Department of Chemistry, Lafayette College, Easton, PA 18042, USA.
Parasitol Res. 2010 Sep;107(4):947-53. doi: 10.1007/s00436-010-1959-y. Epub 2010 Jun 22.
The goal of this study was to characterize and quantify the various neutral and polar lipid classes in the BALB/c mouse that are associated with Echinostoma caproni infection. Ten infected mice and 10 uninfected control mice were used for this study (five infected and five uninfected were used for each of the neutral lipid and polar lipid studies). After 3 weeks postinfection, the mice were necropsied and various organs were removed and prepared for lipid class analysis. The organs used were liver, kidney, spleen, colon, cecum, anterior portion of the small intestine (SI), middle portion of the SI, and posterior portion of the SI. Lipids were determined by high-performance thin-layer chromatography (HPTLC) with Analtech 10 x 20 cm HPTLC-HLF silica gel plates. For neutral lipids, petroleum ether-diethyl ether-glacial acetic acid (80:20:1) mobile phase and 5% ethanolic phosphomolybdic acid detection reagent were used to determine the neutral lipids in each organ. Chloroform-methanol-deionized water (65:25:4) mobile phase and 10% cupric sulfate in 8% phosphoric acid detection reagent were used to determine the polar lipids in each organ. The analyzed polar lipids in all organs were phosphatidylcholine (PC), phosphatidylethanolamine (PE), and sphingomyelin (SM). Using HPTLC-densitometry for quantification, PC was found in the greatest amount and SM the smallest of all organs analyzed. The PE in the anterior portion of the SI was determined to be significantly greater (using the Student's t test with P < 0.05), with about twice the amount of PE in mice infected with E. caproni relative to the uninfected mice. No significant differences in any of the neutral lipid classes were found between infected and uninfected samples.
本研究的目的是描述和量化与棘口吸虫感染相关的 BALB/c 小鼠的各种中性和极性脂质类。本研究使用了 10 只感染的小鼠和 10 只未感染的对照小鼠(中性脂质和极性脂质研究各使用 5 只感染和 5 只未感染的小鼠)。感染后 3 周,对小鼠进行解剖,取出各种器官并准备进行脂质类分析。使用的器官有肝、肾、脾、结肠、盲肠、小肠前段(SI)、SI 中段和 SI 后段。通过高效薄层色谱法(HPTLC)用 Analtech 10 x 20 cm HPTLC-HLF 硅胶板测定脂质。对于中性脂质,使用石油醚-乙醚-冰醋酸(80:20:1)流动相和 5%乙醇磷钼酸检测试剂测定每个器官中的中性脂质。使用氯仿-甲醇-去离子水(65:25:4)流动相和 10%硫酸铜在 8%磷酸检测试剂中测定每个器官中的极性脂质。在所有器官中分析的极性脂质是磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)和神经鞘磷脂(SM)。使用 HPTLC-密度计进行定量分析,发现 PC 在所有分析器官中含量最高,SM 含量最低。SI 前段的 PE 被确定为显著增加(使用学生 t 检验,P < 0.05),与未感染棘口吸虫的小鼠相比,感染 E. caproni 的小鼠中 PE 的含量增加了约两倍。在感染和未感染样本之间未发现任何中性脂质类的显著差异。