斯赤列提取物抑制骨关节炎模型大鼠的异常血管新生  

Extracts of Sambucus adnata Wall.inhibit abnormal angiogenesis in a rat model of osteoarthritis

作　　者：江自鲜 陆玉春 李朝梦 郑梅梅 李秀芳 王文静 Jiang Zixian;Lu Yuchun;Li Chaomeng;Zheng Meimei;Li Xiufang;Wang Wenjing(Yunnan University of Chinese Medicine,Kunming 650500,Yunnan Province,China;NeuroDawn Pharmaceutical,Nanjing 210000,Jiangsu Province,China)

机构地区：[1]云南中医药大学,云南省昆明市650500 [2]南京宁丹新药技术有限公司,江苏省南京市210000

出　　处：《中国组织工程研究》2024年第34期5458-5466,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(82160744),项目负责人:王文静;云南省宋少江专家工作站(202305AF150030),项目负责人:王文静。

摘　　要：背景:课题组前期研究表明,斯赤列提取物具有促进成骨细胞增殖分化,促进骨折愈合、抗炎、抗氧化等作用,可有效缓解骨关节炎的发展。血管内皮生长因子则为骨关节炎严重程度评价的生物标记物。目的:探讨斯赤列两种提取物(甲醇提取物SAW-ME、二氯甲烷提取物SAW-DCE)对抗骨关节炎血管新生的作用及机制。方法:(1)采用前交叉韧带切断法复制大鼠骨关节炎模型(模型组),分别给予SAW-ME、SAW-DCE干预,并设假手术组为对照。运用免疫组化、免疫荧光等方法检测大鼠关节中血管内皮生长因子A、血清中血管内皮生长因子和软骨下骨H型血管的变化。(2)以血管内皮细胞EA.hy926为研究对象,给予SAW-ME、SAW-DCE干预,MTT法检测对细胞增殖的影响;采用血管内皮生长因子诱导EA.hy926细胞,复制血管新生模型,进行划痕实验、成管实验研究其作用和机制。(3)采用EA.hy926细胞进行转录组测序,分析给予SAW-DCE干预后细胞差异基因及相关功能的特征变化。结果与结论:(1)SAW-ME、SAW-DCE可下调骨关节炎大鼠膝关节软骨中血管内皮生长因子A的表达,减少关节中H型血管的生成;SAW-ME可显著降低骨关节炎大鼠血清中血管内皮生长因子水平(P <0.05);SAW-DCE可降低骨关节炎大鼠血清中血管内皮生长因子水平,但差异无显著性意义。(2)SAW-ME、SAW-DCE均可显著抑制血管内皮细胞迁移和成管,下调Ang1和Tie2蛋白的表达。(3)转录组测序分析发现骨关节炎血管异常新生与PI3K/AKT信号通路有关。(4)结果表明SAW-ME、SAW-DCE能够抑制骨关节炎模型大鼠异常血管新生,其作用机制可能与调控Ang1/Tie2、PI3K/AKT信号通路有关。BACKGROUND:Previous studies showed that extracts of Sambucus adnata Wall.have the ability to promote the proliferation and differentiation of osteoblasts,fracture healing,anti-inflammatory and antioxidant effects,which can effectively alleviate the development of osteoarthritis.Vascular endothelial growth factor,on the other hand,is a biomarker for the evaluation of osteoarthritis severity.OBJECTIVE:To investigate the effect and mechanism of two extracts of Sambucus adnata Wall.(methanol extract SAW-ME and dichloromethane extract SAW-DCE)on angiogenesis in osteoarthritis.METHODS:(1)Rat models of osteoarthritis were established using anterior cruciate ligament transection and given SAW-ME and SAW-DCE.A sham group was set as a control.Immunohistochemistry and immunofluorescence were used to detect the changes of articular vascular endothelial growth factor A in joint tissue and vascular endothelial growth factor and“H”type blood vessels in serum of osteoarthritis rats.(2)Vascular endothelial cells EA.hy926 were used as the research object and intervened with SAW-ME and SAW-DCE.Cell proliferation was then detected by MTT assay.Vascular endothelial growth factor was used to induce EA.hy926 cells,and the model of angiogenesis was replicated.Cell scratch assay and tube formation assay were performed to study the role and mechanism.(3)EA.hy926 cells were used for transcriptome sequencing to analyze the characteristic changes of cell differential genes and related functions after SAW-DCE intervention.RESULTS AND CONCLUSION:(1)SAW-ME and SAW-DCE downregulated the expression of vascular endothelial growth factor A in the rat knee cartilage and reduced the formation of“H”type vessels in osteoarthritis rats.SAW-ME could significantly decrease the level of vascular endothelial growth factor in serum of osteoarthritis rats(P<0.05).SAW-DCE could also decrease the level of vascular endothelial growth factor in serum of osteoarthritis rats,but there was no significant change.(2)Both SAW-ME and SAW-DCE significantly inhibi

关 键 词：斯赤列 甲醇提取物 二氯甲烷提取物 血管新生 骨关节炎 血管内皮生长因子 PI3K/AKT 转录组学 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R446