中国骨质疏松杂志

萸蓉壮骨膏调控EZH2/Wnt3a/β-catenin通路促进绝经后骨质疏松大鼠成骨分化

Yurong Zhuanggu Gao promotes osteogenic differentiation of postmenopausal osteoporosis rats by regulating EZH2/Wnt3a/β-catenin pathway

DOI：10.3969/j.issn.1006-7108.2024.06.007

中文关键词: 萸蓉壮骨膏绝经后骨质疏松症 EZH2/Wnt3a/β-catenin通路成骨分化

英文关键词:Yurong Zhuanggu Gao postmenopausal osteoporosis EZH2/Wnt3a/β-catenin channel osteogenic differentiation

基金项目:甘肃省科技重点研发计划项目（23YFFA0068）；甘肃省教育厅高等学校科研项目（2023A-080）；甘肃省研究生创新之星项目（2023CXZX-760）

作者	单位
邢尚曼 1，2 郭超1，2 宋冰1，2 白敏1，2 汪湛东1，2 汪可欣1，2 徐晓艳3 张延英1，2 汪永锋1，2*	1.甘肃中医药大学，甘肃兰州 730000 2.甘肃省实验动物行业技术中心，甘肃兰州 730000 3.甘肃中医药大学附属医院，甘肃兰州 730000

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中文摘要:

目的基于EZH2/Wnt3a/β-catenin通路调控成骨分化探讨萸蓉壮骨膏对绝经后骨质疏松大鼠的治疗作用及机制。方法取SD大鼠复制绝经后骨质疏松模型，模型构建成功后随机分为模型组、阿仑膦酸钠片组[6.3 mg/(kg?周)]及萸蓉壮骨膏高[10.8 g/(kg?d)]、中[5.4 g/(kg?d)]、低剂量[2.7 g/(kg?d)]组，另选同龄大鼠作为假手术组。Micro CT 检测骨微结构，HE染色观察股骨组织病理形态；IF法检测大鼠股骨组织中成骨相关蛋白（Runx2、OSX、OPG）表达水平。RT-qPCR 及 Western blot法检测大鼠胫骨组织中EZH2、Wnt3a、β-catenin mRNA和蛋白表达水平。结果与假手术组比较，模型组大鼠骨微结构（BMD、BV/TV、Tb.Th、Tb.N和Tb.Sp）显著破坏（P＜0.05），骨小梁排列稀疏松散，结构出现不连续间断，脂滴累积较多，骨组织中成骨蛋白表达显著降低（P＜0.05），EZH2 mRNA和蛋白表达水平显著升高，Wnt3a、β-catenin mRNA和蛋白表达水平显著降低（P＜0.05）；与模型组比较，各给药组大鼠骨微结构显著改善（P＜0.05），各给药组不同程度改善模型大鼠骨组织病理结构，各给药组大鼠骨组织中成骨蛋白表达不同程度增加（P＜0.05），各给药组大鼠骨组织中EZH2 mRNA和蛋白表达水平显著降低，Wnt3a、β-catenin mRNA和蛋白表达水平显著增加（P＜0.05）。结论萸蓉壮骨膏能够显著改善绝经后骨质疏松大鼠骨流失，其作用机制与萸蓉壮骨膏调控EZH2/Wnt3a/β-catenin 通路促进成骨分化有关。

英文摘要:

Objective To investigate the therapeutic effect and mechanism of Yu Rong Zhuanggu Gao on postmenopausal osteoporosis rats based on EZH2/Wnt3a/β-catenin pathway regulating osteogenic differentiation. Methods The postmenopausal osteoporosis model of SD rats was selected and randomly divided into model group, Alendronate sodium tablet group (6.3 mg/kg/w), Yu Rong Zhuangu Gao high (10.8g/kg/d), medium (5.4g/kg/d) and low dose (2.7g/kg/d) groups after the model was successful, and other rats of the same age were selected as sham operation group. The bone microstructure was detected by Micro CT and the histopathologic morphology of femur was observed by HE staining. The expression levels of osteogenic proteins (Runx2, OSX, OPG) in rat femur were detected by IF method. The mRNA and protein expression levels of EZH2, Wnt3a and β-catenin were detected by RT-qPCR and Western Blot. Results Compared with sham operation group, bone microstructure (BMD, BV/TV, Tb.Th, Tb.N and Tb.Sp) in model group was significantly damaged (P<0.05), bone trabeculae were sparsely-arranged, discontinuous and discontinuous, fat droplets accumulated more, and osteogenic protein expression in bone tissue was significantly decreased (P<0.05). The mRNA and protein expression levels of EZH2 were significantly increased, while the mRNA and protein expression levels of Wnt3a and β-catenin were significantly decreased (P<0.05). Compared with the model group, the bone microstructure of rats in each administration group was significantly improved (P<0.05), the bone tissue pathological structure of rats in each administration group was improved to different degrees, the osteogenic protein expression in the bone tissue of rats in each administration group was increased to different degrees (P<0.05), and the EZH2 mRNA and protein expression levels in the bone tissue of rats in each administration group were significantly decreased. The mRNA and protein expression levels of Wnt3a and β-catenin were significantly increased (P<0.05). Conclusion Yu Rong Zhuanggu Gao can significantly improve bone loss in postmenopausal osteoporosis rats, and its mechanism is related to regulating EZH2/Wnt3a /β-catenin pathway to promote osteogenic differentiation.

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