杜仲促进去势大鼠牙槽骨成骨的作用  

Eucommia ulmoides promotes alveolar bone formation in ovariectomized rats

作　　者：郑琳 靳文君 罗珊珊 黄芮 王杰 程余婷 安哲庆 熊玥 巩仔鹏[2] 廖健 Zheng Lin;Jin Wenjun;Luo Shanshan;Huang Rui;Wang Jie;Cheng Yuting;An Zheqing;Xiong Yue;Gong Zipeng;Liao Jian(School/Hospital of Stomatology,Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Guizhou Provincial Engineering Research Center for Development and Application of Ethnic Medicines and Traditional Chinese Medicines,Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学口腔医学院/附属口腔医院,贵州省贵阳市550004 [2]贵州医科大学民族药与中药开发应用教育部工程研究中心,贵州省贵阳市550004

出　　处：《中国组织工程研究》2025年第6期1159-1167,共9页Chinese Journal of Tissue Engineering Research

基　　金：贵州省基础研究计划重点项目(黔科合基础-ZK[2023]重点037),项目负责人:廖健;贵州省卫生健康委科学技术基金项目(gzwkj2023-439),项目负责人:熊玥。

摘　　要：背景:杜仲具有一定的成骨作用,能够促进成骨细胞的增殖及分化,但其对牙槽骨骨形成及Wnt/β-连环蛋白信号通路的影响尚不明确。目的:基于Wnt/β-连环蛋白信号通路探讨杜仲促进去势大鼠牙槽骨成骨的作用机制。方法:采用随机数字表法将60只雌性SD大鼠分为空白对照组、假手术组、模型组、杜仲低剂量组、杜仲高剂量组,每组12只。模型组、杜仲低剂量组、杜仲高剂量组通过双侧卵巢切除术建立骨质疏松大鼠模型,假手术组切除双侧卵巢周围同等质量的脂肪组织。造模3个月后,杜仲低、高剂量组大鼠分别灌胃给予2.1,4.2 g/(kg•d)的杜仲,假手术组和模型组大鼠灌胃给予生理盐水。药物干预12周后取材,通过Micro-CT观察牙槽骨骨量变化,苏木精-伊红染色观察牙槽骨病理结构变化,ELISA法检测血清中碱性磷酸酶、骨钙素水平,Western blot检测牙槽骨组织中β-连环蛋白、卷曲蛋白9的蛋白表达,RT-qPCR检测牙槽骨组织中骨钙素、Runx2、碱性磷酸酶、β-连环蛋白、卷曲蛋白9 mRNA的表达。结果与结论:①与空白对照组比较,模型组骨体积分数、骨小梁数目、骨小梁厚度、骨密度均降低(P<0.05),骨小梁分离度升高(P<0.05);病理观察可见骨小梁排列紊乱、不规则,骨小梁变细或断裂,骨髓腔变大,骨量明显减少;血清中碱性磷酸酶水平上升(P<0.05),骨钙素水平下降(P<0.05);碱性磷酸酶、骨钙素、Runx2、β-连环蛋白、卷曲蛋白9的mRNA表达均降低(P<0.05);β-连环蛋白、卷曲蛋白9的蛋白表达降低(P<0.05)。②与模型组比较,杜仲低、高剂量组大鼠牙槽骨骨体积分数、骨小梁数目、骨小梁厚度、骨密度均升高(P<0.05),骨小梁分离度降低(P<0.05);杜仲低、高剂量组骨小梁排列稍整齐,骨小梁较粗大,骨量增加;杜仲低、高剂量组血清中碱性磷酸酶水平下降(P<0.05),骨钙素水平上升(P<0.05);杜仲低剂量组碱性磷酸酶�BACKGROUND:Eucommia ulmoides has a certain osteogenic effect,which can promote the proliferation and differentiation of osteoblasts.However,it is unclear whether Eucommia ulmoides has effects on alveolar bone formation and Wnt/β-Catenin signaling pathway.OBJECTIVE:To investigate the mechanism by which Eucommia ulmoides promotes alveolar bone formation in ovariectomized rats based on the Wnt/β-Catenin signaling pathway.METHODS:Sixty female Sprague-Dawley rats were selected and randomly divided into five groups:blank control group,sham-operation group,model group,low-dose group Eucommia ulmoides group,and high-dose Eucommia ulmoides group,with twelve rats in each group.Osteoporosis animal models were constructed by bilateral oophorectomy in the model group and the low-dose and high-dose Eucommia ulmoides groups.The sham-operation group underwent the same method to remove adipose tissue of equal mass around the bilateral ovaries.Three months after surgery,the low-and high-dose Eucommia ulmoides groups were given 2.1 g/kg/d and 4.2 g/kg/d Eucommia ulmoides by gavage,respectively.The sham-operation group and model group were given the same amount of physiological saline by gavage.After 12 weeks of drug intervention,the changes in alveolar bone mass of rats in each group were observed through Micro-CT;hematoxylin-eosin staining was used to observe the pathological structural changes of alveolar bone in rats;enzyme linked immunosorbent assay was used to detect the expression levels of alkaline phosphatase and osteocalcin in the serum of rats;western blot was used to detect the expression levels ofβ-Catenin and Frizzled9 receptor proteins in the alveolar bone of rats;and real-time fluorescence quantitative PCR was used to detect the expression of osteocalcin,Runt-related transcription factor 2(Runx2),alkaline phosphatase,β-catenin,and frizzled9 mRNAs in alveolar bone tissues of rats.RESULTS AND CONCLUSION:Compared with the blank control group,bone volume fraction,trabecular number,trabecular thickness,and bone minera

关 键 词：杜仲 牙槽骨 骨形成 去势大鼠 Wnt/β-连环蛋白信号通路 骨质疏松 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R285.6