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- 2018
miR-126修饰的间质干细胞来源的外泌体对大鼠早期缺血性股骨头坏死的影响
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Abstract:
摘要：目的 观察微小RNA-126(miR-126)修饰的间质干细胞来源的外泌体(MSC-126-Exos)对类固醇激素诱导的早期缺血性股骨头坏死(SANFH)的治疗作用，并探讨其潜在机制。方法 分别以80mg/L MSC-126-Exos和MSC-Exos处理人脐静脉内皮细胞(HUVECs)并设PBS处理组为对照，MTT、划痕实验以及成管实验分别观察各组细胞增殖、迁移和毛细血管网的形成，Western blot检测HUVECs中血管内皮生长因子A(VEGFA)的表达。构建大鼠SANFH模型，分别注射等量MSC-126-Exos、MSC-Exos和PBS，6周后，通过对股骨头区域组织进行CD31免疫荧光染色及其Micro-CT扫描，观察股骨头血管数量及骨质情况。结果 体外实验中，MSC-126-Exos组的HUVECs的增殖、迁移和成管较MSC-Exos组显著增强，且VEGFA的表达水平也上调，差异有统计学意义(P<0.05)。在体实验中，MSC-126-Exos组大鼠股骨头坏死区CD31免疫荧光标记的血管数量较MSC-Exos组和模型组显著增多(P<0.05)，且股骨头坏死区Micro-CT扫描显示MSC-126-Exos组的小梁厚度(Tb.Th)、小梁数(Tb.N)和每组织体积骨量(BV/TV)值也显著高于MSC-Exos组和模型组，而小梁分离度(Tb.Sp)显著降低(P<0.05)。结论 MSC-126-Exos可以促进内皮细胞功能，改善股骨头区的血运从而增强骨质沉积，对大鼠SANFH具有显著的治疗效果。
ABSTRACT: Objective To investigate the effects of exosomes derived from mesenchymal stem cells modified by miR-126 (MSC-126-Exos) on early steroid-induced avascular necrosis of the femoral head (SANFH) so as to explore its underlying mechanism. Methods The well-growing HUVECs were treated with 80mg/L of MSC-126-Exos, MSC-Exos and equal amount of PBS, respectively. The proliferation, migration and capillary network formation of HUVECs were detected by MTT, scratch test and tube formation assay, respectively. Western blot was used to detect the expression of vascular endothelial growth factor A (VEGFA) in HUVECs. Early steroid-induced avascular necrosis of femoral head (SANFH) models of Sprague Dawley (SD) rats were injected with MSC-126-Exos, MSC-Exos and PBS, respectively. After 6 weeks, CD31 immunofluorescence staining and Micro-CT scanning were performed on the region of the femoral head to evaluate the therapeutic effects of MSC-126-Exos. Results In vitro, MSC-126-Exos group significantly promoted the proliferation, migration and tube formation of HUVECs, and the expression level of VEGFA was also significantly higher than that in MSC-Exos group (P<0.05). In vivo, the number of CD31 immunofluorescence labeled vessels in the femoral head in MSC-126-Exos group was significantly larger than that in MSC-Exos group and model group (P<0.05). Besides, the Micro-CT scanning of the femoral head necrosis region showed that the trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), trabecular number (Tb.N) and bone volume per tissue volume (BV/TV) value in MSC-126-Exos group were significantly higher than those in MSC-Exos group and model group (P<0.05). Conclusion MSC-126-Exos can promote the function of endothelial cells, improve blood supply in the femoral head area and enhance the deposition of bone, thus having significant therapeutic effects on SANFH in rats
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