亚精胺对日本血吸虫成虫培养细胞SDH的影响  

THE EFFECTS OF SPERMIDINE ON SUCCINATE DEHYDROGENASE IN THE CULTURED CELLS FROM ADULT SCHISTOSOMA JAPONICUM

作　　者：易同寅[1] 董惠芬[1] 蒋明森[1] 明珍平[1] 钟沁萍[1] 

机构地区：[1]武汉大学医学院人体寄生虫学教研室,血吸虫病研究室,湖北武汉430071

出　　处：《中国寄生虫病防治杂志》2005年第5期339-341,T0001,共4页Chinese Journal of Parasitic Disease Control

基　　金：国家自然科学基金(No .30570243) ;湖北省卫生厅(No .JX1B067) ;武汉大学杰出青年基金( No .20030515) ;湖北省血吸虫病专家咨询委员会(No .301140165)资助项目。

摘　　要：目的以琥珀酸脱氢酶(SDH)为指标,探讨亚精胺(Spermidine ,Spd)对日本血吸虫成虫培养细胞生长代谢的影响。方法将24 d龄的日本血吸虫成虫用冷消化法制成细胞悬液,联合法将细胞接种于小盖玻片上培养。培养至第4 d ,一部分细胞用含Spd终浓度分别为0(对照)、25、50、75、80、90、100、150、200μmol/L的无血清培养基处理24 h,另一部分细胞用终浓度为75μmol/L的Spd分别处理0(对照)、12、24、36、48、60、72 h。然后用PBS清洗3次,再换用常规培养基继续培养。至第8 d ,对处理过的培养细胞进行SDH细胞化学染色,Olympus-BH2显微镜下观察、拍照,并将结果输入HPIAS-2000图像分析仪进行图像分析。结果用Spd作用24 h,日本血吸虫培养细胞的SDH活性随Spd浓度的升高逐渐增强,75μmol/L时达到最高,>75μmol/L,SDH活性逐渐减弱。当Spd作用浓度为75μmol/L时,日本血吸虫培养细胞的SDH活性随着Spd作用时间的延长逐渐增强,48 h时达到最高,然后逐渐减弱。统计学分析显示,Spd处理后的实验组培养细胞,其SDH活性与对照组细胞比较差异具有显著性(P<0 .05) ;用终浓度为75μmol/L的Spd处理培养细胞48 h,培养细胞的SDH活性显著强于其余各组(P<0 .01)。结论Spd可显著增强日本血吸虫培养细胞的生长代谢活性;用终浓度为75μmol/L的Spd处理48 h,培养细胞的SDH活性最强。Objective To investigate the effect of Spermidine （Spd） on succinate dehydrogenase （SDH） in the cultured cells from adult Schistosoma japonicum. Methods After the cells from 24-day old adult S. japonicum were inoculated on the small glass covered slips and cultured in medium RPMI-1640 containing 20% calf serum and a moderate amount of antibiotics for 4 days, they were divided into two groups. The cells in the first group were directly treated by Spd with concentrations of 0, 25,50,75,80,90, 150 and 200 μmol/L for 24 hours in medium RPMI 1640 with free serum, respectively. The cells in the second group were similarly treated by Spd of 75 μmol/L for 0, 12, 24, 36, 48, 60 and 72 hours, respectively. Then, they were moved into the original media. When they were cultured for 8 days, the cells were stained to show SDH by using a reformative method of Pearson. The stained cells were observed and photographed under a microscope （Olympus-BH2）. Simultaneity, the contents of SDH in the cultured cells were measured by the photometer-image analytical instrument （HPIA-2000） and analyzed statistically. Results The results showed that the activities of LDH in the cells from adult S. japonicurn treated by Spd of 75 μmol/L were stronger than those of other concentration when the cultured cells were directly treated by Spd with different concentrations for 24 hours（P〈0.01）. Similarly, the activities of SDH had a peak at 48 hours in the cells treated by Spd of 75 μmol/L for different time. There was significant difference between those treated by Spd and control （P〈0.01, P〈0.05）. Conclusion Spd could increase activities of LDH and growth of the cultured cells from S. japonicum. The activities of LDH were the strongest in the cultured cells treated by Spd of 75 μmol/L for 48 hours.

关 键 词：血吸虫 日本 培养细胞 亚精胺 琥珀酸脱氢酶(SDH) 

分 类 号：R383.24[医药卫生—医学寄生虫学]