淫羊藿苷对大鼠股骨头坏死微血管内皮细胞的影响△

Effect of icariin on bone microvascular endothelial cells in femoral head necrosis models of rats

摘要:

[目的]进一步验证淫羊藿苷(icariin, ICA)是否可有效的调控激素诱导的股骨头 BMECs 中 miRNA-23b 表达失衡、 保护 BMECs 功能、预防激素性 ONFH 的发生。[方法] 90 只雌性 SD 大鼠随机三组，每组 30 只动物。模型组给予脂多糖大剂量甲强龙建立股骨头坏死模型，并给予生理盐水灌胃。ICA 组同模型组给药，并给予 ICA (每天 60 mg/kg) 灌胃，连续灌注 4 周。空白组仅给予等量生理盐水。4 周处理动物，取股骨头行组织学检查，另外，自股骨头分离股骨头微血管内皮细胞(bone microvascular endothelial cells, BMECs)进行鉴定、细胞凋亡检测和 miRNA-23b 表达的生物信息学分析。[结果]组织形态计量方面，模型组股骨头骨小梁的宽度和骨小梁面积明显低于空白组与 ICA 组(P＜0.05)，而空白组与 ICA 组之间无明显差异(P＞ 0.05)。模型组的空骨陷窝率明显高于 ICA 组和空白组 (P＜0.05)。细胞凋亡方面， BMECs 活细胞率由高至低依次为：空白组 ＞ ICA 组 ＞模型组(P＜0.05)，其中，模型组显著低于空白组和 ICA 组(P＜0.05)，而空白组和 ICA 组差异无统计学意义(P＞ 0.05)。早期凋亡细胞率由高至低依次为：模型组 ＞ICA 组 ＞空白组 (P＜0.05)，其中，模型组显著高于空白组 (P＜0.05)，而空白组与 ICA 组的差异无统计学意义 (P＞0.05)。生物信息分析表明：与空白组相比，模型组 miRNA-23b 表达下调 (P＜0.05)， ICA 组与空白组差异无统计学意义 (P＞0.05)。[结论] ICA 在此动物模型可有效预防激素性 ONFH 发生，同时可保护 BMECs， BMECs 中的 miRNA-23b 可能是 ICA 预防激素性 ONFH 的作用靶点。

Abstract:

[Objective] To further verify whether icariin (ICA) regulate the expression imbalance of mirNA-23B in hormone-induced femoral head bone microvascular endothelial cells (BMECs) , protect the function of BMECs, and prevent the occurrence of hormone-in- duced osteonecrosis of femoral head (ONFH) . [Methods] Ninety female SD rats were randomly assigned to three groups with 30 animals in each group. The animals in the model group were given high dose of methylprednisolone with lipopolysaccharide to establish femoral head necrosis model, and were gavaged with normal saline. Those in the ICA group were given the drugs as the model group, and were simultane- ously gavaged with ICA 60 mg/kg · d-1 intragastrically for 4 weeks. However, the rats in the blank group only received the same amount of normal saline. At 4 weeks, the femoral heads were harvested for histological examination. In addition, BMECs were isolated from the femo- ral heads for identification, detection of apoptosis and bioinformatics analysis of miRNA- 23B expression. [Results] In term of histomor- phometry, the width and area of trabecular bone in the model group were significantly lower than those in blank group and ICA group (P＜ 0.05) , while which was not significantl different between blank group and ICA group (P＞0.05) . The ratio of empty bone lacunae in model group was significantly higher than that in ICA group and blank group (P＜0.05) . In terms of cell apoptosis, the active cell ratios of BMECs were ranked from high to low: the blank group ＞ the ICA group ＞ the model group (P＜0.05) , which in the model group was significantly low- er than the blank group and ICA group (P＜0.05) , while there was no significant difference between blank group and ICA group (P＞0.05) . Conversely, the ratios of early apoptotic cells from high to low was: the model group ＞ the ICA group ＞ the blank group (P＜0.05) , in which the model group was significantly higher than the control group (P＜0.05) , whereas there was no a significant difference between blank group and ICA group (P＞0.05) . As results of bioinformation analysis, mirNA-23B expression was down-regulated significantly in the mod- el group compared with that in the blank group (P＜0.05) , but there was no a significant difference between the ICA group and the blank group (P＞0.05) . [Conclusion] The ICA does effectively prevent the occurrence of steroid-induced ONFH and protect BMECs in this ani- mal models, in which the mirNA-23B in BMECs may be the target of ICA to prevent steroid-induced ONFH.