健脾化湿通络方对佐剂关节炎大鼠滑膜血管新生的影响

目的观察健脾化湿通络方对佐剂关节炎大鼠滑膜血管PI3K/AKT/m TOR信号通路及缺氧诱导因子(HIF-1α)、血管内皮细胞生长因子A(VEGF-A)、滑膜微血管密度(MVD)、内皮抑素(ES)的影响。方法 50只大鼠随机均分成5组:正常对照组,模型对照组,甲氨蝶呤组,雷公藤多苷片组,健脾化湿通络方组。除正常对照组外,其余各组均采用弗氏完全佐剂建立佐剂关节炎大鼠模型。造模成功后,各治疗组给予相应药物灌胃,连续30 d。采用免疫组化法检测滑膜血管MVD表达,酶联免疫吸附法检测血清白细胞介素-6(IL-6)、IL-10、HIF-1α、VEGF-A表达,免疫印迹法检测滑膜血管PI3K、AKT、p-AKT、m TOR、ES蛋白表达。结果与正常对照组比较,模型对照组大鼠足跖肿胀度、关节炎指数升高,滑膜组织MVD计数升高,血清IL-6、VEGF、HIF-1α和滑膜血管PI3K、AKT、p-AKT、m TOR、ES表达显著升高,IL-10降低(P0.05或P0.01)。与模型对照组比较,健脾化湿通络方组MVD计数和血清VEGF-A、HIF-1α、IL-6表达降低,滑膜PI3K、p-AKT、m TOR、ES降低,血清IL-10升高。结论健脾化湿通络方通过调节PI3K/AKT/m TOR通路、HIF-1α及ES表达改善滑膜血管新生。

Effect of Jianpi Huashi Tongluo Formula on synovial angiogenesis in rats with adjuvant arthritis

Objective To observe the mechanism of Jianpi (invigorating the spleen)Huashi (resovling dampness)Tongluo (removing obstruction in collaterals)Formula (JHTF)on PI3K/AKT/mTOR signa-ling pathway,hypoxia inducible factor (HIF-1α),vascular endothelial growth factor (VEGF-A),mi-crovessel density (MVD)and endostatin (ES)of synovial tissue in rats with adjuvant arthritis (AA). Methods Fifty rats were randomly divided into five groups — normal control group,model group, methotrexate group,tripterygium glycosides tablet group and JHTF group.Rat AA model was induced by using Freund's complete adjuvant.After modeling,each treatment group were i.g.corresponding medica-tion for consecutive 30 days,and then the expression of microvascular density (MVD)was detected using immunohistochemical,interleukin (IL)-6,IL-10,HIF-1αand VEGF were measured by enzyme-linked immunosorbent assay,and PI3K,AKT1,p-AKT1,mTOR and ES proteins were detected by Western blotting.Results Compared with normal control group,footpad swelling,arthritic index increased,ser-um IL-6,VEGF and HIF-1αand MVD,PI3K,AKT1,p-AKT1,mTOR,ES in synovial vascular signifi-cantly increased in the model group(P <0.05 or P <0.01).Compared with the model group,the MVD count,serum VEGF-A,HIF-1αand IL-6 decreased in JHTF group.The expressions of PI3K,p-AKT1, mTOR,ES in synovial vascular in JHTF group were lower ,and serum IL-10 was higher than those of the model control group.Conclusion Jianpi Huashi Tongluo Formula can improve angiogenesis by regula-ting PI3K/AKT/mTOR pathway,expression of HIF-1αand ES in rat AA model.