lncRNA 在绝经后骨质疏松症肾阴虚证中的表达特征及调控网络分析

摘要

Objective To investigate the lncRNA expression and regulation network in postmenopausal osteoporosis with kidney Yin deficiency.Methods The patients with postmenopausal osteoporosis were selected and randomly divided into: kidney Yin deficiency group (n=3) and kidney Yang deficiency group (n=3).Three healthy postmenopausal women were selected as control group.lncRNA gene expression in single nuclear cells in peripheral blood was examined using microarray method.The genes in kidney Yin deficiency group were compared to those in other 2 groups respectively.Common differential expressed genes were studied and the regulation network was established.Real-time RT-PCR was used to examine the expression of LINC00334, LINC00189, and LOC101929378 and the results of microarray were confirmed.Results Eight differential expressed lncRNAs, LINC00334 LINC00189, LOC101929378, XLOC_013921, ENSG00000237489.2, ENSG00000225278.2, AK125437, and RNA95672 were screened out from the comparison among the 3 groups.Pathway analysis indicated that 12 potential pathways corresponded to these aberrant lincRNAs.Real-time PCR confirmed that LINC00189 was down-regulated in the kidney Yin deficiency group (FC=4.71, P=0.007).LINC00334 (FC=6.83, P=0.005) and LOC101929378 (FC=4.51, P=0.035) were up-regulated in the kidney Yin deficiency group.The change trend was coincident with the result of CHIP.Conclusion Eight lncRNAs, LINC00334, LINC00189, LOC101929378, XLOC_013921, ENSG00000237489.2, ENSG00000225278.2, AK125437, and RNA95672, are involved in osteoporotic kidney Yin deficiency by regulation of Jak/Stat, MAPK, chemokine signaling pathway, and calcium signaling pathway in postmenopausal osteoporosis.%目的：探讨绝经后骨质疏松症肾阴虚证中lncRNAs的表达特征及基因调控网络。方法随机选择绝经后骨质疏松症受试者，分为肾阴虚证组3例和肾阳虚证组3例，并选择健康绝经后妇女3例为正常对照组，采用基因芯片技术检测外周血单个核细胞lncRNAs的表达水平。肾阴虚证组分别与其他2组比较，筛选共同差异表达lncRNAs，并构建lncRNA调控网络；实时荧光定量PCR检测LINC00334、LINC00189和LOC101929378的表达，验证基因芯片结果。结果肾阴虚证组与正常对照组和肾阳虚证组比较，筛选出8个共同差异表达 lncRNAs：LINC00334、LINC00189、LOC101929378、XLOC＿013921、ENSG00000237489.2、ENSG00000225278.2、AK125437和RNA95672；这些差异lncRNAs参与Jak／STAT、MAPK、胰岛素通路和钙离子代谢等12条信号转导通路的调控；实时荧光定量PCR证实，与正常对照组比较，LINC00189在绝经后骨质疏松症肾阴虚证表达下调（FC＝4.71，P＝0.007）；LINC00334（FC＝6.83，P＝0.005）和LOC101929378绝经后骨质疏松症肾阴虚证表达上调（FC＝4.51，P ＝0.035），变化趋势与芯片结果相一致。结论 LINC00334、LINC00189、LOC101929378、XLOC＿013921、AK125437、ENSG00000237489.2、ENSG00000225278.2和RNA95672等8条lncRNAs可能通过调控Jak／STAT、MAPK、胰岛素通路和钙离子代谢等信号通路参与绝经后骨质疏松症肾阴虚证的发生发展过程。