异常黏液质证候大鼠阴茎组织差异性表达microRNA的筛选及生物信息学分析  

Screening of microRNAs profiles in rats model of abnormal phlegmatic syndrome and its medication disproof and bioinformatics analysis

作　　者：木萨.艾麦尔 蒋萍[2] 毛吾兰.买买提依明 马文静[4] 美合日古丽.萨塔尔 斯依提.阿木提 王天宇[1] 阿地力江.伊明 

机构地区：[1]新疆医科大学基础医学院人体解剖学教研室,乌鲁木齐830011 [2]新疆医科大学基础医学院生理学教研室,乌鲁木齐830011 [3]新疆医科大学基础医学院细胞生物学教研室,乌鲁木齐830011 [4]新疆医科大学中心实验室,乌鲁木齐830011

出　　处：《中国男科学杂志》2017年第1期3-8,共6页Chinese Journal of Andrology

基　　金：新疆维吾尔自治区自然科学基金联合基金项目(NO.2016D01C176);中国博士后科学基金第58批面上资助-西部地区博士后人才资助计划(2015M582773XB)

摘　　要：目的筛选异常黏液质证候及药物干预大鼠阴茎组织差异表达microRNA(miRNA),并验证miR-140-3p和miR-486两项关键指标,探讨其生物学意义。方法取80只性功能正常SD雄性大鼠,从中随机抽取10只大鼠设为正常对照组,余70只为造模组,用芫荽实+菠菜实+湿寒性环境的干预条件建立异常黏液质证候模型,通过生物学表征确认模型成功后,随机分为证候模型组、证候药物组。伊木萨克片干预3周后,提取阴茎组织总RNA进行miRNA Illumina高通量测序,筛选出异常黏液质证候及药物干预相关的候选差异表达miRNA,实时荧光定量PCR验证测序结果的可靠性,并进行生物信息学分析。结果筛选出证候相关miRNA候选标志物15个(上调13个,下调2个),伊木萨克片干预相关miRNA候选靶点标志物3个(均下调),验证结果显示miR-486在证候模型组较正常对照组表达升高(P<0.05),miR-486和miR-140-3p在证候药物干预组较证候模型组表达降低(P<0.05)。结论miR-486可作为异常黏液质证标志物,miR-140-3p、miR-486可作为伊木萨克片靶点标志物,但仍需大样本量验证。Objective To select the differently expressed microRNAs（miRNA） on abnormal Phlegmatic syndrome and it＇s medicine intervention rats＇s penile tissue, validate miR-486 and miR-140-3p, discuss the significance of biology of miR-486 and miR-140-3p. Methods Eighty healthy mature male SD rats were enrolled in this study, 10 of them were taken randomly as the control group, the other rats（70） were in the model group. Animal models of abnormal phlegmatic syndrome were established by treated with spinach and coriander diet in the cold humid environment. Biological characterization of rats were observed weekly until the models were successfully established. The rats were randomly divided into abnormal phlegmatic syndrome model group and abnormal phlegmatic syndrome model＇s medicine intervention group. Counterevidence was performed for 3 weeks, then penis tissue of the rats was collected to extract total RNA, and then subjected to Illumina Hiseq technology for screening differentially expressed miRNA, and then cluster analysis was performed, the candidate miRNA was selected and validated by qPCR. Results Total of 15 differentially expressed miRNAs related to Phlegmatic syndrome were identified , including 13 up-regulated miRNAs and 2 down-regulated miRNA; Three downregulation miRNAs related to target of medicine intervention were inversely regulated by Yimusake tablets treatment. The results of RT-qPCR showed that compare to control group, miR-486 was up-regulated in abnormal phlegmatic syndrome model group（P〈0.05）, miR-486 and miR-140-3p were down-regulated in abnormal phlegmatic syndrome model＇s medicine intervention group as compared to the abnormal phlegmatic syndrome model group（P〈0.05）. Conclusion miR-486 may be associated with the development of abnormal Phlegmatic syndrome. miR-140-3p, miR-486 may be as targets of Yimusake intervention.

关 键 词：异常黏液质证候 微RNAS 伊木萨克片 计算生物学 

分 类 号：R698.1[医药卫生—泌尿科学]