六味地黄丸干预绝经后骨质疏松症患者基因表达谱分析

目的探讨中药六味地黄丸干预绝经后骨质疏松患者的分子机制并分析预测其靶标和药物作用通路。方法检索并下载GEO中与采用六味地黄丸干预绝经后骨质疏松症的微阵列基因表达数据集。通过获取的基因表达数据集,分析六味地黄丸干预前后绝经后骨质疏松症患者血液中差异表达基因变化,并将这部分差异表达基因与在绝经后骨质疏松症患者和正常绝经女性血液中差异表达的基因取交集,获得两对比组中共同差异表达的基因。通过基因本体论数据库(GO)对获得的共同差异表达基因分别从生物过程(BP)、分子功能(MF)和细胞组分(CC)三方面进行功能富集分析;利用京都基因和基因组百科全书数据库(KEGG)进行通路富集分析。利用生物通用交互数据集库(BioGRID)分析获得与共同差异表达基因产物互作的蛋白质,并构建蛋白质-蛋白质互作关系(PPI)网络,采用Cytoscape软件实现蛋白质-蛋白质互作关系网络的可视化。应用中医药数据库(BATMAN-TCM)对六味地黄丸主要中药成分的靶标基因进行预测。结果六味地黄丸干预后,得到1 350个差异表达基因,其中上调表达基因322个,下调表达基因1 027个。58个共同差异表达基因与六味地黄丸干预绝经后骨质疏松的分子机制相关,六味地黄丸干预后43个基因下调,15个基因上调。六味地黄丸主要中药成分的10个共同差异表达基因靶标为ESR1, FGFR2, MED1, PGR, PRKCB, PTGS1, PTGS2, TRIM24, VDR, WNT4。与六味地黄丸干预组差异表达基因对比分析,ATF2, FBXW7以及RDX基因在干预后呈现显著差异表达。结论 ATF2, FBXW7和RDX基因为参与六味地黄丸干预绝经后骨质疏松分子调控机制的关键基因。六味地黄丸主要中药成分的10个共同靶标基因中,ESR1, FGFR2, MED1, WNT4为六味地黄丸干预治疗绝经后骨质疏松等相关内分泌疾病的潜在调控基因。

Analysis of altered gene expression in postmenopausal-osteoporosis patients treated with Liuwei Dihuang pills

Objective It is to explore the molecular mechanism for the intervention of Chinese medicine Liuwei Dihuang(LWDH) pills on postmenopausal osteoporosis(PMOP) patients and to predict its target and pathway of action. Methods Based on the obtained gene expression data set, the changes of differentially expressed genes in the blood of postmenopausal osteoporosis patients before and after the intervention of Liuwei Dihuang Wan were analyzed, and the differentially expressed genes were compared with those in postmenopausal osteoporosis patients and normal menopausal women. The differentially expressed genes are intersected to obtain genes that are commonly differentially expressed in the two comparison groups. Functional enrichment analysis of the common differentially expressed genes obtained from the biological process(BP), molecular function(MF), and cell component(CC) through the gene ontology database(GO);Kyoto gene and genome encyclopedia database(KEGG) was used to perform pathway enrichment analysis. The bio-interactive data set database(BioGRID) was used to analyze and obtain the protein interacting with the common differentially expressed gene products, and a protein-protein interaction relationship(PPI) network was constructed. The protein-protein interaction relationship network was visualized using Cytoscape software. The Chinese medicine database(BATMAN-TCM) was used to predict the target genes of the main Chinese medicine components of Liuwei Dihuang pill. Results After the intervention of Liuwei Dihuang pill, 1 350 differentially expressed genes were obtained, of which 322 genes were up-regulated and 1 027 genes were down-regulated. 58 common differentially expressed genes were related to the molecular mechanism of Liuwei Dihuang Wan’s intervention for postmenopausal osteoporosis. After Liuwei Dihuang Wan’s intervention, 43 genes were down-regulated and 15 genes were up-regulated. The 10 common differentially expressed gene targets of the main Chinese herbal ingredients of Liuwei Dihuang Wan were ESR1, FGFR2, MED1, PGR, PRKCB, PTGS1, PTGS2, TRIM24, VDR, WNT4. Compared with Liuwei Dihuang pill intervention group, the differentially expressed genes of ATF2, FBXW7 and RDX genes showed significant differential expression after intervention. Conclusion ATF2, FBXW7 and RDX genes are the key genes involved in Liuwei Dihuang Wan’s intervention in the molecular regulation mechanism of postmenopausal osteoporosis. Among the 10 common target genes of the main Chinese medicine components of Liuwei Dihuang Wan, ESR1, FGFR2, MED1, WNT4 are potential regulatory genes for Liuwei Dihuang Wan to intervene in the treatment of postmenopausal osteoporosis and other related endocrine diseases.