Lai Manxiang, Chen Xia, Feng Juan, Ruan Zhiyan, Lin Jiwei
Department of Pharmacy, Guangdong Food and Drug Vocational College Guangzhou 510520, Guangdong, China.
Department of Nursing, Guangdong Food and Drug Vocational College Guangzhou 510520, Guangdong, China.
Am J Transl Res. 2024 Sep 15;16(9):4492-4503. doi: 10.62347/WMLI2601. eCollection 2024.
To investigate the role of Morinda officinalis polysaccharide (MOP) in the protein expression of the Wnt/β-catenin signaling cascade during the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and to elucidate the mechanisms by which MOP enhances osteogenic differentiation at the cellular level.
BMSCs were isolated and cultured using the whole bone marrow adherence method, followed by flow cytometry for the detection of BMSC marker antigens. Two groups were prepared: a low-dose MOP (L-MOP, 10 µg/mL) group and a high-dose MOP (H-MOP, 40 µg/mL) group. MTT assays and cell clone formation assays were performed to evaluate the effects of different MOP doses on BMSC proliferation. Alizarin red staining (ARS) and alkaline phosphatase (ALP) staining were conducted to assess the impact of varying MOP doses on nodule calcification and ALP activity in BMSCs. Additionally, western blot assays were carried out to determine the effects of different MOP concentrations on the expression levels of osteogenesis-related factors and Wnt/β-catenin pathway proteins in BMSCs.
Highly purified BMSCs were successfully extracted. Subsequent assays demonstrated that BMSCs exhibited enhanced proliferation at all MOP doses, particularly at the H-MOP dose, compared to the control group. Both L-MOP and H-MOP increased calcium content and ALP activity in BMSCs, as well as elevated the expression of osteogenic factors and Wnt/β-catenin pathway proteins compared to the blank control group. However, the addition of Dickkopf-1 (DKK1) significantly reduced BMSC proliferation and osteogenic differentiation compared to the H-MOP group.
MOP can enhance BMSC proliferation and osteogenic differentiation by activating the Wnt/β-catenin signaling pathway.
探讨巴戟天多糖（MOP）在骨髓间充质干细胞（BMSCs）成骨分化过程中对Wnt/β-连环蛋白信号级联蛋白表达的作用，并在细胞水平阐明MOP增强成骨分化的机制。
采用全骨髓贴壁法分离培养BMSCs，然后通过流式细胞术检测BMSC标记抗原。制备两组：低剂量MOP（L-MOP，10μg/mL）组和高剂量MOP（H-MOP，40μg/mL）组。进行MTT试验和细胞克隆形成试验，以评估不同剂量MOP对BMSCs增殖的影响。进行茜素红染色（ARS）和碱性磷酸酶（ALP）染色，以评估不同剂量MOP对BMSCs结节钙化和ALP活性的影响。此外，进行蛋白质免疫印迹试验，以确定不同MOP浓度对BMSCs中成骨相关因子和Wnt/β-连环蛋白通路蛋白表达水平的影响。
成功提取了高度纯化的BMSCs。随后的试验表明，与对照组相比，所有剂量MOP下BMSCs的增殖均增强，尤其是在H-MOP剂量下。与空白对照组相比，L-MOP和H-MOP均增加了BMSCs中的钙含量和ALP活性，同时提高了成骨因子和Wnt/β-连环蛋白通路蛋白的表达。然而，与H-MOP组相比，添加Dickkopf-1（DKK1）显著降低了BMSCs的增殖和成骨分化。
MOP可通过激活Wnt/β-连环蛋白信号通路增强BMSCs的增殖和成骨分化。
