摘要
目的探讨靶向白色假丝酵母菌分泌型天冬氨酸蛋白酶(SAP)的小分子干扰RNA(siRNA)对白色假丝酵母菌SAP的基因表达影响,以期为RNA干扰技术抑制真菌毒力提供实验基础。方法设计并合成SAP靶向的siRNA,用醋酸锂转染白色假丝酵母菌,试验设空白对照组、错义对照组、SAP siRNA组,采用RT-PCR测定转染后白色假丝酵母菌SAP mRNA表达水平,制备BSA培养基测定天冬氨酸蛋白酶活性及小鼠毒力实验。结果与空白对照组、错义对照组相比,转染48h后,siRNA组白色假丝酵母菌中SAP mRNA表达明显下调为(0.51±0.05),siRNA组天冬氨酸蛋白酶活性PA值为(0.84±0.04)均高于空白及错义对照组,小鼠30d死亡率显著降低为20.0%。结论化学合成的靶向SAP siRNA能够下调白色假丝酵母菌中SAP基因的表达,降低白色假丝酵母菌的毒力,该实验为进一步研究应用RNA干扰技术抗真菌治疗奠定基础。
OBJECTIVE To explore the effect of targeting Candida albicans siRNA on the SAP gene expression. and to provide an experimental basis for RNA interference to inhibit fungal virulence., METHODS The SAP targeting siRNA was designed and synthesized. and transfected into C. albicans by lithium acetate. In the experiment. there were blank control group. the missense control group and SAP siRNA group. the SAP mRNA expression levels were detected by RT-PCR after transfection C. albicans , and mouse virulence test and aspartic protease activity were measured by preparation of BSA medium. RESULTS Compared to the control group and the missense control group. after transfection for 48h. the C. albicans SAP mRNA expression in siRNA group was significantly down-regulated to 0. 51 ± 0. 05 , and the aspartic protease activity P A value was significantly higher （0. 84 ± 0. 04） than that in the other two groups. the 30-day mortality in mice was significantly reduced to 20. 0 %. CONCLUSION Chemical synthetic targeting SAP siRNA can reduce the SAP gene expression in the C. albicans and the virulence of C. albicans. The experiment has laid the basis for further research and application of RNA interference technology antifungal therapy.
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2014年第5期1044-1046,共3页
Chinese Journal of Nosocomiology
基金
贵州省科技厅基金资助项目(2273)
