长链非编码RNA-uc.412调控系膜细胞增殖的作用及机制研究

LncRNA uc.412 was found up-regulated in hyperplastic mesangial cells by high-throughput lncRNA microarray. In previous study, uc.412 over-expression could enhance proliferation ability of mesangial cells; bio-informatic analysis and data showed that AATF might be the target gene of uc.412; AATF was found overexpressed in renal biopsy tissues of children with mesangial proliferative glomerulonephritis (MsPGN), and AATF could stimulate mesangial cells proliferation. Based on the previous data, we assumed that uc.412 could regulate the proliferation ability of mesangial cells through AATF. In order to identify this hypothesis, we would explore the expression character of uc.412 in children glomerulonephritis; asses the function of uc.412 in mesangial cells proliferation process by overexpression and silence methods; clarify the undergoing mechanism of uc.412 regulating mesangial cells proliferation through AATF. This study might be helpful to understand the impact of uc.412 in mesangial cells proliferation, and afford potential therapeutic targets for children with MsPGN.

uc.412是我们应用高通量lncRNA芯片筛选获得的、在异常增生系膜细胞中表达显著上调的lncRNA。前期证实，uc.412过表达显著增强系膜细胞的增殖活力；生物信息学分析及初步实验结果表明AATF可能是uc.412的下游靶基因；AATF显著高表达于系膜细胞增生性肾小球肾炎患儿肾活检组织中，且文献和实验结果均证实过表达AATF可显著增强系膜细胞的增殖能力。因此，我们有充分的理由提出科学假说：uc.412可能通过AATF调控系膜细胞的增殖活力。本课题拟阐述uc.412在儿童肾小球肾炎中的表达规律；通过过表达和沉默uc.412评判uc.412在系膜细胞增殖中的功能；以AATF为机制线索，探讨uc.412通过AATF调控系膜细胞增殖的分子机制。本研究将有助于阐明uc.412在肾小球系膜细胞增殖中的作用及机制，并可能为儿童MsPGN的防治提供新线索和潜在的干预靶标。

近年来，儿童系膜增生性肾小球肾炎(MsPGN)由于其临床表现多样、预后差，最终发展成慢性肾脏病(CKD)的趋势逐年升高。肾小球系膜细胞增生是导致肾小球硬化的主要原因之一，可以表现为系膜细胞的增殖及系膜区细胞外基质的过度沉积，但是具体机制并不清楚。LncRNA被证实在CKD的发生发展中发挥重要的调控作用。我们前期通过高通量lncRNA芯片筛选并进行验证发现lncRNA uc.412在异常增生的系膜细胞中显著上调，并通过GO和KEGG聚类分析发现uc.412可能参与调控细胞外基质、细胞增殖、细胞凋亡等过程。为此，我们进一步探讨uc.412在系膜细胞纤维化中的作用。我们在CKD小鼠肾脏组织中发现uc.412表达显著升高，体外实验中也证实uc.412参与了TGF-β1/Smad的信号通路机制调节，利用慢病毒过表达uc.412发现uc.412可以促进系膜细胞外基质沉积。利用RNA pull down和RNA测序等技术发现uc.412与RNA结合蛋白ELAVL1结合发挥作用，通过挽救实验进一步证实uc.412能够通过结合ELAVL1促进肾小球硬化。因此，uc.412可能是治疗MsPGN的理想靶点，对uc.412的机制探讨为MsPGN的防治提供了理论依据。