骨髓间充质干细胞外泌体miR-10a-5p下调紫外线抵抗相关基因(UVRAG)的表达调控系统性红斑狼疮

Bone marrow mesenchymal stem cell exosomes miR-10a-5p regulates systemic lupus erythematosus cells by down-regulating expression of UVRAG

目的:探讨BMMSCs移植对SLE外周血单个核细胞增殖和凋亡的调控机制,为SLE的靶向治疗提供理论支持。方法:超速离心分离BMMSCs来源外泌体。CCK-8检测SLE外周血单个核细胞增殖活性,流式细胞术检测细胞凋亡情况。qRT-PCR检测外周血单个核细胞和外泌体中miRNA水平。双荧光素酶报告基因验证miR-10a-5p与紫外线抵抗相关基因(UVRAG)靶向关系。Western blot检测外泌体表面标记物和外周血单个核细胞中UVRAG的表达水平。结果:成功分离BMMSCs来源外泌体,并发现其抑制SLE外周血单个核细胞增殖,促进其凋亡。qRT-PCR检测结果表明BMMSCs来源外泌体可显著上调SLE外周血单个核细胞中miR-10a-5p表达水平。双荧光素酶报告基因检测以及Western blot检测结果表明miR-10a-5p靶向下调UVRAG表达水平。结论:BMMSCs来源外泌体miR-10a-5p通过靶向下调UVRAG水平,抑制SLE外周血单个核细胞增殖并促进细胞凋亡。

Objective:To investigate the effect of bone marrow mesenchymal stem cells transplantation on proliferation and apoptosis of SLE peripheral blood mononuclear cell in systemic lupus erythematosus,and to provide theoretical support for the targeted therapy of systemic lupus erythematosus.Methods:Exosomes were isolated from bone marrow mesenchymal stem cells conditioned medium by ultracentrifugation.CCK-8 was used to detect the proliferation of SLE peripheral blood mononuclear cells(PBMCs),and apoptosis was detected by flow cytometry.qRT-PCR was used to detect miRNA levels in PBMCs and exosomes.The dual luciferase reporter gene verified the targeting relationship between miR-10a-5p and UVRAG.Western blot was used to detect the expression of the markers of exosome and UVRAG in PBMCs.Results:The BMMSCs-derived exosomes were successfully isolated and found to inhibit the proliferation of SLE BMCs and promote apoptosis.The results of qRT-PCR showed that BMMSCs-derived exosomes up-regulated the level of miR-10a-5p peripheral blood mononuclear cells in SLE.Dual luciferase reporter gene assay and Western blot assay showed that miR-10a-5p targeted down-regulated UVRAG expression.Conclusion:BMMSCs-derived exosomes miR-10a-5p down-regulate UVRAG,thereby inhibiting proliferation of PBMCs and promoting apoptosis.