目的 采用生物信息學(xué)方法識別絕經(jīng)后骨質(zhì)疏松癥中免疫反應(yīng)相關(guān)的基因,并探討其作用機制和篩選靶向中藥活性成分。方法 在基因表達(dá)數(shù)據(jù)庫(GEO)中檢索"postmenopausal osteoporosis",下載基因數(shù)據(jù)集GSE230665,通過R軟件中的"limma"數(shù)據(jù)包獲取差異表達(dá)基因,使用David工具對差異基因進(jìn)行基因本體(GO)功能富集分析并篩選出免疫反應(yīng)相關(guān)的關(guān)鍵基因,然后對關(guān)鍵基因進(jìn)行GO功能富集分析和京都基因與基因組百科全書(KEGG)通路富集分析。采用STRING平臺和Cystoscope軟件建立蛋白相互作用(PPI)網(wǎng)絡(luò),用MCODE和CytoNCA插件識別重要基因簇模塊和核心基因。在CTD數(shù)據(jù)庫搜索"postmenopausal osteoporosis"對核心基因進(jìn)一步篩選并搜索靶向核心基因的中藥活性成分,并在PubChem數(shù)據(jù)庫中搜索中藥活性成分的3D結(jié)構(gòu),Autodock 4軟件進(jìn)行分子對接,以及探索核心基因與絕經(jīng)后骨質(zhì)疏松癥的相互作用。結(jié)果 在絕經(jīng)后骨質(zhì)疏松癥中篩選出43個關(guān)鍵基因,GO富集分析顯示主要參與涉及正向調(diào)節(jié)炎癥反應(yīng)、白細(xì)胞介素(IL)-12的產(chǎn)生、腫瘤壞死因子的產(chǎn)生、細(xì)胞遷移和趨化作用等;KEGG通路主要涉及趨化因子信號通路、磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信號通路、腫瘤壞死因子(TNF)信號通路、核因子-κB(NF-κB)信號通路等。鑒定出9個核心基因依次為IL-10、B2M、IL-17A、CXCR4、TLR4、CCR5、NOTCH1、TLR2、SMAD3。通過驗證,在絕經(jīng)后骨質(zhì)疏松癥患者中核心基因的表達(dá)均上調(diào)。核心基因?qū)?yīng)的中藥活性成分為雌二醇、白藜蘆醇、槲皮素、姜黃素、雙酚A和木黃酮等,這些中藥活性成分可能通過抑制免疫反應(yīng)來修復(fù)絕經(jīng)后骨質(zhì)疏松癥。結(jié)論 利用生物信息學(xué)篩選出與免疫反應(yīng)相關(guān)的核心基因(IL-10、B2M、IL-17A、CXCR4、TLR4、CCR5、NOTCH1、TLR2、SMAD3)以及其受靶向調(diào)控的主要中藥活性成分(雌二醇、白藜蘆醇、槲皮素、姜黃素、雙酚A和木黃酮),為進(jìn)一步探索絕經(jīng)后骨質(zhì)疏松癥的分子發(fā)病機制以及新的治療靶點提供新的理論依據(jù)。

Objective To identify genes related to immune response in postmenopausal osteoporosis by bioinformatics, and to investigate the mechanism of their action and screen the active ingredients of targeted traditional Chinese medicines. Methods Search for "postmenopausal osteoporosis" in the gene expression database (GEO), download the gene dataset GSE230665, and obtain the differentially expressed genes through the "limma" packet in R software. David tool was used to perform GO functional enrichment analysis for differential genes and screen out key genes related to immune response, and then GO functional enrichment analysis and KEGG pathway enrichment analysis were performed for key genes. STRING platform and Cystoscope software were used to establish protein interaction (PPI) network, and MCODE and CytoNCA plug-ins were used to identify important gene cluster modules and core genes. Search for "postmenopausal osteoporosis" in CTD database to further screen core genes and search for TCM active ingredients targeting core genes, search for 3D structure of TCM active ingredients in PubChem database, and perform molecular docking with Autodock 4 software. And explore the interaction between core genes and postmenopausal osteoporosis. Results 43 Key genes were selected in postmenopausal osteoporosis, and GO enrichment analysis showed that they were mainly involved in the positive regulation of inflammatory response, the production of IL-12, tumor necrosis factor, cell migration and chemotaxis. KEGG pathway mainly involves chemokine signaling pathway, PI3K/Akt signaling pathway, TNF signaling pathway, NF-κB signaling pathway, etc. 9 Core genes were identified as IL-10, B2M, IL-17A, CXCR4, TLR4, CCR5, NOTCH1, TLR2, and SMAD3. It was verified that the expression of core genes was up-regulated in postmenopausal osteoporosis patients. Estradiol, resveratrol, quercetin, curcumin, bisphenol A, and lignans showed high interaction indices with the core genes, and the active ingredients of these herbal medicines may repair postmenopausal osteoporosis by suppressing the immune response. Conlusion Bioinformatics was utilized to screen the core genes associated with immune response (IL-10, B2M, IL-17A, CXCR4, TLR4, CCR5, NOTCH1, TLR2, SMAD3) as well as their main TCM active ingredients (estradiol, resveratrol, quercetin, curcumin, bisphenol A, and lignoflavone) subject to targeted regulation, which will provide a new theoretical basis for further exploring the molecular pathogenesis as well as new therapeutic targets of postmenopausal osteoporosis.

R285

國家中醫(yī)藥管理局青年岐黃學(xué)者支持項目（國中醫(yī)藥人教發(fā)[2020]7號）；黑龍江省重點研發(fā)計劃（GZ20210136）；黑龍江省中醫(yī)藥科研項目（ZHY2023-099）