的证据,并表明这些固醇可作为药物开发和改善诊断的靶点。