亚精胺对日本血吸虫成虫培养细胞核仁组织区相关嗜银蛋白的影响  

Effects of spermidine on argyrophilic nucleolar organizer region associated proteins in cultured cells from adult Schistosoma japonicum

作　　者：陈喜珪[1,2] 易同寅[1] 明珍平[1] 钟沁萍[1] 蒋明森[1] 董惠芬[1] 

机构地区：[1]武汉大学基础医学院人体寄生虫学教研室,血吸虫病研究室,咸宁学院医学院430071 [2]咸宁学院医学院

出　　处：《中国地方病学杂志》2006年第1期50-53,共4页Chinese Jouranl of Endemiology

基　　金：国家自然科学基金资助项目(30570243);湖北省卫生厅基金资助项目(JX1B067);湖北省血吸虫病专家咨询委员会资助项目(301140165);武汉大学医学院杰出青年基金资助项目(2004)

摘　　要：目的以日本血吸虫培养细胞核仁组织区相关嗜银蛋白(Argyrophilicnucleolarorganizerregionassociatedproteins,AgNORs)为评价指标,探讨亚精胺(Spermidine,Spd)对培养细胞的促增殖作用。方法采用联合法将虫龄为24d的日本血吸虫成虫细胞接种于小盖玻片上,培养于常规培养基中。接种后第3天,随机分为实验组和对照组。将实验组细胞用无血清培养基配制的终浓度为75μmol/L的Spd处理48h,对照组细胞则用不含Spd的无血清培养基处理,时间相同;PBS清洗3次后换用常规培养基继续培养。于第7、14、21、28、35天分别对培养细胞进行AgNORs染色。使用HPIAS-2000图像分析系统,计算培养细胞核内AgNORs颗粒数及平均吸光值,并作统计分析。结果经Spd处理,各实验组培养细胞AgNORs着色明显深于对照组;但随着培养时间的延长,实验组与对照组细胞着色均逐渐变浅。各实验组细胞AgNORs颗粒数目与平均吸光值高于对照组,二者比较差异有统计学意义(u7=50.95、43.78;u14=55.23、43.54;u21=36.65、32.25;u28=31.6、32.95;u35=28.86、9.54;P<0.01)。各实验组AgNORs吸光值两两比较,除第7天与第14天之间差异无统计学意义外(q=0.008058,P>0.05),其余差异都有统计学意义(P<0.01)。结论Spd具有促进日本血吸虫培养细胞增殖的能力,每周用Spd处理培养细胞1次,能更大限度地促进血吸虫培养细胞的生长。Objective To investigate the effect of spermidine （Spd） on the contents of argyrophilic nucleolar organizer region associated proteins （AgNORs） in the cultured cells and proliferation of the cultured cells from Schistosomajaponicum（S. japonicum）. Methods The cells from S. japonicum were inoculated on the small glass cover slips. The cultured cells were randomly divided into test and control groups after they were cultured for 3 days in routine media. The cultured cells in the test group were treated with Spd at a concentration of 75 μm/L for 48 hours in the media without serum. Then, the cells were cultured in routine media. The cells in the control group were treated similarly to those in the test group except they were free from Spd in the media. The cells were stained to show contents of AgNORs by the silver colloid technique from 1st to 5th weeks. Such two parameters AgNORs as number of granules and average optical density were measured by using the photometer-image analytical instrument （HPIA-2000） and analyzed statistically. Results After treated by Spd, the AgNORs staining in the cultured cells of test groups were deeper than those of control groups. Along with the prolongation of cultural time, those of test and control groups were all got weaker gradually. The number of granules and average optical density, were higher in the test group than those in control. The contents of AgNORs in the cultured cells had significant difference between test and control groups （u7 = 50.95,43.78; U14 = 55.23,43.54; U21 = 36.65,32.25; u28 = 31.6,32.95; u35 = 28.86,9.54;P 〈 0.01 ）, respectively. Among the test groups, there was significant difference average optical density except for that in the cells cultured for 7 days and 14 days. Condusions Spd promote proliferation of the cultured cells from S. japonicum. If the cultured cells were treated by Spd once every week, the growth of the cultured cells could be going to be better.

关 键 词：亚精胺 血吸虫 日本 细胞 培养的 核仁组织区相关嗜银蛋白 

分 类 号：R383.24[医药卫生—医学寄生虫学] R971.2[医药卫生—基础医学]