体外冲击波治疗通过激活Wnt5a/Ca2+信号通路改善大鼠股骨牵张成骨的分子机制

doi:10.3969/j.issn.1007-3205.2024.01.009

摘要/Abstract

摘要： 目的探究体外冲击波治疗通过激活Wnt5a/Ca2+信号改善大鼠股骨牵张成骨的机制。
方法 60只8周龄Sprague Dawley雄性大鼠分为对照组、股骨牵张模型组和体外冲击波疗法（extracorporeal shock wave therapy，SWT）组（n=20）。使用Cat Walk XT系统测量大鼠步态参数。通过显微CT（Micro-CT）扫描分析大鼠股骨形态。组织病理学分级采用Mankin评分标准，并分析软骨下骨各区域Wnt5a阳性细胞百分比。实时聚合酶链反应分析大鼠股骨Ⅰ型胶原和骨桥蛋白mRNA表达。免疫组织化学染色分析骨形态发生蛋白2（bone morphogenetic protein 2，BMP2）、血管内皮生长因子（vascular endothelial growth factor，VEGF）和增殖细胞核抗原（proliferating cell nuclear antigen，PCNA)的表达。蛋白印迹分析Wnt5a/Ca2+信号通路蛋白表达。
结果模型组摆动速度、最大接触面积、单站姿、占空比和支架摆动时间低于对照组，ESWT组摆动速度、最大接触面积、单站姿、占空比和支架摆动时间高于模型组（P＜0.05）。模型组骨小梁体积分数、小梁厚度和骨密度小于对照组，ESWT组骨小梁体积分数、小梁厚度和骨密度大于模型组（P＜0.05）。模型组股关节病理组织学（Mankin）评分高于对照组，Wnt5a阳性细胞数量少于对照组，ESWT组股关节Mankin评分低于模型组，Wnt5a阳性细胞数量多于模型组（P＜0.05）。与对照组相比，模型组大鼠软骨下骨板浅表区可见骨细胞陷窝，ESWT组股关节面损伤程度明显减轻（P＜0.05）。ESWT诱导模型大鼠Wnt5a标记明显增加（P＜0.05）。模型组Ⅰ型胶原和骨桥蛋白mRNA表达低于对照组，ESWT组Ⅰ型胶原和骨桥蛋白mRNA表达高于模型组（P＜0.05）。模型组BMP2、VEGF和PCNA染色阳性细胞少于对照组，ESWT组BMP2、VEGF和PCNA染色阳性细胞多于模型组（P＜0.05）。模型组CaMKⅡ、PLC、Wnt5a蛋白表达高于对照组，PKC蛋白表达高于对照组，ESWT组CaMKⅡ、PLC、Wnt5a蛋白表达低于模型组，PKC蛋白表达低于模型组（P＜0.05）。
结论体外冲击波治疗通过激活Wnt5a/Ca2+信号通路促进大鼠股骨牵张成骨新生血管形成、细胞增殖和成骨生长因子的表达，改善骨力学性能和骨矿化加速。

关键词: 股骨, 骨生成, 牵张, 体外冲击波疗法

Abstract: Objective To explore the mechanism of extracorporeal shock wave therapy (ESWT) to improve the distraction osteogenesis of rat femur by activating Wnt5a/Ca2+ signal.
Methods Sixty 8-week-old Sprague Dawley male rats were divided into a control group, a femoral traction model group, and an ESWT group (n=20). Rat gait parameters were measured using the CatWalk XT system, and the morphology of rat femur was analyzed through Micro-CT scanning. The histopathological grading was performed using the Mankin scoring standard, and the percentage of Wnt5a positive cells in each region of the subchondral bone was analyzed. Real-time polymerase chain reaction (RT-PCR) was used to analyze type Ⅰ collagen and osteopontin mRNA expression in rat femur. Immunohistochemical staining was used to analyze the expression of bone morphogenetic protein 2 (BMP2), vascular endothelial growth factor (VEGF), and proliferating cell nuclear antigen (PCNA). Western blot was used to analyze Wnt5a/Ca2+ signaling pathway protein expression.
Results The swing speed, maximum contact area, single standing posture, duty cycle and stent swing time of rats in the model group were lower than those in the control group, while the swing speed, maximum contact area, single standing posture, duty cycle and stent swing time of rats in the ESWT group were higher than those in the model group (P＜0.05). The trabecular volume fraction, trabecular thickness and bone mineral density of rats were lower in the model group than in the control group, but higher in the ESWT group than in the model group (P＜0.05). The Mankin score of the femoral joint in the model group was higher than that in the control group, while the number of Wnt5a positive cells in the model group was lower than that in the control group; The Mankin score of the femoral joint in the ESWT group was lower than that in the model group, while the number of Wnt5a positive cells in the ESWT group was higher than that in the model group (P＜0.05). Compared with the control group, osteocyte lacunae could be seen in the superficial area of subchondral bone plate in the model group, and the degree of injury on the femoral joint surface in the ESWT group was significantly reduced (P＜0.05). The Wnt5a marker of ESWT-induced model rats increased significantly (P＜0.05). The expression of type Ⅰ collagen and osteopontin mRNA was lower in the model group than in the control group, but higher in the ESWT group than in the model group (P＜0.05). Compared with the control group, the positive cells of BMP2, VEGF and PCNA staining in the model group decreased, while the positive cells of BMP2, VEGF and PCNA staining in the ESWT group increased, as compared with the model group (P＜0.05). The expression of CaMKⅡ, PLC and Wnt5a protein and the expression of PKC protein in the model group were higher than those in the control group, while the expression of CaMKⅡ, PLC and Wnt5a protein and the expression of PKC protein in the ESWT group were lower than those in the model group (P＜0.05).
Conclusion ESWT can promote the formation of angiogenesis, cell proliferation and the expression of osteoblast growth factors by activating the Wnt5a/Ca2+ signaling pathway, improve the bone mechanical performance and accelerate bone mineralization.

