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

Effects of icariin on micro RNAs expression in bone microvascular endothelial cells in steroids-induced femoral head lesions

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mi R-339及mi R-23b进行验证,同时探讨淫羊藿苷对mi R-339及mi R-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mi R-339及mi R-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mi R-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mi R-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mi R-23b的表达参与激素性股骨头坏死病理过程的调控。

BACKGROUND： Micro RNAs（mi RNAs） are widely involved in regulation of physiological processes, such as human development, cell proliferation, differentiation, and apoptosis, angiogenesis and lipid metabolism. Mi RNAs also play an important regulating role in the pathological process of femoral head necrosis. At present, the research about the effect of icariin on mi RNA expression in glucocorticoidinduced avascular necrosis is still in the exploratory stage, and the specific targets, possible regulation mechanism and signaling pathway remain unclear. OBJECTIVE： To explore the effect of icariin on mi RNA expression of bone microvascular endothelial cells in steroids-induced human femoral head lesions in vitro. METHODS： Bone microvascular endothelial cells in cancellous bone of the femoral head were isolated and harvested in vitro. Icariin preconditioning preceded establishment of models of glucocorticoid-induced bone microvascular endothelial cell injury. Differential expression profiles and transcriptomes in glucocorticoid and normal groups were tested by mi RNA microarrays. The most differentially expressed mi R-23 b and mi R-339 in microarray analysis were further confirmed by real-time quantitative PCR, Meanwhile the effects of icariin on the expression of mi R-23b-5p and mi R-339-5p were detected. RESULTS AND CONCLUSION： According to the microarray analysis, one mi RNA was up-regulated and four mi RNAs were down-regulated in the glucocorticoid group（fold〉 2, P〈0.05）. Results of RT-q PCR revealed that mi R-23 b was down-regulated and mi R-339 up-regulated in the glucocorticoid group, which were in agreement with the microarray analysis（P〈0.05）. Icariin pretreatment effectively prevented the imbalances of mi R-23 b expression induced by glucocorticoid（P〈0.01）. These findings indicate that Icariin may participate in the pathological process of steroid-induced femoral head necrosis through regulating the expression of mi R-23 b.