应用微阵列芯片分析草酸钙结晶肾损伤中LncRNA表达谱的差异  

Microarray Analysis of Kidney Injury by Crystallization of Calcium Oxalate Differences in LncRNA Expression Profiles

作　　者：张彩虹[1] 谌卫[1] 袁继行 张杰[1] 马文杰[1] 刘敏[3] 王田田[2] 蔡杰[4] 郭志勇[1] 

机构地区：[1]第二军医大学附属长海医院肾内科,上海200433 [2]第二军医大学遗传研究所,上海200433 [3]第二军医大学附属长海医院泌尿外科,上海200433 [4]第二军医大学附属东方肝胆医院肝外三科,上海200433

出　　处：《中国中西医结合肾病杂志》2016年第1期14-17,I0002,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金资助项目(No.81270773;81573759);上海市科委中药支撑计划项目(No.13401900105);上海市教委科研创新重点项目(No.14ZZ080)

摘　　要：目的:利用微阵列芯片对结晶组小鼠和正常组小鼠的基因表达谱进行比对分析,筛选出差异基因,探讨lncRNA与肾结石的关系。方法:将6只C57BL/6雄性小鼠随机分为结晶组和正常组,两组腹腔分别注射50 mg/kg生理盐水和100 mg/kg乙醛酸盐。7 d处死全部小鼠,对两组小鼠的肾组织,提取总RNA后进行差异lncRNA的筛选、聚类分析以及小鼠lncRNA与人同源性分析,采用实时定量RT-PCR验证人鼠同源性lncRNA。结果:芯片结果显示差异表达lncRNA共376个,其中结晶组表达上调154个,下调222个;差异表达mRNA共3 062条,上调1 914条,下调1 148条。结论:Sprr2c、CHCHD4P4可能参与肾结石发生的调控。Objective： To detect the expression of lncRNA in normal and crystallized kidney tissue using microarray,and to discuss the relationship between lncRNA and kidney stone.Methods： Six male C57BL /6 mice were randomly divided into two groups：normal saline control group and glyoxylate experiment group.Two groups were injected with 50 mg / kg NS,100 mg / kg glyoxylate into abdominal cavity.All mice were killed and their kidneys were harvested to make microarray.LncRNA identification were performed by microarray of lncRNA hybridization and cluster analysis was used with Map Splice（ v2.1.6） software.We applied the blast method towards the mouse lncRNA to human lncRNA to discover the homologous lncRNA.Homologous lncRNA and their mouse lncRNA were confirmed by real time quantification RT-PCR with RNA-tailing and primer extension.Results： Totally 376 different lncRNAs and3062 different mRNAs were found expressed in crystal group,with 154 of lncRNAs expression were up-regulated and 222 of lncRNAs expression were down-regulated.Conclusion： Sprr2c、CHCHD4P4 may play an important role in the pathogenesis and development of kidney stone.

关 键 词：草酸钙结晶 长链非编码RNA 人近曲肾小管上皮细胞(HK-2) 微阵列芯片 

分 类 号：R692.4[医药卫生—泌尿科学]