大鼠骨质疏松模型的建立及其骨髓间充质干细胞内差异lncRNA初步分析

Establishment of rat osteoporosis model and preliminary analysis of the differences of lncRNA in BMSC cells of model rats

目的:分析骨质疏松模型大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)内差异长链非编码RNA(long noncoding RNA,lncRNA)及相关功能。方法:SD大鼠分为骨质疏松模型组和对照组,每组6只,用切除卵巢的方法制备骨质疏松模型,通过双能X线骨密度检测仪测定两组大鼠手术前及3个月后的骨密度。取SD大鼠股骨内骨髓体外培养BMSC,利用基因芯片技术检测骨质疏松模型组(3只)和对照组(3只)大鼠BMSC内lncRNA的表达差异,通过GO功能分析及KEGG通路富集,分析差异lncRNA的生物学功能及相关信号通路。结果:骨质疏松模型组SD大鼠骨密度[(157.33±6.28)mg/cm^(2)],较对照组[(183.33±4.50)mg/cm^(2)]显著降低。两组大鼠BMSC内检测出基因片段共计32759个,差异有统计学意义的有8454个(P<0.05),其中上调3593个,下调4861个。检测到的lncRNA基因片段4992个,差异有统计学意义的共116个(P<0.05),其中上调35个,下调81个。差异lncRNA主要参与细胞生长发育生物学过程,并调控蛋白的转运通路。结论:骨质疏松模型大鼠的BMSC内lncRNA表达和正常大鼠存在差异,差异表达的lncRNA参与对BMSC生长发育过程及蛋白转运通路的调控。

Objective:To analyze the differences of lncRNA in cultured bone marrow mesenchymal stem cells(BMSCs)in osteoporotic model rats and the functions of the lncRNA.Methods:Twelve SD rats were equally divided into the osteoporosis model group and the control group.The osteoporosis model was made by ovariectomy.Bone mineral density(BMD)in ovariectomized rats and sham control were determined by dual-energy X-ray bone mineral density detector.BMSCs from femur bone marrow were cultured in vitro.The different expression of lncRNA in BMSCs of osteoporosis model(n=3)and control group(n=3)was detected by gene chip technology,and the different biological functions of lncRNA and related signaling pathway were analyzed by GO function analysis and the KEGG enrichment.Results:BMD in osteoporosis model rats[(157.33±6.28)mg/cm^(2)],was lower than the control group rats[(183.33±4.50)mg/cm^(2)]significantly(P<0.05).A total of 32759 gene fragments were detected in BMSCs in the osteoporosis model group and the control group,and 8454 lncRNA gene fragments had significant difference(P<0.05),among which 3593 were upregulated and 4861 were down-regulated in osteoporosis model group.Totally 4992 lncRNA gene fragments were detected,and there were significant differences in 116 lncRNA gene fragments in BMSCs of osteoporosis model rats,with 35 increased and 81 decreased.Differential lncRNAs are mainly involved in the biological process of cell growth and development,and regulate protein transport pathways.Conclusion:There were differences in the expression of lncRNA in BMSCs between osteoporosis model rats and normal rats,and the differentially expressed lncRNA involved in the regulation of BMSCs growth and development process or protein transport pathway.