摘要
目的观察生骨再造丸对激素性股骨头坏死(SONFH)大鼠H型血管生成及HIF-1α/VEGF信号通路的影响,探讨其骨修复作用的机制。方法采用甲泼尼龙琥珀酸钠联合脂多糖注射法进行SONFH造模,将50只大鼠分为空白组、模型组和生骨再造丸低、中、高剂量组,每组10只。生骨再造丸低、中、高剂量组分别予0.5、1.0、2.0 g/kg生骨再造丸药液灌胃4周,空白组和模型组予等量生理盐水灌胃。Micro-CT观察股骨头骨小梁结构及囊性变情况,免疫荧光染色检测股骨头内皮黏蛋白(Emcn)、CD31、Osterix表达,qRT-PCR和Western blot检测股骨头缺氧诱导因子(HIF)-1α、血管内皮生长因子(VEGF)及成骨相关转录因子Osterix、Runx2 mRNA和蛋白表达。结果与空白组比较,模型组大鼠股骨头骨小梁稀疏、断裂,软骨下囊性变形成,股骨头塌陷,股骨头H型血管标记物Emcn、CD31共染色面积和Osterix染色面积明显减少(P<0.01),HIF-1α、VEGF、Osterix、Runx2 mRNA和蛋白表达明显降低(P<0.01);与模型组比较,生骨再造丸各剂量组大鼠股骨头骨小梁稀疏、囊性变区域减少,生骨再造丸中、高剂量组大鼠股骨头Emcn、CD31共染色面积和Osterix染色面积明显增加(P<0.01),HIF-1α、VEGF、Osterix、Runx2 mRNA和蛋白表达明显升高(P<0.01)。结论生骨再造丸能通过上调HIF-1α/VEGF信号通路促进H型血管生成,进而促进骨修复,治疗SONFH。
Objective To explore the effects of Shenggu Zaizao Pills on promoting type-H vessels and on HIF-1α/VEGF signaling pathway of steroid-induced osteonecrosis of the femoral head(SONFH)rats;To explore the mechanism of its bone repair effect.Methods The SONFH rat model was made by injection of methylprednisolone sodium succinate combined with lipopolysaccharide.50 rats were divided into blank group,model group,Shenggu Zaizao Pills low-,medium-,and high-dosage groups,with 10 rats in each group.Shenggu Zaizao Pills low-,medium-,and high-dosage groups were given 0.5,1.0 and 2.0 g/kg Shenggu Zaizao Pills respectively for gavage for 4 weeks,and the other two groups were given the same amount of normal saline for gavage.Micro-CT was used to observe trabecular structure and cystic degeneration of femoral head,immunofluorescence staining was used to detect the expressions of femoral head endothelial mucin(Emcn),CD31 and Osterix,qRT-PCR and Western blot were used to detect femoral head hypoxia-inducible factor(HIF)-1α,vascular endothelial growth factor(VEGF),osteogenesisrelated transcription factors Osterix,Runx2 gene and protein expressions.Results Compared with the blank group,the femoral head bone trabeculae of the model group were sparse and broken,subchondral cysts were formed,the femoral head collapsed,the co-stained areas of femoral head type-H vascular markers Emcn and CD31,Osterix staining area were significantly reduced(P<0.01),the mRNA and protein expressions of HIF-1α,VEGF,Osterix,Runx2 significantly decreased(P<0.01).Compared with the model group,the sparse trabecular bone of femoral head of the rats in each dose group of Shenggu Zaizao pills and the cystic degeneration area was reduced,the co-staining area of Emcn and CD31,Osterix staining area of femoral head in Shenggu Zaizao Pills medium-and high-dosage groups increased significantly(P<0.01),the mRNA and protein expressions of HIF-1α,VEGF,Osterix,Runx2 significantly increased(P<0.01).Conclusion Shenggu Zaizao Pills can promote type-H vessles by up-regulating HIF-1α/VEGF signaling pathway,thereby promoting bone repair and treating SONFH.
作者
于海洋
卢增鹏
汪海燕
曹盼举
张虎林
郭成龙
梁恬
田杰祥
张晓刚
YU Haiyang;LU Zengpeng;WANG Haiyan;CAO Panju;ZHANG Hulin;GUO Chenglong;LIANG Tian;TIAN Jiexiang;ZHANG Xiaogang(Clinical College of Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China;Affiliated Hospital of Gansu University of Chinese Medicine,Lanzhou 730000,China;Chengdu University of Traditional Chinese Medicine,Chengdu 610000,China;Baoji Hospital of Chinese Medicine,Baoji 721000,China)
出处
《中国中医药信息杂志》
CAS
CSCD
2023年第5期91-96,共6页
Chinese Journal of Information on Traditional Chinese Medicine
基金
国家自然科学基金(81960832)
甘肃省自然科学基金(21JR11RA161)
甘肃省中医药管理局项目(GZKP-2021-17、GZKP-2020-31)
甘肃中医药大学院内课题(gzfy-2021-04)
宝鸡市卫生健康委员会科研项目(2020-026)
张晓刚全国名老中医药专家传承工作室建设项目(2022年)。
