摘要
目的观察柚皮苷对骨髓间充质干细胞(MSCs)成骨分化能力的影响,并且确定促进MSCs成骨分化的最佳柚皮苷浓度。方法用不同浓度的柚皮苷培养MSCs后,QPCR检测骨形态发生蛋白(BMP-2)表达量,同时CCK-8法检测不同浓度柚皮苷对MSCs增殖能力影响,确定最佳柚皮苷浓度。将最佳浓度柚皮苷与成骨诱导液分别或共同培养MSCs,观察其对成骨信号轴中Smad1/5/8,Runx2(Runt-related transcription factor-2),成骨细胞特异性转录因子(Osterix) mRNA及蛋白表达的促进作用。结果柚皮苷对BMP-2mRNA表达具有明显促进作用(P<0.05),并且其促进作用与浓度梯度相关。较高浓度(浓度>1×10-3mol/L)的柚皮苷能够降低MSCs增殖能力,差异有统计学意义(P<0.05),而1×10-4mol/L及以下柚皮苷浓度对MSCs增殖无明显影响,差异无统计学意义(P>0.05)。确定1×10-4mol/L为最佳柚皮苷浓度。柚皮苷组的Runx2、Smad1/5/8及OsterixmRNA及蛋白表达水平显著高于空白组(P<0.05)。而单一成骨诱导液组对MSCs成骨促进作用较单一柚皮苷弱,差异有统计学意义(P<0.05),柚皮苷与成骨诱导液共培养MSCs后,表达水平较单一柚皮苷组升高,差异有统计学意义(P<0.05)。结论柚皮苷能够促进MSCs成骨分化,并且这一促进作用与成骨信号轴BMP-2/Smads/Runx2/Osterix表达密切相关。
Objective To observe the influence of naringin on bone marrow mesenchymal stem cells osteogenic differentiation,and figure out the optimum concentration of naringin in promoting MSCs osteogenic differentiation. Methods MSCs were cultured with naringin in series of concentrations.QPCR was used to detect the expression level of bone morphogenetic protein(BMP-2). To determine the optimum concentration of naringin on MSCs proliferation, CCK-8 assay was adopted. MSCs were cultured in osteogenic differentiation medium with or without optimized naringin, as well as naringin only, followed by studying the promotion of osteogenic signalling axis Smad 1/5/8, Runx2(Runt-related transcription factor-2). Osterix mRNA and protein expression. Results Naringin significantly increased the expression of BMP-2 mRNA(P<0.05), and it was dose dependent.MSCs proliferation could be significantly reduced by naringin in concentration higher than 1×103 mol/L(P<0.05), while there was no obvious inhibition in concentration lower than 1×10-4 mol/L(P>0.05). Therefore, following experiments adopted 1×10-4 mol/L as the optimum concentration of naringin. The expression levels of Runx2, smad 1/5/8 and Osterix mRNA/protein in naringin groups were significantly higher than those in control groups(P<0.05). Furthermore, osteogenesis of MSCs in naringinonly group was higher than osteogenic differentiation medium-only group(P<0.05), while the co-cultured of osteogenic differentiation medium and naringin was the highest(P<0.05). Conclusion Naringin can promote osteogenic differentiation of MSCs,and it was related with the expression level of BMP-2/Smads/Runx2/Osterix.
作者
宁宇
刘想忠
汪伟
许海甲
杨傲飞
蔡寒涛
李章华
邹季
NING Yu;LIU Xiangzhong;WANG Wei;XU Haijia;YANG Aofei;CAI Hantao;LI Zhanghua;ZOU Ji(Hubei University of Chinese Medicine,Wuhan 430065;Tongren Hospital ofWuhan University/Wuhan No.3 Hospital,Wuhan 430060;Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430061)
出处
《湖北中医药大学学报》
2019年第1期9-14,共6页
Journal of Hubei University of Chinese Medicine
基金
国家自然科学基金项目(项目编号:81472103
81071463)
湖北省自然科学基金重点项目(项目编号:2015CFA079)
武汉市科技局应用基础研究计划项目(项目编号:2015061701011626)
关键词
柚皮苷
骨髓间充质干细胞
成骨分化
naringin
mesenchymal stem cells
osteogenic differentiation
