基于蛋白组学探讨糖皮质激素性骨质疏松发病机制

目的:基于Label-free非标全蛋白组学方法探讨糖皮质激素性骨质疏松(glucocorticoid-induced osteoporosis,GIOP)的发病机制。方法:将12只SPF级SD雌性大鼠随机分为空白对照组和GIOP组。适应性饲养1周后,GIOP组按2.5 mg/kg体重于左右大腿内侧交替注射地塞米松,空白对照组注射等量的生理盐水,每周2次。造模8周末,取各组大鼠的胫骨进行病理切片,以确定造模成功。每组选取3个样本进行蛋白质组学检测,在质控后通过蛋白质的定性定量分析筛选差异蛋白,并通过生物信息学手段针对差异蛋白进行聚类分析,利用基因本体论(gene ontology,GO)、KEGG(Kyoto encyclopedia of genes and genomes,KEGG)等功能富集分析手段,以阐明糖皮质激素(glucocorticoids,GCs)诱发的GIOP的潜在病理机制。结果:与空白对照组比较.GIOP组大鼠骨小梁排列紊乱,分布不均且伴有断裂现象,骨空腔间隙增加,具有显著差异,证明造模成功。蛋白组学检测共鉴定47个差异表达蛋白,其中20个蛋白上调,27个蛋白下调。...

Pathogenesis of glucocorticoid-induced osteoporosis based on label-free mass proteomics

Objective To explore pathogenesis of glucocortocoid-induced osteoporosis(GIOP) based on label-free mass proteomics.Methods Twevle female Sprague-Dawley(SD) rats were randomly divided into two groups,named as sham group and GIOP group. After one-week adaptive feeding,the rats of GIOP group were administered with dexamethasone via intramuscular injection according to 2.5 mg/kg weighting,while the rats of sham group were administered with the same amount of saline,twice a week. The tibias of each group were collected after 8-week modeling and made pathological sections to confirm the success of modeling. Three samples of each group were picked up to perform label-free mass proteomics. After quality control,differentially expressed proteins were identified according to qualitative and quantitative analyses. Then gene ontology(GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis,cluster analysis as well as protein-protein interaction analysis were performed using bioinformatics analysis.Results Compared with sham group,the structure of bone trabecular in GIOP group showed abnormal arrangement,uneven distribution and obvious fragmentation,which could demonstrate successful modeling. A total of 47 differentially expressed proteins (DEPs) were identified including 20 up-regulated and 27 down-regulated proteins. The expression of protein nucleophosmin 1(NPM1),adipocyte plasma membrane associated protein (APMAP),cytochromec oxidase subunit 6A1 (COX6A1) and tartrate-resistant acid phosphatase (ACP5) showed a significant difference between two groups. KEGG results showed DEPs were enriched on metabolism-related pathways,immune-related pathways and AMP-activated kinase (AMPK) signaling pathway.Conclusion Protein NPM1,APMAP,COX6A1 and ACP5 showed a close relationship with pathogenesis of GIOP,which could serve as potential biomarkers of GIOP. AMPK signaling pathway played an important role in the occurrence and development of GIOP,which could be regarded as potential signaling pathway to treatment GIOP.