支原体感染对细胞的影响及四种药物对支原体的清除效果比较  

Effects of mycoplasma infection on cells culture and comparison with four drugs on mycoplasma

作　　者：庞白冰 楚元奎[1] 朱雅婷[1] 杨华[1] PANG Bai - bing CHU Yuan - kui ZHU Ya - ting YANG Hua(Ningxia Medical University ,Ningxia , Yinchuan ,Ningxia 750004, Chin)

机构地区：[1]宁夏医科大学临床医学院,宁夏银川750004

出　　处：《现代预防医学》2017年第17期3197-3202,共6页Modern Preventive Medicine

基　　金：国家自然科学基金项目(81460324)

摘　　要：目的探讨支原体感染对细胞的影响,并比较四种药物清除支原体的效果。方法使用Hoechst 33342荧光染色检测支原体污染情况,计数法检测细胞增殖,CCK-8实验检测细胞活力变化,qPCR检测细胞基因的表达,PCR检测培养细胞中支原体的感染状态,在感染的细胞中分别加入Plasmocin、Save It、阿奇霉素和环丙沙星并检测作用不同天数后支原体的清除情况。结果支原体感染后,乳腺癌MDA-MB-231和MCF-7细胞核周围异常荧光染色,细胞的形态发生明显改变;细胞计数及CCK-8实验结果显示支原体能够抑制MDA-MB-231和MCF-7细胞的增殖,并且细胞对CCK-8试剂的反应增加(P<0.001)。qPCR结果表明支原体感染对细胞基质金属蛋白酶(MMP)、甲酰肽受体(FPR)基因的表达有明显影响。对于感染了支原体的MDA-MB-231、MCF-7细胞和神经胶质瘤U87细胞,Plasmocin对MDA-MB-231细胞有效,Save It对三种细胞均有效,10μg/mL环丙沙星对MDA-MB-231、MCF-7细胞有效,10μg/mL的阿奇霉素对三种细胞均无效。除阿奇霉素外的三种清除支原体的药物均会影响细胞增殖。结论支原体感染会影响细胞的形态、增殖、活力以及基因表达,专门清除支原体的试剂Save It和Plasmocin有较好的清除效果。Objective To investigate the effect of mycoplasma infection on cells and compare the effects of four drugs on mycoplasma removal. Methods Hoechst 33 342 was used for fluorescence staining. Cell proliferation was measured by counting method. The cell viability was observed by the CCK - 8 test. The cell gene expression was measured by qPCR. The infection status of mycoplasma in culture cells was identified by PCR. SaveIt, Plasmocin, azithromycin and ciprofloxacin were used to treat mycoplasma infection,and the cell＇s infection status was detected after different days. Results Mycoplasma might cause breast cancer cells MDA -MB -231 and MCF -7 be stained abnormally fluorescence around the nucleus. Mycoplasma could change the morphological,inhibit proliferative ability and alter gene expression in the cells, and the response to the CCK - 8 reagent increased （P 〈 0.001 ）. For three cell lines infected with mycoplasma, Plasmocin was effective in MI）A -MB -231. Saveh was effective in all three. 10 μg/mL ciprofloxacin was effective in MDA - MB -231 and MCF -7 ,while 10 μg/mL azithromycin had no effect in all three cells. In addition to azithromycin, the other three drugs could affect cell proliferation. Conclusion Mycoplasma infection can affect the morphology, proliferation, viability and gene expression of cells. SaveIt and Plasmocin have a good effect to treat mycoplasma in the cell.

关 键 词：支原体感染 细胞培养 细胞增殖 防治 

分 类 号：R117[医药卫生—公共卫生与预防医学]