羊瘙痒因子感染小鼠脑组织中MicroRNA—375对PDK1蛋白表达调控研究

Study on the regulation of microRNA-375 on PDK1 expression in brain tissue of scrapie agent 139 A infected mice

目的 研究羊瘙痒因子139 A感染小鼠脑组织中3＇磷酸肌醇依赖的激酶1（3＇-phosphoinositide-dependert kinase 1,PDK1）和micro RNA-375（miR-375）含量的变化;探讨miR-375在朊病毒病发生发展过程中对PDK1蛋白表达的影响.方法 以羊瘙痒因子139 A感染的终末期小鼠脑组织为研究对象,通过Western blot方法检测脑组织中PDK1蛋白的含量;采用RT-PCR和二代测序的方法检测脑组织中miR-375含量的变化;利用了PmiR-REPORT报道系统检测miR-375对PDK1 3＇UTR调控的影响.结果 PDK1在羊瘙痒因子139 A感染小鼠脑组织中的表达量相对于正常对照组增高6倍左右,而miR-375表达量降低（P＜0.05）;PmiR-REPORT报道系统结果表明PDK1 3＇UTR是miR-375的调控靶点.结论 在羊瘙痒因子139 A感染小鼠终末期脑组织中PDK1的含量明显增加,可能与miR-375对PDK1 3＇UTR的调控有关.

Objective To investigate the regulation of microRNA-375 miR-375） on the expressions of 3＇-phosphoinositide-dependent kinase 1 （PDK1） in the brain tissues of scrapie agent 139 A infected mice.Methods PDK1 protein in 139 A infected mice brain tissue was detected by WB and immunochemistry.The change of microRNA-375 was detected by reverse transcription polymerase chain reaction （RT-PCR） and next-generation sequencing method.The pmiR-REPORT reporter system was used to value the regulation of miR-375 on PDK1 3＇-untranslated region （3＇UTR）.Results The expression of PDK1 in the brain tissue of 139 A infected mice was significantly increased as compared to that of control group,while the expression of miR-375 was decreased.The result of pmiR-REPORT reporter system showed that PDK1 3＇UTR was the regulation target of miR-375.Conclusions The expression of PDK1 in the brain tissue of 139 A infected mice was significantly increased,which was probably related to the regulation of miR-375 on the 3＇UTR of PDK1.