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首页> 中文期刊> 《中国组织工程研究》 >淫羊藿苷对激素诱导损伤骨微血管内皮细胞微小RNA表达的影响

淫羊藿苷对激素诱导损伤骨微血管内皮细胞微小RNA表达的影响

         
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BACKGROUND:MicroRNAs (miRNAs) are widely involved in regulation of physiological processes, such as human development, cel proliferation, differentiation, and apoptosis, angiogenesis and lipid metabolism. MiRNAs also play an important regulating role in the pathological process of femoral head necrosis. At present, the research about the effect of icarin on miRNA expression in glucocorticoid- induced avascular necrosis is stil in the exploratory stage, and the specific targets, possible regulation mechanism and signaling pathway remain unclear. OBJECTIVE:To explore the effect of icarin on miRNA expression of bone microvascular endothelial cels in steroids-induced human femoral head lesionsin vitro. METHODS: Bone microvascular endothelial cels in cancelous bone of the femoral head were isolated and harvested in vitro. Icarin preconditioning preceded establishment of models of glucocorticoid-induced bone microvascular endothelial cel injury. Differential expression profiles and transcriptomes in glucocorticoid and normal groups were tested by miRNA microarrays. The most differentialy expressed miR-23b and miR-339 in microarray analysis were further confirmed by real-time quantitative PCR, Meanwhile the effects of icarin on the expression of miR-23b-5p and miR-339-5p were detected. RESULTS AND CONCLUSION:According to the microarray analysis, one miRNA was up-regulated and four mi RNAs were down-regulated in the glucocorticoid group (fold > 2,P < 0.05). Results of RT-qPCR revealed that miR-23b was down-regulated and miR-339 up-regulated in the glucocorticoid group, which were in agreement with the microarray analysis (P < 0.05). Icarin pretreatment effectively prevented the imbalances of miR-23b expression induced by glucocorticoid (P < 0.01). These findings indicate that Icarin may participate in the pathological process of steroid-induced femoral head necrosis through regulating the expression of miR-23b.%背景：近年来研究证实微小RNA广泛参与调控人体发育、细胞增殖分化及凋亡、血管生成、脂质代谢等生理过程，同样微小RNA在股骨头坏死的病理过程中也具有重要的调控作用。作者所在实验室前期研究证实淫羊藿苷具有抑制激素诱导骨微血管内皮细胞凋亡的作用，并且可以显著提高骨微血管内皮细胞促血管生成蛋白粒细胞集落刺激因子的表达。但目前关于淫羊藿苷对骨微血管内皮细胞微小RNA的研究仍处于探索阶段，其具体作用靶点、可能的调节机制及信号通路尚不明确。　　目的：探索淫羊藿苷对激素诱导人股骨头骨微血管内皮细胞微小RNA表达的影响及其可能的机制。　　方法：采用作者所在实验室建立的方法分离、培养及鉴定人股骨头骨微血管内皮细胞，取第3代骨微血管内皮细胞，建立激素诱导损伤模型及淫羊藿苷预处理模型，采用微小RNA基因芯片对激素组和正常组进行差异性转录，运用实时定量PCR法对明显表达差异miR-339及miR-23b进行验证，同时探讨淫羊藿苷对miR-339及miR-23b表达的影响。　　结果与结论：①对微小 RNA 基因芯片结果进行差异性分析，与正常组相比较，激素组中差异倍数﹥2的微小RNA共有5个，其中1个表达上调，4个表达下调；②实时定量PCR检测结果显示：与正常组相比，激素组中miR-339及miR-23b出现明显变化，与基因芯片结果一致。与激素组相比，淫羊藿苷预处理可以显著提高 miR-23b 的表达；③结果表明：淫羊藿苷可以有效预防激素诱导骨微血管内皮细胞损伤引起的miR-23b表达失衡，推测淫羊藿苷可能通过介导骨微血管内皮细胞miR-23b的表达参与激素性股骨头坏死病理过程的调控。

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