活血通络胶囊对激素性股骨头坏死囊性变成血管修复的影响  

Effects of Huoxue Tongluo Jiaonang(活血通络胶囊)on angiogenesis repair of cystic lesions of steroid-induced osteonecrosis of the femoral head

作　　者：杨晓强 肖欢 田佳庆 方伟华 彭鹏 肖方骏 何伟 何敏聪 魏秋实 YANG Xiaoqiang;XIAO Huan;TIAN Jiaqing;FANG Weihua;PENG Peng;XIAO Fangjun;HE Wei;HE Mincong;WEI Qiushi(The Third Clinical Medical College of Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong,China;Bijie Traditional Chinese Medicine Hospital,Bijie 551700,Guizhou,China;The Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510378,Guangdong,China)

机构地区：[1]广州中医药大学第三临床医学院,广东广州510006 [2]毕节市中医医院,贵州毕节551700 [3]广州中医药大学第三附属医院,广东广州510378

出　　处：《中医正骨》2024年第9期40-48,共9页The Journal of Traditional Chinese Orthopedics and Traumatology

基　　金：国家自然科学基金项目(8227150353,82004392);广东省基础与应用基础研究基金项目(2023A1515010551);广东省中医骨伤研究院开放课题重点项目(GYH202101-01,GYH202101-04);毕节市科学技术局2022年度“揭榜挂帅”项目(毕科合重大专项[2022]1号);中国博士后科学基金第75批面上资助项目(2024M750819);中医证候全国重点实验室研究生项目(SKLKY2024A0003)。

摘　　要：目的:探讨活血通络胶囊对激素性股骨头坏死(steroid-induced osteonecrosis of the femoral head,SONFH)囊性变成血管修复的影响。方法:(1)将从全髋关节置换术中收集的一部分SONFH囊性变组织制备成单细胞悬液,采用单细胞转录组学分析囊性变组织中的细胞组分。将另一部分SONFH囊性变组织制成病理切片,采用苏木素-伊红染色法观察囊性变中血管形态,并采用免疫荧光法观察囊性变中血管内皮细胞特异性蛋白的表达情况。(2)分别按照每天20 mg·kg^(-1)、40 mg·kg^(-1)、80 mg·kg^(-1)体质量以浓度1 mg·mL^(-1)、2 mg·mL^(-1)、4 mg·mL^(-1)的活血通络胶囊溶液和20 mL·kg^(-1)生理盐水给大鼠灌胃,获取低、中、高剂量活血通络胶囊含药血清及空白血清。(3)取生长良好的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs),分为空白血清组和低、中、高剂量含药血清组,空白血清组细胞常规培养,低、中、高剂量含药血清组细胞分别采用低、中、高剂量活血通络胶囊含药血清干预。干预48 h后,通过细胞计数试剂盒-8法测定活血通络胶囊含药血清对HUVECs增殖能力的影响。(4)将HUVECs分为对照组、地塞米松组和活血通络组,对照组细胞常规培养,地塞米松组细胞采用地塞米松干预,活血通络组细胞采用地塞米松和中剂量活血通络胶囊含药血清干预。干预至细胞长满后进行细胞划痕实验,观察活血通络胶囊含药血清对HUVECs迁移能力的影响。(5)HUVECs分组与干预方法同(4),干预7 d后进行成血管实验,观察活血通络胶囊含药血清对HUVECs成血管能力的影响。结果:(1)SONFH囊性变组织中细胞组分分析结果。SONFH囊性变组织中存在内皮细胞及具有血管生成功能的内皮细胞亚群。(2)SONFH囊性变组织病理学观察结果。苏木素-伊红染色结果显示,SONFH囊性变组织中存在椭圆形的血管腔,血管内膜稍增厚、弹力纤维增生分�Objective:To explore the effects of Huoxue Tongluo Jiaonang(活血通络胶囊,HXTLJN)on angiogenesis repair of cystic lesions of steroid-induced osteonecrosis of femoral head(SONFH).Methods:①The tissues collected from the site of cystic lesions during the total hip arthroplasty in patients with SONFH were acquired,one part was made into single-cell suspension to analyze the cellular components in cystic lesions tissues based on the single cell transcriptomics,while,the other part was made into pathological sections to observe the vascular morphology and detect the expression of vascular endothelial cells(VECs)-specific proteins in cystic lesions tissues by using hematoxylin-eosin(HE)staining and immunofluorescence method,respectively.②Twelve rats were selected and ran domized into 4 groups.The rats in any three groups were intragastric administrated with HXTLJN solution at concentration of 1,2 and 4 mg/mL in daily dosage of 20,40 and 80 mg/kg,respectively,while,the ones in the last group with normal saline in daily dosage of 20 mL/kg,for making low-,medium-,and high-dose HXTLJN medicated serum and blank serum,respectively.③The well-growned human umbilical vein endothelial cells(HUVECs)were fetched and divided into blank serum group,and low-,medium-,and high-dose medicated serum groups.The HUVECs in blank serum group were cultured in the conventional Dulbecco’s Modified Eagle’s Medium(DMEM),while the ones in low-,medium-,and high-dose medicated serum groups were intervened with low-,medium-,and high-dose HXTLJN medicated serum,respectively.After 48-hour intervention,the effect of HXTLJN medicated serum on the proliferation ability of HUVECs was determined by using the cell counting kit-8(CCK8)assay.④The HUVECs were further divided into control group,dexamethasone(DEX)group and Huoxue Tongluo(活血通络,HXTL)group.The HUVECs in control group were cultured in the conventional DMEM,while the ones in DEX group were intervened with DEX,and the ones in HXTL group with DEX and medium-dose HXTLJN medicated

关 键 词：股骨头坏死 囊性变 成血管细胞 人脐静脉内皮细胞 活血通络 成血管修复 

分 类 号：R274.9[医药卫生—中西医结合]