Department of Orthopaedics, Heze Municipal Hospital, No. 2888 Caozhou Road, Heze, 274031, China.
Department of Comprehensive Medical, Heze Infectious Disease Hospital, No. 298 Juyang Road, Heze, 274029, China.
Mol Cell Biochem. 2020 Jan;463(1-2):91-100. doi: 10.1007/s11010-019-03632-6. Epub 2019 Oct 12.
Baicalin (BAI), a sort of flavonoid monomer, acquires from Scutellaria baicalensis Georgi, which was forcefully reported in diversified ailments due to the pleiotropic properties. But, the functions of BAI in osteoblast differentiation have not been addressed. The intentions of this study are to attest the influences of BAI in the differentiation of osteoblasts. MC3T3-E1 cells or rat primary osteoblasts were exposed to BAI, and then cell viability, ALP activity, mineralization process, and Runx2 and Ocn expression were appraised through implementing CCK-8, p-nitrophenyl phosphate (pNPP), Alizarin red staining, western blot, and RT-qPCR assays. The microRNA-217 (miR-217) expression was evaluated in MC3T3-E1 cells or rat primary osteoblasts after BAI disposition; meanwhile, the functions of miR-217 in BAI-administrated MC3T3-E1 cells were estimated after miR-217 inhibitor transfection. The impacts of BAI and miR-217 inhibition on Wnt/β-catenin and MEK/ERK pathways were probed to verify the involvements in BAI-regulated the differentiation of osteoblasts. BAI accelerated cell viability, osteoblast activity, and Runx2 and Ocn expression in MC3T3-E1 cells or rat primary osteoblasts, and the phenomena were mediated via activations of Wnt/β-catenin and MEK/ERK pathways. Elevation of miR-217 was observed in BAI-disposed MC3T3-E1 cells or rat primary osteoblasts, and miR-217 repression annulled the functions of BAI in MC3T3-E1 cell viability and differentiation. Additionally, the activations of Wnt/β-catenin and MEK/ERK pathways evoked by BAI were both restrained by repression of miR-217. These explorations uncovered that BAI augmented the differentiation of osteoblasts via activations of Wnt/β-catenin and MEK/ERK pathways by ascending miR-217 expression.
黄芩苷(BAI)是一种黄酮类单体,从黄芩中提取,由于其多效性而在多种疾病中得到广泛报道。但是,BAI 在成骨细胞分化中的作用尚未得到解决。本研究旨在证实 BAI 对成骨细胞分化的影响。将 MC3T3-E1 细胞或大鼠原代成骨细胞暴露于 BAI 中,然后通过 CCK-8、对硝基苯磷酸(pNPP)、茜素红染色、western blot 和 RT-qPCR 检测细胞活力、碱性磷酸酶(ALP)活性、矿化过程和成骨相关基因 Runx2 和 Ocn 的表达。在 BAI 处理后,评估 MC3T3-E1 细胞或大鼠原代成骨细胞中的 microRNA-217(miR-217)表达;同时,在转染 miR-217 抑制剂后,评估 miR-217 在 BAI 处理的 MC3T3-E1 细胞中的功能。通过探究 BAI 和 miR-217 抑制对 Wnt/β-catenin 和 MEK/ERK 通路的影响,验证它们在 BAI 调节成骨细胞分化中的作用。BAI 可加速 MC3T3-E1 细胞或大鼠原代成骨细胞的细胞活力、成骨细胞活性以及 Runx2 和 Ocn 的表达,这一现象是通过激活 Wnt/β-catenin 和 MEK/ERK 通路介导的。在 BAI 处理的 MC3T3-E1 细胞或大鼠原代成骨细胞中观察到 miR-217 的上调,而 miR-217 的抑制消除了 BAI 对 MC3T3-E1 细胞活力和分化的作用。此外,BAI 诱导的 Wnt/β-catenin 和 MEK/ERK 通路的激活均被 miR-217 的抑制所抑制。这些研究结果表明,BAI 通过上调 miR-217 的表达,激活 Wnt/β-catenin 和 MEK/ERK 通路,增强成骨细胞的分化。