甘草附子汤治疗佐剂性关节炎模型小鼠的抗滑膜血管生成机制研究-中国药房

甘草附子汤治疗佐剂性关节炎模型小鼠的抗滑膜血管生成机制研究

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篇名：	甘草附子汤治疗佐剂性关节炎模型小鼠的抗滑膜血管生成机制研究
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摘要：	目的：考察甘草附子汤通过抗滑膜血管生成途径对佐剂性关节炎（AA）模型小鼠的治疗作用机制。方法：取雄性Balb/c小鼠48只随机分为正常组、模型组、甘草附子汤组和雷公藤多苷组（阳性药物组），每组12只。除正常组外，其余小鼠左后足趾皮内一次性注射弗氏完全佐剂以建立AA模型。造模12 d后，正常组、模型组小鼠灌胃相应体积的水，甘草附子汤组（7.8 g/kg，以总生药量计算）和雷公藤多苷组（0.01 g/kg）小鼠灌胃相应药液20 mL/kg，每天1次，连续18 d。观察并记录小鼠关节病变情况，并采用水容积法测量小鼠足肿胀度；采用苏木精-伊红染色法观察小鼠踝关节组织病理学变化；采用免疫荧光法检测小鼠踝关节滑膜组织中血小板-内皮细胞黏附分子（CD31）、血管内皮生长因子（VEGF）的蛋白表达水平；采用Western blotting法检测小鼠踝关节滑膜组织中核转录因子κB（NF-κB）、锌指转录因子GATA4（GATA4）的蛋白表达水平。结果：与正常组比较，模型组小鼠足跖明显红肿，足肿胀度显著升高（P＜0.05）；踝关节滑膜组织增生，血管翳明显增多，可见大量炎症细胞及关节侵蚀等；滑膜组织中CD31、VEGF、NF-κB、GATA4的蛋白表达水平均显著升高（P＜0.05）。与模型组比较，甘草附子汤组小鼠足跖红肿现象减轻，足肿胀度显著降低（P＜0.05）；踝关节滑膜组织中血管翳有所减少，其他病理症状有所改善；滑膜组织中CD31、VEGF、NF-κB、GATA4的蛋白表达水平均显著降低（P＜0.05）。结论：甘草附子汤可通过下调关节滑膜组织中VEGF、NF-κB、GATA4蛋白的表达，减少滑膜组织中血管翳的形成并有效抑制血管新生，从而对AA模型小鼠发挥防治骨质破坏、保护关节等作用。
ABSTRACT：	OBJECTIVE： To investigate the therapeutic effect of Licorice and aconite decoction in the treatment of adjuvant arthritis （AA） model mice through anti-synovial angiogenesis pathway. METHODS： Totally 48 male Balb/c mice were randomly divided into normal group， model group， Licorice and aconite decoction group and tripterygium glycosides group （positive drug group）， with 12 mice in each group. Except for normal group， AA model was established by intradermal injection of Freund’s complete adjuvant into the left hind toe of mice. 12 d after modeling， normal group and model group were given same volume of water intragastrically； Licorice and aconite decoction group （7.8 g/kg，by total amount of crude drug） and tripterygium glycosides group （0.01 g/kg） were given relevant medicine 20 mL/kg intragastrically， once a day， for consecutive 18 d. The joint lesions of mice were observed and recorded， and the foot swelling degree of mice was measured by water volume method. HE staining was used to observe the pathological change of ankle joint in mice. The protein levels of platelet endothelial cell adhesion molecule-1 （CD31） and vascular endothelial growth factor （VEGF） were determined by immunofluorescence assay. The protein expressions of nuclear factor κB（NF-κB） and zinc finger transcription factor GATA4 （GATA4） were detected by Western blotting assay. RESULTS： Compared with normal group， ankle joint of model group mice was markedly reddened and swollen， and foot swelling degree increased significantly （P＜0.05）. Synovial tissue of ankle joint proliferated， pannus increased significantly， and a large number of inflammatory cells and joint erosion were observed. The protein expression of CD31， VEGF， NF-κB and GATA4 in synovial tissue of mice were increased significantly （P＜0.05）. Compared with model group， the redness and swelling of ankle joint in mice were alleviated， and the foot swelling degree was significantly reduced in Licorice and aconite decoction group （P＜0.05）. Pannus in synovial tissue decreased and other pathological symptoms were improved. The protein expression of CD31， VEGF， NF-κB and GATA4 were decreased significantly in synovial tissue （P＜0.05）. CONCLUSIONS： Licorice and aconite decoction can decrease the protein expression of VEGF，NF-κB and GATA4 in synovial tissue， reduce the generation of pannus in synovial tissue and effectively inhibit the angiogenesis in synovial tissue so as to prevent bone destruction and protect joint of AA model mice.
期刊：	2019年第30卷第12期
作者：	蔡悦，刘晓龙，郭静，张囡
AUTHORS：	CAI Yue，LIU Xiaolong，GUO Jing，ZHANG Nan
关键字：	甘草附子汤；类风湿关节炎；佐剂性关节炎；血管新生；血小板-内皮细胞黏附分子；血管内皮生长因子；锌指转录因子GATA4；核因子κB；小鼠
KEYWORDS：	Licorice and aconite decoction； Rheumatoid arthritis； Adjuvant arthritis； Angiogenesis； CD31； VEGF； GATA4； NF-κB； Mice
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