中国骨质疏松杂志

损伤胶囊通过Wnt3a/β-catenin通路调控BMSCs成骨分化改善绝经后骨质疏松研究

Sunshang Capsule ameliorates postmenopausal osteoporosis by modulation of BMSCs osteogenic differentiation through the Wnt3a/β-catenin pathway

DOI：10.3969/j.issn.1006-7108.2026.07.003

中文关键词: 损伤胶囊骨髓间充质干细胞绝经后骨质疏松

英文关键词:Sunshang Capsule bone marrow mesenchymal stem cells postmenopausal osteoporosis

基金项目:甘肃省自然科学基金项目（23JRRA1245）

作者	单位
陶瑜晶1 叶丙霖2* 谢芋涛1 王东霞1 王薛涛1 任毅1 杨建霞1 邵文鹤1 杨鹏程1	1.甘肃中医药大学,甘肃兰州 730030 2.甘肃省中医院,甘肃兰州 730050

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中文摘要:

目的探讨损伤胶囊通过调控骨髓间充质干细胞成骨分化及Wnt3a/β-catenin信号通路对绝经后骨质疏松症的改善作用。方法体外实验：将P2代大鼠BMSCs分为5组：空白一组（基础培养基+空白血清）、空白二组（基础培养基+空白血清+成骨诱导剂）、中药低/中/高剂量组（基础培养基+相应剂量含药血清+成骨诱导剂）。传代培养后，CCK-8法检测细胞增殖，茜素红染色观察矿化能力，RT-qPCR和Western blot检测Wnt3a、β-catenin及CyclinD1的表达水平。体内实验：将60只SPF级雌性SD大鼠随机分为假手术组和卵巢切除组。术后8周，将卵巢切除组分为对照组、雌二醇组及中药低、中、高剂量组。药物干预12周后取股骨进行分析：Micro-CT检测BMD、BV/TV、Tb.N、Tb.Th及Tb.Sp；HE染色观察骨小梁结构及脂肪空泡沉积；RT-qPCR和Western blot检测骨组织Wnt3a、β-catenin及CyclinD1的表达水平。结果体外实验结果：大鼠BMSCs冻存复苏后初期呈短梭形贴壁，培养第5天出现集落化生长，培养12 h出现贴壁，24 h转为均匀的尖梭形，4～5 d细胞融合度超80%，形成有序同向生长阵列。茜素红染色显示，中剂量组矿化结节数量显著高于其他组（P＜0.05）。中剂量组Wnt3a、β-catenin及CyclinD1的mRNA和蛋白表达较空白组上调（P＜0.05）。体内实验结果：对照组BMD、BV/TV、Tb.N较Sham组显著降低（P＜0.01），Tb.Sp显著升高（P＜0.01）；低、中、高剂量组及雌二醇组均能改善上述指标（P＜0.05），其中中剂量组与雌二醇组效果最显著。对照组骨小梁断裂伴脂肪空泡沉积，各干预组骨小梁连续性恢复，脂肪空泡减少。对照组Wnt3a、β-catenin及CyclinD1的mRNA和蛋白表达显著下调（P＜0.01），各治疗组表达均上调（P＜0.05），以中剂量组和雌二醇组最显著。结论损伤胶囊通过激活Wnt3a/β-catenin信号通路，促进BMSCs成骨分化增强骨形成能力，改善卵巢切除大鼠骨微结构退化。

英文摘要:

Objective To explore the ameliorative effects of Sunshang Capsule on postmenopausal osteoporosis via regulating the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and the Wnt3a/β-catenin signaling pathway. Methods In vitro, P2 rat BMSCs were divided into blank (basal medium + blank serum), induction (basal medium + blank serum + osteogenic inducer), and low/medium/high-dose drug groups (basal medium + drug-containing serum + osteogenic inducer). Cell proliferation (CCK-8), mineralization (Alizarin red staining), and Wnt3a/β-catenin/CyclinD1 expression (RT-qPCR/Western blotting) were assessed. In vivo, 60 SPF female SD rats were divided into sham and ovariectomized (OVX) groups. After 8 weeks, OVX rats were further divided into control, estradiol, and low/medium/high-dose drug groups. After 12 weeks of intervention, the femurs were analyzed with micro-CT (BMD, BV/TV, Tb.N, Tb.Th, Tb.Sp), HE staining (bone trabeculae, fat vacuoles), and Wnt3a/β-catenin/CyclinD1 expression (RT-qPCR/Western blotting). Results In vitro, BMSCs showed short shuttle-shaped adhesion initially. They formed colonies by day 5, uniformed needle-like morphology by 24h, and were confluent over 80% with ordered growth by 4-5 days. Cells in the medium-dose group exhibited the highest mineralized nodules (P<0.05) and up-regulated expression of Wnt3a/β-catenin/CyclinD1 mRNA/protein (P<0.05). In vivo, rats in the OVX control group had lower BMD/BV/TV/Tb.N (P<0.01) and higher Tb.Sp (P<0.01) vs. Those in the sham group. Rats in the treatment groups (low/medium/high-dose, estradiol) ameliorated these indices (P<0.05), with those in the medium-dose and estradiol groups showing optimal effects. HE staining revealed restored trabecular continuity and reduced fat vacuoles in treatment groups. Wnt3a/β-catenin/CyclinD1 expression was down-regulated in the OVX control (P<0.01) but was up-regulated in treatment groups (P<0.05), with the most significant effect in the medium-dose and estradiol groups. Conclusion Sunshang Capsule ameliorates postmenopausal osteoporosis by activating the Wnt3a/β-catenin pathway, promoting BMSC osteogenic differentiation and enhancing bone formation, thereby alleviating OVX-induced bone microstructural degradation.

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