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[雷公藤多苷片对Ⅱ型胶原诱导性关节炎大鼠滑膜血管生成的影响]

[Effect of Tripterygium Glycosides Tablets on synovial angiogenesis in rats with type Ⅱ collagen induced arthritis].

作者信息

Wang Jing-Xia, Liu Chun-Fang, Li Yi-Qun, Su Xiao-Hui, Liu Li-Ling, Tian Ya-Ge, Wang Jin-Xia, Jia Ke-Xin, Lin Na

机构信息

Institute of Chinese Materia Medica,China Academy of Chinese Medicine Sciences Beijing 100700,China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2019 Aug;44(16):3441-3447. doi: 10.19540/j.cnki.cjcmm.20190625.401.

Abstract

To observe the effect of Tripterygium Glycosides Tablets on angiogenesis of rats with type Ⅱ collagen-induced arthritis( CIA) and on the tube formation of human umbilical vein endothelial cells( HUVEC) in vitro. The HUVEC were induced by 20 μg·L-1 vascular endothelial growth factor( VEGF) in vitro,and were treated with 0. 1,1,10 mg·L-1 Tripterygium Glycosides Tablets continuously for 7 hours. The numbers of branches of tube formation were measured. SD rats were immunized to establish CIA. CIA rats were treated with 9,18,36 mg·kg-1·d-1 Tripterygium Glycosides Tablets for 42 days. Histopathological examination( HE) was performed to observe the vascular morphology and vascular density in the synovial membrane of the inflamed joints. Immunohistochemistry and immunofluorescence were performed to observe the expression of platelets-endothelial cell adhesion molecule( CD31) and αsmooth muscle actin( αSMA) in synovial membrane. Immunohistochemistry and Western blot were performed to observe the expression of hypoxia-inducible factors 1α( HIF1α) and angiotensin 1( Ang1) in the synovial tissue. The results showed that the numbers of branches of tube formation of HUVEC induced by VEGF were improved,and declined significantly after treated by Tripterygium Glycosides Tablets. Compared with the normal group,the vascular density,CD31 positive expression,CD31 +/αSMA-immature and total vascular positive expression in the synovial membrane of the model group were significantly increased,and so as HIF1α and Ang1 in the synovium. Tripterygium Glycosides Tablets reduced the synovial vascular density and inhibited the positive expression of CD31,CD31+/αSMA-immature blood vessels and total vascular,but has no effect on CD31+/αSMA+mature blood vessels. Tripterygium Glycosides Tablets also inhibited the expression of HIF1α and Ang1 in synovial membrane of inflammatory joints. Our results demonstrated that Tripterygium Glycosides Tablets could inhibit the angiogenesis of synovial tissue in CIA rats and the tube formation of HUVEC,which is related to the down-regulation of HIF1α/Ang1 signal axis.

摘要

观察雷公藤多苷片对Ⅱ型胶原诱导性关节炎(CIA)大鼠血管生成及对体外培养的人脐静脉内皮细胞(HUVEC)管腔形成的影响。体外培养的HUVEC用20μg·L-1血管内皮生长因子(VEGF)诱导,并分别用0.1、1、10mg·L-1雷公藤多苷片连续处理7小时,测量管腔形成的分支数。将SD大鼠免疫建立CIA模型,CIA大鼠分别用9、18、36mg·kg-1·d-1雷公藤多苷片处理42天。进行组织病理学检查(HE)以观察炎症关节滑膜的血管形态和血管密度,采用免疫组织化学和免疫荧光法观察滑膜中血小板-内皮细胞黏附分子(CD31)和α平滑肌肌动蛋白(αSMA)的表达,采用免疫组织化学和蛋白质印迹法观察滑膜组织中缺氧诱导因子1α(HIF1α)和血管紧张素1(Ang1)的表达。结果显示,VEGF诱导的HUVEC管腔形成分支数增加,经雷公藤多苷片处理后显著减少。与正常组相比,模型组滑膜的血管密度、CD31阳性表达率、CD31+/αSMA-未成熟血管及总血管阳性表达率均显著升高,滑膜中HIF1α和Ang1也升高。雷公藤多苷片可降低滑膜血管密度,抑制CD31、CD31+/αSMA-未成熟血管及总血管的阳性表达,但对CD31+/αSMA+成熟血管无影响。雷公藤多苷片还可抑制炎症关节滑膜中HIF1α和Ang1的表达。结果表明,雷公藤多苷片可抑制CIA大鼠滑膜组织血管生成及HUVEC管腔形成,这可能与下调HIF1α/Ang1信号轴有关。

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