Key words: Femur, Osteogenesis, Distraction, Extracorporeal shock wave therapy

黄康斌1, 陈曾凤2, 陈从山1, 张远洋1, 邱志伟1. 体外冲击波治疗通过激活Wnt5a/Ca2+信号通路改善大鼠股骨牵张成骨的分子机制 [J]. 河北医科大学学报, 2024, 45(1): 40-46.

HUANG Kang-bin1, CHEN Zeng-feng2, CHEN Cong-shan1, ZHANG Yuan-yang1, QIU Zhi-wei1. Molecular mechanism of extracorporeal shock wave therapy improving the distraction osteogenesis of rat femur by activating Wnt5a/Ca2+ signal pathway [J]. Journal of Hebei Medical University, 2024, 45(1): 40-46.

[1]	郑占乐, 刘家伦, 常恒瑞, 陈伟, 连晓东, 张英泽. 胫骨平台前1/5双反牵引在Hoffa骨折治疗中的初步临床应用 [J]. 河北医科大学学报, 2024, 45(1): 6-8.
[2]	丁辉海1, 崔建国2, 韩成相1. 血清TM、P-selectin及血液流变学相关指标预测老年股骨转子间骨折术后深静脉血栓发生风险的价值 [J]. 河北医科大学学报, 2024, 45(1): 29-34.
[3]	马晨, 寇文冠, 柯磊, 刘东帅, 张月震. 关节镜下应用氨甲环酸联合富血小板血浆注射治疗髋关节撞击综合征的效果 [J]. 河北医科大学学报, 2023, 44(9): 1032-1036.
[4]	龙玉斌, 李一然, 陈伟, 贾红伟, 韩宝君, 安明. 天玑机器人辅助股骨颈动力交叉钉系统固定治疗中老年股骨颈骨折的近期效果分析 [J]. 河北医科大学学报, 2023, 44(5): 541-546.
[5]	胡欣怡（综述）, 荣晓凤（审校）. 中轴型脊柱关节炎的病理变化与生物标志物 [J]. 河北医科大学学报, 2023, 44(11): 1360-1364.
[6]	郑占乐, 陈伟, 张弢, 李会杰, 王娟, 张英泽 . 取梁换柱部分置换术在股骨头坏死中的应用 [J]. 河北医科大学学报, 2023, 44(1): 112-115.
[7]	雷凯, 任荣, 李得春, 白景棠, 张学斌. 高海拔地区股骨颈骨折空心螺钉理想构型的三维测量研究 [J]. 河北医科大学学报, 2022, 43(9): 1088-1090.
[8]	吴少坪, 黄小玉, 吴丽苹, 张永发. 不同浓度罗哌卡因PENG阻滞用于老年股骨头置换术后多模式镇痛的效果 [J]. 河北医科大学学报, 2022, 43(12): 1407-1411,1422.
[9]	陈宇峰（综述）, 任栋, 王鹏程（审校）. Pauwels Ⅲ型股骨颈骨折内固定选择策略的研究进展[J]. 河北医科大学学报, 2022, 43(1): 108-113.
[10]	胡一平, 刘晓晖, 蔡福金, 唐可, 蒋家耀. 不同手术方式对移位性股骨颈骨折患者髋关节活动度、免疫炎性反应及凝血-纤溶系统的影响[J]. 河北医科大学学报, 2021, 42(9): 1046-1051.
[11]	叶丹丹, 朱燕宾, 陈伟, 张英泽. 股骨头、颈及转子间骨小梁三角的研究[J]. 河北医科大学学报, 2021, 42(9): 1093-1096.
[12]	王建华, 林艳, 吴永东, 王有宝. 经皮撬拨技术治疗粗隆间难复性骨折的效果及安全性研究[J]. 河北医科大学学报, 2021, 42(2): 145-149,153.
[13]	高尚聚, 李文毅, 张隆, 李旭明, 魏景超, 曹参. FHR、PFNA治疗伤前日常生活能力较好的高龄IFF患者效果对比研究[J]. 河北医科大学学报, 2021, 42(1): 34-37,59.
[14]	张合1，曹成明2，兰志恒1，赵志光1，刘巍1，韩守江1*. 改良单图像法手工测量髌骨不稳患者胫骨结节-股骨滑车沟间距的可行性分析[J]. 河北医科大学学报, 2020, 41(6): 660-664.
[15]	姚柯炜，张流畅. 股骨外侧壁厚度对于股骨近端骨折术后外侧壁骨折的预测价值[J]. 河北医科大学学报, 2020, 41(6): 738-741.

体外冲击波治疗通过激活Wnt5a/Ca2+信号通路改善大鼠股骨牵张成骨的分子机制

Molecular mechanism of extracorporeal shock wave therapy improving the distraction osteogenesis of rat femur by activating Wnt5a/Ca2+ signal pathway

HTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价