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

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.