艾灸对去卵巢骨质疏松症大鼠骨髓间充质干细胞Wnt/β-catenin信号通路的影响
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Effect of Moxibustion on the Wnt/β-catenin Signaling Pathway in Bone Marrow Mesenchymal Stem Cells in Rats with Ovariectomized Osteoporosis
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    目的 观察艾灸三阴交、关元穴对去卵巢骨质疏松症大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)中Wnt/β-catenin信号通路的影响,探索艾灸影响骨代谢的内在机制。方法 将60只6月龄SD大鼠随机分为正常组(20只)和去卵巢组(40只),采用去卵巢法复制大鼠骨质疏松症模型,将去卵巢组再随机分为模型组、雌二醇组和艾灸组,每组10只。采用MTT法检测大鼠BMMSCs活性,酶联免疫吸附法检测大鼠BMMSCs中骨钙素(bone glaprotein,BGP)含量,实时荧光定量PCR法测定Wnt3a、低密度脂蛋白受体相关蛋白-5(low density lipoprotein receptor associated protein-5,LRP-5)、LRP-6、Dkk1、β-连环蛋白(β-catenin)和T细胞因子(T-cell factor, TCF)mRNA相对表达水平,Western blot法测定Wnt3a、Dkk1、β-catenin、TCF蛋白的表达水平。结果 在体外培养第12、16天时,艾灸可显著提高骨质疏松症大鼠降低的BMMSCs活性。艾灸可显著升高骨质疏松症大鼠BMMSCs中降低的BGP含量,显著上调BMMSCs中降低的LRP-5、LRP-6 mRNA的表达水平,显著上调降低的Wnt3a、β-catenin和TCF mRNA及其蛋白的表达水平,显著下调升高的Dkk1 mRNA及其蛋白的表达水平,差异均有统计学意义(P<0.05)。结论 艾灸三阴交、关元穴增加去卵巢大鼠BMMSCs活性,提高BGP含量,其机制与干预Wnt/β-catenin信号通路有关。

    Abstract:

    Objective To investigate the effect of moxibustion at Sanyinjiao and Guanyuan points on the Wnt/β-catenin signaling pathway in bone marrow mesenchymal stem cells(BMMSCs) of rats with ovariectomized osteoporosis, as well as the internal mechanism of moxibustion in affecting bone metabolism. Methods A total of 60 Sprague-Dawley rats, aged 6 months, were randomly divided into normal group with 20 rats and ovariectomized group with 40 rats. Ovariectomization was performed to establish a rat model of osteoporosis, and the ovariectomized group was further divided into model group, estradiol group, and moxibustion group, with 10 rats in each group. MTT assay was used to measure the activity of BMMSCs; ELISA was used to measure the content of bone Gla protein (BGP) in BMMSCs; quantitative real-time PCR was used to measure the relative mRNA expression of Wnt3a, low-density lipoprotein receptor associated protein-5 (LRP-5), LRP-6, Dkk1, β-catenin, and T-cell factor (TCF), and Western blot was used to measure the protein expression of Wnt3a, Dkk1, β-catenin, and TCF. Results On days 12 and 16 of culture in vitro, moxibustion significantly increased the reduced activity of BMMSCs in rats with osteoporosis. Moxibustion significantly increased the reduced content of BGP, the reduced mRNA expression of LRP-5 and LRP-6, and the reduced mRNA and protein expression of Wnt3a, β-catenin, and TCF in BMMSCs, and it also significantly downregulated the increased mRNA and protein expression of Dkk1 in BMMSCs (P<0.05). Conclusion Moxibustion at Sanyinjiao and Guanyuan can increase the activity of BMMSCs and the content of BGP, possibly by regulating the Wnt/β-catenin signaling pathway.

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姚长风,刘自兵,唐 巍,张 燕,刘存斌,汪宗宝,李斯亮.艾灸对去卵巢骨质疏松症大鼠骨髓间充质干细胞Wnt/β-catenin信号通路的影响[J].安徽中医药大学学报,2020,39(2):54-58

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  • 在线发布日期: 2020-04-10