High water-soluble curcuminoids-rich extract regulates osteogenic differentiation of MC3T3-E1 cells: Involvement of Wnt/β-catenin and BMP signaling pathway.
High water-soluble curcuminoids-rich extract regulates osteogenic differentiation of MC3T3-E1 cells: Involvement of Wnt/β-catenin and BMP signaling pathway.
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DOI：
10.1016/j.chmed.2021.01.003
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发表时间：
2021-10
影响因子：
3.8
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本研究旨在评价使用聚乙烯吡咯烷酮K30的固体分散体形式（CRE-SD）的高水溶性富含姜黄素的提取物（CRE）对MC 3 T3-E1细胞的成骨诱导的作用。使用微波辅助萃取结合Diaion® HP-20柱色谱法预纯化CRE。通过细胞活力、碱性磷酸酶（ALP）活性和茜素红S活性测定，检测CRE-SD在MC 3 T3-E1细胞中的成骨细胞增殖和分化能力。通过真实的时间PCR和蛋白质印迹分析评估成骨细胞特异性基因的mRNA表达和下划线机制。CRE-SD 50 µg/mL增加碱性磷酸酶（ALP）活性，这是MC 3 T3-E1细胞和非成骨小鼠多能细胞系C3 H10 T1/2中成骨细胞的早期分化标志物，表明CRE-SD的作用不是细胞类型特异性的。茜素红S活性显示出显着量的钙沉积在细胞与CRE-SD处理。CRE-SD还以剂量依赖性方式上调有利于成骨细胞分化的转录因子的mRNA表达水平，包括BMP-2、Runx 2和胶原1a。Western blot分析显示，头蛋白减弱CRE-SD促进的BMP-2和Runx 2蛋白的表达。siRNA介导的Wnt/β-catenin信号通路的阻断也消除了CRE-SD的影响，表明Wnt/β-catenin依赖性活性。抑制不同的信号通路可消除CRE-SD对ALP活性的影响，证实CRE-SD通过Wnt/β-catenin和BMP信号通路诱导MC 3 T3-E1细胞向成骨细胞分化。这些结果共同表明，CRE-SD可能是一种潜在的治疗骨质疏松症的治疗剂。
The present study aimed to evaluate the effect of a high water-soluble curcuminoids-rich extract (CRE) in a solid dispersion form (CRE-SD) using polyvinylpyrrolidone K30 on osteogenic induction of MC3T3-E1 cells. CRE was pre-purified using a microwave assisted extraction couple with a Diaion® HP-20 column chromatography. The osteoblastic cell proliferation and differentiation potentials of CRE-SD in MC3T3-E1 cells were tested by cell viability, alkaline phosphatase (ALP) activity, and Alizarin red S activity assays. The mRNA expressions of osteoblast-specific genes and underline mechanisms were assessed by a real time PCR and western blot analysis. CRE-SD 50 µg/mL increased alkaline phosphatase (ALP) activity, an early differentiation marker of osteoblasts in both MC3T3-E1 cells and non-osteogenic mouse pluripotent cell line, C3H10T1/2, indicating the action of CRE-SD was not cell-type specific. Alizarin red S activity showed a significant amount of calcium deposition in cells treated with CRE-SD. CRE-SD also upregulated the mRNA expression levels of transcription factors that favor osteoblast differentiation including Bmp-2, Runx2 and Collagen 1a, in a dose dependent manner. Western blot analysis revealed that noggin attenuated CRE-SD-promoted expressions of Bmp-2 and Runx2 proteins. siRNA mediated blocking of Wnt/β-catenin signaling pathway also annulled the influence of CRE-SD, indicating Wnt/β-catenin dependent activity. Inhibition of the different signaling pathways abolished the influence of CRE-SD on ALP activity, confirming that CRE-SD induced MC3T3-E1 cells into osteoblasts through Wnt/β-catenin and BMP signaling pathway. These results collectively demonstrate that CRE-SD may be a potential therapeutic agent for the treatment of osteoporosis.