MNNG对日本血吸虫成虫培养细胞糖类物质作用  

Effects of N-Methyl-N-Nitro-N-Nitrosoguanidine on carbohydrate in cultured cells from adult Schistosoma japanicum

作　　者：明珍平[1] 钟沁萍[1] 董惠芬[1] 朱俊勇[1] 蒋明森[1] 

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

出　　处：《中国公共卫生》2006年第1期50-52,共3页Chinese Journal of Public Health

基　　金：湖北省卫生厅基金资助(JX1B067);湖北省教育厅基金资助(2000B03014);武汉大学杰出青年人才基金资助

摘　　要：目的研究甲基硝基亚硝基胍（MNNG）对日本血吸虫成虫培养细胞糖类物质动态的影响。方法将日本血吸虫成虫细咆接种于小盖玻片上，置于RPMI-1640含20％小牛血清附加常量抗生素的常规培养基中培养。培养第4d，细胞被随机分为实验组和对照组。实验组细胞用含3μg／ml MNNG的常规培养基处理48h，对照组细胞用不含MNNG的常规培养基作相同处理。细胞经彻底清洗后．继续以常规培养基培养3周，然后换用含5％小牛血清的低血清培养基培养。NNNG处理后第1～8周．每周取实验组和对照组细胞进行高碘酸雪夫（PAS）染色和淀粉酶处理后的PAS染色，观察培养细胞内糖类物质的含量及分布变化。取培养第6周的染色细胞，用HPIAS-2000图像分析仪测定细胞内代表糖含量的吸光度，并作统计学分析。结果随着培养时间的延长。对照组培养细胞的着色逐渐变浅，糖含量逐渐减少；实验组细胞的着色则逐渐加深，糖类物质和糖原含量均增加，与对照组比较，差异有统计学意义（P〈0．01）。MNNG处理后第5周。实验组细胞着色最深．核质着色型第二类细胞和分裂细胞数目显著增加。结论MNNG诱导后，日本血吸虫成虫培养细咆内糖原和糖类物质含量均明显增加，分裂细胞增多。Objective To study the dynamic effects of N-Methy1-N-Nitrosoguanidine（MNNG） on carbohydrate in the cultured cells from adult Schistosoma japanicum. Methods The cultured cells from 32-day old S.japonicum were inoculated on cover slips and cultured in routine medium which was made up of RPMI-1640, 20% calf serum and moderate amount of antibiotics. At the 4th day, the cultured cells were divided into test group and control group at random. The cells in the test group were treated with MNNG which final concentration in the routine medium was 3μg/ml for 48 h, whereas those in the control group were treated with the routine medium free MNNG for the same time. After being washed thoroughly, they went on being cultured in the routine medium for 3 weeks; then were cultured in the low-serum medium which serum concentration was just 5 %. From the 1 st to 8 th week, the cells were stained weekly to show the content and distribution of hepatin and carbohydrate by Periodic Acid Scid Schiff reaction（PAS） and PAS staining after the cells were digested with amylase. The contents of carbohydrate in stained cells at the 6th week（the 5th week after inducement）were measured by using photometerimage analytical instrument（HPIAS-2000）and analyzed by statistic method. Results Along with the extending of the culture time, the staining of the cultured cells in the control group became weak, whereas those in the test group became strong. The contents of hepatin and carbohydrate of the cell in the test grop were significantly higher than those in the control group（P 〈 0.01）. At the 6th week, not only the staining of the cells treated by MNNG were the deepest, but also the 2nd type cells and dividing cells increased significantly. Conclusion MNNG could increase evidently the dividing cells and the content of hepatin and carbohydrate in the cultured cells from adult S. japonicum.

关 键 词：日本血吸虫 糖类 细胞化学 甲基硝基亚硝基胍(MNNG) 

分 类 号：A383.2[哲学宗教—马克思主义哲学]