Wnt/β-catenin信号通路相关蛋白在补肾活血颗粒含药血清影响成骨细胞过程中的表达及意义

作者	单位
许应星	浙江中医药大学第一临床医学院
肖鲁伟	浙江省中医骨伤研究所
吴岩	浙江中医药大学第一临床医学院
吴承亮	浙江省中医骨伤研究所
童培建	浙江省中医院骨科

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中文摘要:

目的观察Wnt/β-catenin信号通路相关蛋白在补肾活血颗粒含药血清影响成骨细胞过程中的作用,为补肾活血颗粒含药血清治疗骨质疏松症作用机制的阐明提供实验依据。方法体外分离培养乳鼠颅骨来源成骨细胞,分为空白血清对照组和补肾活血颗粒含药血清组。连续培养6天后,定量检测各组成骨细胞的碱性磷酸酶活性,并进行碱性磷酸酶染色;18天后进行茜素红染色;同时应用酶联免疫法检测各组成骨细胞中Wnt/β-catenin信号通路相关蛋白——β-连环蛋白(β-catenin)、低密度脂蛋白受体相关蛋白5(LRP5)和T细胞因子(TCF)的表达情况。结果补肾活血颗粒含药血清能显著增加成骨细胞碱性磷酸酶的表达和促进矿化结节的形成。同时补肾活血颗粒含药血清也显著上调β-catenin、LRP5和TCF的蛋白表达。结论补肾活血颗粒含药血清能显著增加成骨细胞的成骨活性和矿化能力,这种作用与Wnt/β-catenin信号通路密切相关。提示Wnt/β-catenin信号通路在补肾活血颗粒治疗骨质疏松症的过程中发挥着重要作用。

英文摘要:

Objective To investigate the effect of Wnt/β-catenin signal pathway protein in the effect of Bushen Huoxue Granule(BHG) containing serum on the osteoblast,and to provide necessary experimental reliance for its action of mechanism in treatment of osteoporosis.Methods The osteoblast from cranial bones of neonates rat were isolated and cultured in vitro,which was divided into the blank control group and the BHG containing serum group.The alkaline phosphatase(ALP) activities of the osteoblast in each group were quantitatively detected and the ALP staining was performed six days later.The alizarin red staining was performed eighteen days later.At the same time,levels of Wnt/β-catenin signal pathway protein—β-catenin,low density lipoprotein correlated protein 5(LRP 5),and T cell factor(TCF) of osteoblasts in each group were detected by ELISA.Results BHG containing serum could significantly increase the expression of ALP and promote the formation of mineralizing nodus in the osteoblasts.At the same time it also markedly up-regulated the expressions of β-catenin,LRP 5,and TCF in this process.Conclusion BHG containing serum could markedly increase ossify activities and mineralization of osteoblast.This action was closely correlated with Wnt/β-catenin signal pathway.So it indicated that Wnt/β-catenin signal pathway played a very important role in the treatment of the osteoporosis by BHG.

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