基于H型血管生成益气化瘀方对去卵巢小鼠骨折愈合的机制研究  

Study on mechanism of Yiqi Huayu Formula promoting fracture healing in ovariectomized mice based on type H angiogenesis

作　　者：胡晓惠 孙康晖 过丽强 黄子裕 赵永见[1,2,3] 孙悦礼 赵东峰 王拥军[1,2] 施杞[1,2,3] 舒冰[2,3] HU Xiaohui;SUN Kanghui;GUO Liqiang;HUANG Ziy;ZHAO Yongjian;SUN Yue;ZHAO Dongfeng;WANG Yongjun;SHI Qi;SHU Bing(Longhua Hospital Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Institute of Spinal Diseases,Shanghai Institute of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;MOE Key Laboratory of Muscle and Bone Theory and Treatment,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院,上海200032 [2]上海中医药大学上海市中医药研究院脊柱病研究所,上海200032 [3]教育部筋骨理论与治法重点实验室,上海200032

出　　处：《上海中医药大学学报》2024年第6期75-81,共7页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82274555,81973876,81929004)。

摘　　要：目的:探讨益气化瘀方对去卵巢小鼠骨折愈合及H型血管生成的机制。方法:3月龄C57BL/6雌性小鼠96只,随机分为假手术组33只、模型组33只、益气化瘀方组30只。模型组与益气化瘀方组小鼠建立双侧去卵巢模型,假手术组小鼠仅切除卵巢周围部分脂肪组织。术后8周,所有小鼠接受左侧胫骨中段骨折术。骨折后给予小鼠益气化瘀方或0.9%NaCl溶液灌胃,每日1次,连续4周。取各组小鼠胫骨组织,Micro-CT扫描观察骨折愈合情况,HE染色观察骨痂组织形态学变化,三点弯曲实验检测小鼠胫骨最大荷载力,流式细胞术分析不同时间点各组小鼠骨痂组织中H型血管内皮细胞占总内皮细胞比例,免疫双荧光染色观察骨痂组织中H型血管分布。结果:骨折后第28天Micro-CT分析结果显示,假手术组与益气化瘀方组小鼠骨痂组织基本完成塑形,模型组骨痂仍处于塑形阶段。组织学染色结果显示,假手术组骨折局部骨皮质较为平整,模型组仍可见明显骨痂组织,益气化瘀方组仍有较小骨痂组织。益气化瘀方组小鼠愈合胫骨的最大荷载力较模型组显著提高。流式细胞术和免疫荧光染色结果显示,假手术组小鼠骨折局部组织中H型血管内皮细胞在总内皮细胞中的占比在骨折后第3天开始上升,直至第7天达到顶峰;模型组H型血管内皮细胞占比在骨折后第5天升高,且各时间点细胞占比均显著低于假手术组;益气化瘀方组小鼠骨折后第3天时细胞比例开始升高,早于模型组。与模型组相比,骨折后第3、5天时,益气化瘀方组小鼠骨折局部H型血管内皮细胞的占比和H型血管生成显著增加。结论:益气化瘀方能够提高去卵巢小鼠骨折愈合早期H型血管的生成,可能是其促进去卵巢小鼠骨折愈合的机制之一。Objective:To explore the mechanism of Yiqi Huayu Formula on fracture healing and type H angiogenesis in ovariectomized mice.Methods:963-month-old C57BL/6 female mice were randomly divided into sham operation group with 33 mice,model group with 33mice and Yiqi Huayu Formula group with 30 mice.Bilateral ovariectomized models were established in the model group and the Yiqi Huayu Formula group,while part of the adipose tissue around the ovary was removed in the mice of the sham operation group.Eight weeks after the operation,left mid-tibial fracture operation was performed in all mice.After fracture,mice in Yiqi Huayu Formula group were intragastrically administrated with Yiqi Huayu Formula once a day for 4 consecutive weeks,while mice in the sham operation group and model group were treated with equal volume of saline.The tibia tissue of mice in each group was collected,and the fracture healing was observed by Micro-CT scanning,and the histomorphological changes of callus tissue were observed by HE staining.The maximum loading force of tibia of mice was detected by three-point bending test,and the proportion of type H vascular endothelial cells in total endothelial cells of callus tissue of mice in each group at different time was analyzed by flow cytometry,and the distribution of type H vessels in callus tissue was observed by immunofluorescence double staining.Results:On the 28th day after fracture,Micro-CT analysis results showed that the callus tissue of mice in the sham operation group and Yiqi Huayu Formula group was basically completed shaping,while the callus in the model group was still in the shaping stage.The results of HE staining showed that the bone cortex of the fracture part in the sham operation group was relatively flat,while the callus tissue was still obvious in the model group,and small callus tissue was also observed in Yiqi Huayu Formula group.The maximum loading force of the healed tibia in Yiqi Huayu Formula group was significantly higher than that in the model group.The results of flow cyt

关 键 词：骨质疏松性骨折 去卵巢 益气化瘀方 H型血管 

分 类 号：R285.5[医药卫生—中药学]