肝基质与MNNG对日本血吸虫成虫培养细胞糖细胞化学作用的研究  

Effects of the combined use of liver matrix and N-methyl-N-nitro-N-nitrosoguanidine(MNNG) on the content and distribution of carbohydrates in the cultured cells from adult Schistosoma japonicum

作　　者：明珍平[1] 钟沁萍[1] 董惠芬[1] 蒋明森[1] 张兆仁 

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,武汉430071 [2]湖北省武汉市血吸虫病防治办公室

出　　处：《中国人兽共患病学报》2006年第3期259-262,共4页Chinese Journal of Zoonoses

基　　金：湖北省自然科学基金(2000J075);湖北省;武汉市血吸虫病专家咨询委员会;武汉大学杰出青年人才基金资助

摘　　要：目的研究肝基质与甲基硝基亚硝基胍（MNNG）对日本血吸虫成虫培养细胞糖类物质的影响。方法日本血吸虫成虫细胞分别接种于普通小盖玻片（实验1组）和铺敷有肝基质的小盖玻片上（随机分为实验2组和3组），培养于含20％小牛血清及常量抗生素的RPMI-1640常规培养基中。培养第4d，实验1组和3组细胞分别在含3μg/ml MNNG的常规培养基中培养48h，彻底清洗后继续用常规培养基培养；实验2组细胞用不含MNNG的常规培养基处理相同时间。培养第4w。均换用含5％小牛血清的低血清培养基培养。培养第5～7w，每周取各组细胞进行高碘酸雪夫（PAS）染色和唾液淀粉酶处理后的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6w（MNNG作用后第5w）的各组染色细胞，用HPIAS-2000图像分析仪测定细胞内代表糖含量的光密度，并作统计学分析。结果实验3组细胞与实验1、2组比较，PAS染色显示的四种类型细胞其着色均显著增强，淀粉酶处理后的PAS染色显示细胞内糖原含量增多，差异均具显著性（P〈0．01）。培养过程中，第二类细胞数目逐渐增多，体积增大；分裂细胞增多。这种状况在MNNG处理后第5w最为明显。结论肝基质和MNNG联合可显著增强日本血吸虫培养细胞的糖代谢和分裂增殖能力，二者具协同作用能力。To observe the effects of the bio-matrix from liver of rabbits （liver matrix） and N-methyl-N-nitro-N-nitrosoguani- dine （MNNG） on the content and distribution of carbohydrates in the cultured cells from adult Schistosoma japonicum, the cultured cells from 32 days old S. japonicum were inoculated on ordinary cover slips （control group 1） or cover slips smeared with liver matrix （randomly divided into experimental group 2 and 3）, and cultivated in routine RPM I -1640 medium containing 20 % calf serum and moderate amount of antibiotics. At the 4 th day of cultivation, cultivated cells of group 1 and 3 were induced with routine medium containing MNNG in concentration of 3 μg/ml for 48 hrs, whereas those in group 2 were just treated with the routine medium at the same time. Four weeks later, the cultured cells were thoroughly washed and still cultivated in low serum medium containing 5 % calf serum. At 5-7 th week, the cultured cells were stained with Periodic Acid Schiff （PAS）stain and the mucus amylase-treated PAS staining to demonstrate the content and distribution of carbohydrates in the cultured cells. The content of carbohydrates in the stained cells at the 6 th week （5 th week after induction） were measured by using photometerimage analytical instrument （HPIAS-2000） and analyzed by statistic methods. The experimental results showed that in comparision with the cells in group 1 and 2, the cultured cells in group 3 showed an increase in intensity of PAS staining in 4 types of cells, and the PAS staining treated with amylase demonstrated an increase of the cellular hepatin content with significant differences. In the course of cultivation, the volume of the second type of cells in experimental group 3 enlarged gradually and the number of the dividing cells increased, especially at the 5 th week when the cultured cells were induced with MNNG. It is evident that liver matrix and MNNG show cooperative effect on the cultured cells, in which they can enhance the carbohydrate metabolism and

关 键 词：日本血吸虫 培养细胞 糖 肝基质 甲基硝基亚硝基胍 

分 类 号：R383.2[医药卫生—医学寄生虫学]