肾髓同治方防治小鼠去卵巢绝经后骨质疏松症的机制

Mechanism of Shensui Tongzhi Formula on preventing and treating ovariectomized postmenopausal osteoporosis in mice

目的:研究肾髓同治方对小鼠去卵巢绝经后骨质疏松症的治疗作用及其潜在机制。方法:雌性C57BL/6J野生型小鼠18只,随机分为假手术组、模型组、肾髓同治组;间充质祖细胞β-catenin^(Gli1ER)敲除小鼠18只,随机分为阴性对照组、β-catenin^(Gli1ER)组和β-catenin^(Gli1ER)+肾髓同治组。双侧卵巢切除法造绝经后骨质疏松症模型。灌胃8周后,Micro-CT观察股骨微结构变化,ABH染色观察组织病理学变化,实时荧光定量PCR法检测β-catenin、PPAR-γ、Osterix mRNA表达水平。结果:C57BL/6J小鼠中,与模型组比较,肾髓同治组小鼠的骨密度、骨体积分数、骨小梁数量显著提高(P<0.05),骨小梁面积显著增大(P<0.05),脂滴面积和直径显著减小(P<0.05),骨小梁分离度显著降低(P<0.05);肾髓同治方干预后显著提高β-catenin、Osterix mRNA表达水平(P<0.05),降低PPAR-γmRNA表达水平(P<0.01)。β-catenin^(Gli1ER)敲除小鼠中,与阴性对照组比较,β-catenin^(Gli1ER)组的骨密度、骨体积分数、骨小梁数量显著降低(P<0.05),骨小梁面积显著减小(P<0.05),骨小梁分离度显著提高(P<0.01),脂滴面积和脂滴直径显著增大(P<0.05),β-catenin^(Gli1ER)组与β-catenin^(Gli1ER)+肾髓同治组的指标差异均无统计学意义。结论:肾髓同治方对小鼠去卵巢绝经后骨质疏松症有一定的防治作用,其主要机制与调控间充质祖细胞Wnt/β-catenin信号通路有关。

Objective:To study the therapeutic effect and its underlying mechanism of Shensui Tongzhi Formula on mouse ovariectomized postmenopausal osteoporosis model.Methods:Eighteen female C57 BL/6 J wild-type mice were randomly divided into sham operation group,model group,kidney-marrow treatment group,and 18β-catenin conditional knockout mice(mesenchymal progenitor cellβ-catenin knockout mice)were randomly divided into negative-control group,β-catenin^(Gli1ER) group,andβ-catenin^(Gli1ER)+kidney-marrow treatment group,osteoporosis model caused by bilateral ovariectomy.After 8 weeks,microcomputer tomography was used to observe femoral microstructure changes,histopathological examination,and Real-time fluorescence quantitative PCR to detectβ-catenin,PPAR-γ,and Osterix expression.Results:Compared with the C57 BL/6 J mouse model group,the bone mineral density,bone volume fraction,and bone trabecular of the kidney-marrow treatment group were significantly increased(P<0.05),the area of bone trabecular was significantly increased(P<0.05),however,the area and diameter of lipid droplets were significantly reduced(P<0.05),and the separation of trabecular bone decreased significantly(P<0.05).The intervention of Shensui Tongzhi Formula significantly increased the expression ofβ-catenin and Osterix mRNA(P<0.05),and decreased the expression of PPAR-γmRNA(P<0.01).Inβ-catenin gene knockout mice,compared with the negative control group,the bone density,bone volume fraction,and number of trabecular bone in theβ-catenin^(Gli1ER) group were significantly reduced(P<0.05),and the trabecular bone area was significantly reduced(P<0.05),the separation of trabecular bone was significantly increased(P<0.01),the area and diameter of lipid droplets were significantly increased(P<0.05).There were no significant index differences between theβ-catenin^(Gli1ER) group and theβ-catenin^(Gli1ER)+kidney-marrow treatment group.Conclusion:Shensui Tongzhi Formula has a certain preventive effect on postmenopausal osteoporosis in mice,whose mechanism is related to the regulation of Wnt/β-catenin signaling pathway in mesenchymal progenitor cells.