王军 王剑 鲁敏 牧亭亭 耿亚会

摘要 目的：探討异补骨脂素通过PPAR-γ-Axin2-Wnt信号通路调节对大鼠骨代谢的影响。方法：选择雌性大鼠40只，随机分为模型组和异补骨脂素组，各20只，实验12周后，对大鼠骨密度、骨组织形态进行观察，并检测各组大鼠血清骨代谢生化指标情况，采用蛋白质免疫印迹法（Western blotting）检测PPAR-γ、Axin2及β-catenin蛋白的表达情况。结果：12周后，模型组大鼠股骨、骨盆、脊柱及全身骨密度值均低于异补骨脂素组（P<0.05）;异补骨脂素组大鼠血清钙、磷及抗酒石酸酸性磷酸酶（Tartrate Resistant Acid Phosphatase，TRACP）水平低于模型组，血清碱性磷酸酶（Alkaline Phosphatase，ALP）水平高于模型组（P<0.05）;骨组织形态计量学分析结果显示，模型组大鼠骨小梁稀疏、断裂明显，而异补骨脂素组大鼠骨小梁面积百分数和骨小梁数目明显增加，骨小梁分离度明显下降（P<0.05）。12周后，与模型组比较，异补骨脂素组PPAR-γ和Axin2蛋白水平下调，β-catenin蛋白表达明显增加（P<0.05）。结论：异补骨脂素可能通过抑制Axin2/PPAR-γ信号通路，激活Wnt信号通路，改善大鼠骨代谢。

关键词 异补骨脂素;骨质疏松;骨代谢;骨组织形态;Wnt信号通路;PPAR-γ蛋白;Axin2蛋白;β-catenin蛋白

Effects of Isopsoralen on Bone Metabolism in Rats Via PPAR Gamma-Axin 2-Wnt Signal Regulation

WANG Jun，WANG Jian，LU Min，MU Tingting，GENG Yahui

（Department of Traumatic Orthopaedics，People′s Hospital of Inner Mongolia Autonomous Region，Hohhot 010000，China）

Abstract Objective：To investigate the effects of isopsoralen on bone metabolism in rats through PPAR-gamma-Axin2-Wnt signal regulation.Methods：A total of 40 female rats were randomly divided into a model group and an isopsoralen group with 20 rats in each group.After 12 weeks of experiment，the bone mineral density（BMD） and bone histomorphology of rats were observed，and the biochemical indexes of bone metabolism in serum of rats in each group were detected.The expressions of PPAR-gamma，Axin 2 and beta-catenin protein were detected by Western blotting.Results：After 12 weeks，BMD values of femur，pelvis，spine and whole body in model group were lower than those in isopsoralen group（P<0.05）; serum Ca，P and TRACP levels in isopsoralen group were lower than those in model group，and serum ALP levels were higher than those in model group（P<0.05）; bone histomorphometric analysis showed that the trabeculae of model group were sparse and fractured obviously.The percentage of bone trabecular area and the number of bone trabeculae increased significantly，and the degree of trabecular segregation decreased significantly（P<0.05）.After 12 weeks，compared with model rats，PPAR-gamma and Axin 2 protein levels in isopsoralen group were down-regulated and beta-catenin protein expression was significantly increased（P<0.05）.Conclusion：Isopsoralen may improve the abnormal bone metabolism in rats by inhibiting Axin2/PPAR gamma signaling pathway，activating Wnt signaling pathway and improving bone metabolism in rats.

Keywords Isopsoralen; Osteoporosis; Bone metabolism; Bone histomorphology; Wnt signaling pathway; PPAR-γ protein; AXIN2 protein; β-Catenin protein

中图分类号：R284文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.09.011

骨质疏松（Osteoporosis，OP）是临床常见的全身性骨骼疾病，其特征表现为骨组织微结构受损、骨量降低及骨脆性增加[1]。OP易导致骨折，引起患者致残和致死率增高，严重影响患者的生命质量[2]。目前OP已成为全球范围的一个公共健康问题，因此，对OP的防治具有重要的临床意义[3]。目前临床上OP的药物治疗虽然具有一定疗效，但其不良反应较多，且经济成本较高，因此探寻一种安全、不良反应小、费用低廉的药物治疗方法显得十分必要[4]。异补骨脂素是从豆科植物中提取的一种香豆素类化合物，具有促进骨代谢的作用[5]。本研究通过动物实验，探讨异补骨脂素调控大鼠PPAR-γ-Axin2-Wnt信号通路的机制及对骨代谢的影响，为防治OP提供新思路。

1 材料与方法

1.1 材料

1.1.1 动物 选用6月龄雌性健康大鼠[动物批准号：SCXK（京）2018-0013]40只，均购于北京华阜康生物科技股份有限公司，饲养于清洁级实验室中，所有大鼠自由饮水摄食。本研究经我院实验动物管理与动物福利伦理委员会审批通过（伦理审批号：20180121）。……