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山奈酚通过上调 microRNA-101 促进 MC3T3-E1 细胞的增殖、迁移和分化。

Kaempferol promotes proliferation, migration and differentiation of MC3T3-E1 cells via up-regulation of microRNA-101.

机构信息

a Department of Orthopaedics , China-Japan Union Hospital of Jilin University , Changchun , China.

b Department of Orthopaedics , Qingdao West Coast New Area Central Hospital , Qingdao , China.

出版信息

Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):1050-1056. doi: 10.1080/21691401.2019.1591428.

Abstract

Osteoblast has been found to exert indispensable functions during fracture healing. This study aimed to investigate the effects of kaempferol, a natural flavonoid compound, on osteoblast proliferation, migration and differentiation, as well as possible molecular mechanisms. qRT-PCR was performed to measure the expression level of microRNA-101 (miR-101). Cell viability and migration were respectively assessed using cell counting kit-8 (CCK-8) assay and two-chamber migration assay. Relative alkaline phosphatase (ALP) activity was evaluated using p-nitrophenyl phosphate (pNPP) as substrate. Western blotting was used to detect the protein expression levels of key molecules involved in cell proliferation, migration, differentiation, and Wnt/β-catenin pathway. Kaempferol treatment significantly promoted MC3T3-E1 cell proliferation, migration, and differentiation. Mechanistically, kaempferol notably enhanced the expression level of miR-101 in MC3T3-E1 cells. Knockdown of miR-101 obviously weakened the promoting effects of kaempferol on MC3T3-E1 cell proliferation, migration, and differentiation. In addition, kaempferol remarkably activated Wnt/β-catenin pathway in MC3T3-E1 cells via up-regulating miR-101. This research further confirmed the promoting effects of kaempferol on osteoblast proliferation, migration, and differentiation. The up-regulation of miR-101 and activation of Wnt/β-catenin pathway caused by kaempferol play critical roles in the promoting effects of kaempferol on osteoblast proliferation, migration, and differentiation.

摘要

成骨细胞在骨折愈合过程中发挥着不可或缺的作用。本研究旨在探讨天然类黄酮化合物山奈酚对成骨细胞增殖、迁移和分化的影响及其可能的分子机制。采用 qRT-PCR 检测微小 RNA-101(miR-101)的表达水平。分别采用细胞计数试剂盒-8(CCK-8)检测法和双室迁移检测法检测细胞活力和迁移。采用对硝基苯磷酸酯(pNPP)作为底物评估相对碱性磷酸酶(ALP)活性。采用 Western blot 检测参与细胞增殖、迁移、分化和 Wnt/β-catenin 通路的关键分子的蛋白表达水平。山奈酚处理显著促进 MC3T3-E1 细胞增殖、迁移和分化。机制上,山奈酚明显增强了 MC3T3-E1 细胞中 miR-101 的表达水平。miR-101 的敲低明显削弱了山奈酚对 MC3T3-E1 细胞增殖、迁移和分化的促进作用。此外,山奈酚通过上调 miR-101 显著激活了 MC3T3-E1 细胞中的 Wnt/β-catenin 通路。本研究进一步证实了山奈酚对成骨细胞增殖、迁移和分化的促进作用。山奈酚上调 miR-101 和激活 Wnt/β-catenin 通路在山奈酚促进成骨细胞增殖、迁移和分化的作用中起关键作用。

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