目的 基于差異表達(dá)基因分析與實(shí)驗(yàn)驗(yàn)證探討大黃素治療骨質(zhì)疏松癥的作用機(jī)制。方法 從基因表達(dá)綜合數(shù)據(jù)庫(kù)（GEO）中篩選骨質(zhì)疏松癥的芯片數(shù)據(jù)，經(jīng)GEO2R軟件處理獲取差異基因（DEGs），結(jié)合文獻(xiàn)分析選擇11β-羥基類固醇脫氫酶1（11β-HSD1）作為核心基因進(jìn)行研究；使用AutoDock軟件分子對(duì)接模擬大黃素與11β-HSD1的結(jié)合活性；小鼠骨髓間充質(zhì)干細(xì)胞（BMSCs）分別使用DMEM/F12培養(yǎng)基（對(duì)照組）、成骨分化專用培養(yǎng)基（成骨誘導(dǎo)組）、含不同濃度（2.5、5.0、10.0 μmol·L^-1）大黃素的成骨分化專用培養(yǎng)基（大黃素組）進(jìn)行培養(yǎng)，培養(yǎng)至14、21 d進(jìn)行堿性磷酸酶（ALP）和茜素紅染色觀察判斷成骨細(xì)胞分化成熟情況；培養(yǎng)至第7天時(shí)ELISA法檢測(cè)細(xì)胞中ALP水平，實(shí)時(shí)熒光定量PCR（qRT-PCR）檢測(cè)11β-HSD1與Runt相關(guān)轉(zhuǎn)錄因子2（Runx2） mRNA水平，Western blotting檢測(cè)骨鈣素（OCN）、骨橋蛋白（OPN）蛋白表達(dá)水平。結(jié)果 從GEO數(shù)據(jù)庫(kù)中獲得44個(gè)與脂肪/成骨細(xì)胞分化相關(guān)的DEGs，在脂肪細(xì)胞分化組中RASD1、HSD11B1（又稱11β-HSD1）、RGS2等31個(gè)基因表達(dá)量顯著上調(diào)； SHRM、EGR1、TNS3等13個(gè)基因表達(dá)量顯著下調(diào)。分子對(duì)接顯示大黃素與11β-HSD1具有較好的結(jié)合活性。ALP/茜素紅染色觀察發(fā)現(xiàn)10 μmol·L^-1的大黃素組成骨細(xì)胞分化較多，與對(duì)照組差異明顯。與成骨誘導(dǎo)組相比，大黃素可顯著下調(diào)11β-HSD1 mRNA水平（P＜0.01），顯著上調(diào)ALP水平、Runx2 mRNA水平（P＜0.01、0.001）。Western blotting檢測(cè)結(jié)果顯示，與成骨誘導(dǎo)組相比，大黃素組OCN蛋白表達(dá)顯著升高（P＜0.001），OPN蛋白表達(dá)呈升高趨勢(shì)。結(jié)論 大黃素體外誘導(dǎo)小鼠BMSCs成骨細(xì)胞分化，可能通過(guò)抑制11β-HSD1表達(dá)，增加Runx2表達(dá)發(fā)揮作用。

Objective To explore the mechanism of emodin in the treatment of osteoporosis based on differential expression gene analysis and experimental verification. Methods Chip data related to osteoporosis was screened from the Gene Expression Omnibus (GEO) database. The differentially expressed genes (DEGs) were obtained through GEO2R software processing. Based on literature analysis, 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) was selected as the core gene for research. Molecular docking simulation was conducted using AutoDock software to assess the binding activity of emodin with 11β-HSD1. Mouse bone marrow mesenchymal stem cells (BMSCs) were cultured in DMEM/F12 medium (control group), osteogenic differentiation-specific medium (osteogenic induction group), and osteogenic differentiation-specific medium containing different concentrations (2.5, 5.0, 10.0 μmol·L^-1) of emodin (emodin group). At 14 and 21 days of culture, alkaline phosphatase (ALP) and alizarin red staining were performed to observe and determine the differentiation and maturation of osteoblasts. On the 7th day of culture, the ALP level in cells was detected by Elisa, and the mRNA levels of 11β-HSD1 and Runt-related transcription factor 2 (Runx2) were detected by real-time fluorescence quantitative PCR (qRT-PCR). The protein expression levels of osteocalcin (OCN) and osteopontin (OPN) were detected by Western blotting. Results A total of 44 DEGs related to adipocyte/osteoblast differentiation were obtained from the GEO database. In the adipocyte differentiation group, the expression levels of 31 genes, including RASD1, HSD11B1 (also known as 11β-HSD1), and RGS2, were significantly upregulated; the expression levels of 13 genes, including SHRM, EGR1, and TNS3, were significantly downregulated. Molecular docking showed that emodin had good binding activity with 11β-HSD1. ALP/alizarin red staining revealed that the 10 μmol·L^-1 emodin group had more osteoblast differentiation, with significant differences from the control group. Compared with the osteogenic induction group, emodin significantly downregulated the mRNA level of 11β-HSD1 (P <0.01) and significantly upregulated the ALP level and Runx2 mRNA level (P <0.01, 0.001). Western blotting results showed that compared with the osteogenic induction group, the OCN protein expression in the emodin group was significantly increased (P <0.001), and the OPN protein expression showed an increasing trend. Conclusion Emoxanthine induces osteoblast differentiation of mouse BMSCs in vitro, which may exert its effect by inhibiting the expression of 11β-HSD1 and increasing the expression of Runx2.

R285.5

新疆維吾爾自治區(qū)自治區(qū)衛(wèi)生健康青年醫(yī)學(xué)科技人才專項(xiàng)（ WJWY-202332）；新疆醫(yī)科大學(xué)第六附屬醫(yī)院自然科學(xué)基金資助項(xiàng)目（ LFYKYZX2023-7）；烏魯木齊市中醫(yī)藥科技創(chuàng)新項(xiàng)目（ ZYYMS-38）