Berberine coated mannosylated liposomes curtail RANKL stimulated osteoclastogenesis through the modulation of GSK3β pathway via upregulating miR-23a.
Berberine coated mannosylated liposomes curtail RANKL stimulated osteoclastogenesis through the modulation of GSK3β pathway via upregulating miR-23a.
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DOI：
10.1016/j.intimp.2019.105703
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发表时间：
2019-09
影响因子：
5.6
通讯作者：
Sali Sujitha;M. Rasool
中科院分区：
文献类型：
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作者：
Sali Sujitha;M. Rasool
药物诱导的microRNA表现出重要的治疗方法;然而，在骨病疾病（包括骨质疏松症和类风湿性关节炎）的治疗中的这种进展仍然不清楚。因此，高剂量的非特异性药物递送增加了副作用的风险并降低了药物治疗功效。因此，本研究旨在通过改变miR-23 a表达来检查小檗碱包被的甘露糖基化脂质体（ML-BBR）对RANKL（100 ng/ml）刺激的骨髓源性单核细胞/巨噬细胞（Bcl-2）的治疗作用。使用共聚焦显微镜的初步研究显示，在RANKL刺激的BBR中，ML-BBR成功内化。ML-BBR处理通过抑制磷酸化谷胱甘肽合成酶激酶β（p-GSK 3 β）介导的NFATc 1活化，消除了BMM细胞中破骨细胞形成的增加。因此，ML-BBR还减弱了骨降解酶（TRAP、组织蛋白酶K和MMP-9）的表达，从而抑制破骨细胞的骨吸收活性。此外，ML-BBR在基因水平上诱导miR-23 a的表达水平，这反过来又减弱了GSK 3 β/p-GSK 3 β的表达，如通过印迹分析所证实的。使用荧光素酶报告基因测定证实miR-23 a对GSK 3 β磷酸化的进一步抑制。而LY 2090314（GSK 3 β抑制剂）则抑制GSK 3 β/p-GSK 3 β蛋白水平的表达。然而，LY 2090314处理诱导了miR-23 a的基础水平表达，这是由于ML-BBR在上调miR-23 a水平以抑制GSK-3β磷酸化方面具有影响性作用。总之，我们的研究结果证实，通过Bcl 4对ML-BBR的优先内化有效地调节了RANKL/p-GSK 3 β通路，并可能通过miR-23 a的转录后基因沉默来减少破骨细胞介导的骨侵蚀。
Drug-induced microRNAs manifest significant therapeutic approaches; however, such progress in the treatment of osteopathic disorders including osteoporosis and rheumatoid arthritis still remains obscure. Contrarily, non-specific drug delivery, at high doses, increases the risk of side effects and reduces drug therapeutic efficacy. Accordingly, the present study was designed to examine the therapeutic effect of berberine coated mannosylated liposomes (ML-BBR) on RANKL (100 ng/ml) stimulated bone marrow-derived monocytes/macrophages (BMMs) via altering miR-23a expression. Initial studies using confocal microscopy showed successful internalization of ML-BBR in RANKL stimulated BMMs. Treatment with ML-BBR abrogated the increased osteoclast formation in BMM cells via inhibiting phosphorylated glutathione synthase kinase beta (p-GSK3β) mediated NFATc1 activation. Consequently, ML-BBR also attenuated the expression of bone-degrading enzymes (TRAP, cathepsin K and MMP-9) thereby inhibiting the bone resorptive activity of osteoclasts. Moreover, ML-BBR induced the expression levels of miR-23a at the gene level, which in turn attenuated GSK3β/p-GSK3β expression as confirmed via blotting analysis. Further miR-23a inhibition of the GSK3β phosphorylation was confirmed using luciferase reporter assay. Comparatively, LY2090314 (GSK3β inhibitor) treatment inhibited the protein level expression of GSK3β/p-GSK3β. However, LY2090314 treatment induced a basal level expression of miR-23a owing to the suggestion that ML-BBR has an influential role in upregulating miR-23a level to inhibit GSK-3β phosphorylation. Cumulatively, our findings endorsed that preferential internalization of ML-BBR by BMMs effectively modulated the RANKL/p-GSK3β pathway and curtailed the osteoclast-mediated bone erosion possibly through post-transcriptional gene silencing via miR-23a.