罗汉果苷V通过Wnt/β-catenin信号通路对糖尿病状态成骨细胞增殖与分化的影响

目的:探索罗汉果苷V(mogroside V，MV)对糖尿病状态下成骨细胞增殖分化的影响及其可能的作用机制。方法:构建糖尿病大鼠模型，分离培养正常大鼠和糖尿病大鼠下颌骨成骨细胞。CCK8法检测不同质量浓度MV对糖尿病大鼠成骨细胞的毒性和增殖活性，并筛选出适宜的MV浓度用于后续实验;取正常大鼠和糖尿病大鼠下颌骨成骨细胞，分别设立正常对照组、高糖组及MV+高糖组，茜素红染色及定量分析检测成骨分化情况，实时荧光定量PCR检测成骨相关因子ALP、OCN及Wnt/β-catenin信号通路相关因子Runx2，β-catenin，Axin2，Lef1及Osterix的mRNA表达。结果:0~0.4 g·L^-1质量浓度范围内MV对糖尿病状态成骨细胞均未表现出毒性，且浓度为6.25×10^-3g·L^-1时促进细胞增殖活性最强;与高糖组相比，MV+高糖组(含质量浓度为6.25×10^-3g·L^-1 MV)成骨分化能力增强，ALP、OCN、Runx2、β-catenin、Lef1、Osterix的表达显著升高，Axin2的表达显著降低。 结论:罗汉果苷V可促进糖尿病状态成骨细胞增殖与分化，其机制可能与调控Wnt/β-catenin信号通路有关。

Effect of mogroside V on the proliferation and differentiation of osteoblasts under the diabetic condition through Wnt/β-catenin signaling pathway

OBJECTIVE To explore the effect of mogroside V(MV)on the proliferation and differentiation of osteoblasts under diabetic conditions and its potential mechanism.METHODS Diabetic rat models were established,and mandibular osteoblasts of normal rats and diabetic rats were isolated and cultured,CCK8 method was used to detect the toxicity and proliferation activity of different mass concentrations of MV on osteoblasts in diabetic rats,and to screen out the appropriate MV concentration for subsequent experiments.The normal control group,high glucose group and MV+high glucose group were established by taking mandibular osteoblasts from the normal rats and the diabetic rats.,respectively,and then alizarin red staining was used to detect cell osteogenic differentiation,and quantitative real-time PCR was used to detect the mRNA expression of bone formation related factors ALP,OCN and Wnt/β-catenin signaling pathway related factors Runx2,β-catenin,Axin2,Lef1 and Osterix.RESULTS MV in the mass concentration range of 0-0.4 g·L^-1 showed no toxicity to osteoblasts in diabetic state,and the mass concentration of 6.25×10^-3g·L^-1 had the strongest activity in promoting cell proliferation.Compared with high glucose group,MV+ high glucose group(contained MV with a mass concentration of 6.25×10^-3g·L^-1)exhibited greater bone differentiation,the expression of ALP,OCN,Runx2,β-catenin,Lef1 and Osterix were significantly increased,and the expression of Axin2 was significantly decreased..CONCLUSION Mogroside V can promote the proliferation and differentiation of osteoblasts under diabetic conditions,which may be related to the regulation of Wnt/β-catenin signaling pathway.