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目的检测草酸钠诱导的人肾小管上皮细胞结晶肾损伤模型中与骨桥蛋白(OPN)相关的长链非编码RNA(lncRNA)的差异表达,并探讨OPN相关lncRNA在结晶肾损伤发病中的作用。方法利用草酸钠(20 mmol/L)刺激人肾近曲小管上皮细胞HK-2建立草酸钠诱导的结晶肾损伤肾小管上皮细胞模型。运用RNA干扰技术对其转染OPN小干扰RNA构建OPN敲低细胞模型(OPN-siRNA组),对照组细胞转染阴性随机对照序列(NC-siRNA组)。采用Arraystar公司提供的人类全基因组lncRNA芯片(V4.0)检测两组结晶肾损伤肾小管上皮细胞模型中与OPN相关的lncRNA和mRNA的差异表达水平,并采用生物信息学方法分析lncRNA芯片结果。结果全基因组lncRNA芯片结果显示草酸钠诱导的结晶肾损伤肾小管上皮细胞模型中与OPN相关的差异表达lncRNA共583个,其中OPN-siRNA组表达上调354个、下调229个;差异表达mRNA共235个,上调139个,下调96个。生物信息学分析结果显示差异表达lncRNA参与结晶肾损伤中细胞生长、酶活性调节等多种生物学过程。结论草酸钠体外诱导的结晶肾损伤肾小管上皮细胞模型中存在OPN相关的差异表达的lncRNA。
Objective To detect the differential expression of long chain non-coding RNA (lncRNA) associated with osteopontin (OPN) in sodium oxalate-induced renal tubular epithelial cell injury model and to explore the role of OPN-related lncRNA in the pathogenesis of renal tubulointerstitial injury . Methods Human renal proximal tubule epithelial cells (HK-2) were stimulated with sodium oxalate (20 mmol / L) to establish a model of renal tubular epithelial cells induced by sodium oxalate. The OPN knockdown cell model (OPN-siRNA group) was constructed by transfected with OPN small interfering RNA using RNA interference technology. The control group cells were transfected with negative random control sequence (NC-siRNA group). Human lncRNA chip (V4.0) provided by Arraystar Company was used to detect the differential expressions of lncRNA and mRNA relative to OPN in two groups of renal tubular epithelial cell models with renal injury, and the results of lncRNA were analyzed by bioinformatics method. Results Whole genome lncRNA microarray results showed that 583 differentially expressed lncRNAs were associated with OPN in the renal tubular epithelial cell model induced by sodium oxalate, of which 354 were up-regulated and 229 were down-regulated in OPN-siRNA group; 235 mRNAs were differentially expressed A, up 139, down 96. Bioinformatics analysis showed that different expression of lncRNA involved in the crystallization of kidney injury in cell growth, enzyme activity regulation and other biological processes. CONCLUSIONS: In contrast, OPN-related differentially expressed lncRNAs are present in renal tubular epithelial cell models induced by sodium oxalate in vitro.