艾灸对佐剂性关节炎大鼠踝关节滑膜组织低氧诱导因子-1α和血管内皮生长因子表达的影响

Effect of moxibustion on the expressions of hypoxia inducible factor-1α and vascular endothelial growth factor in ankle synovial tissue of rats with adjuvant arthritis

目的:观察艾灸对佐剂性关节炎(AA)大鼠踝关节滑膜组织低氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)蛋白表达的影响,探讨艾灸抑制类风湿关节炎关节滑膜血管新生、改善关节症状的作用机制。方法:60只健康成年雄性SD大鼠随机分为正常组、模型组、艾灸组和甲氨蝶呤组,每组15只。采用右后足跖皮下注射弗氏完全佐剂法制备AA大鼠模型。艾灸组予双侧“足三里”“关元”和“阿是穴”艾灸治疗,每次20 min,每日1次,连续干预3周;甲氨蝶呤组予甲氨蝶呤(0.35 mg/kg)灌胃给药,每周2次,连续干预3周。采用足趾容积测量仪检测大鼠足跖容积;HE染色法观察大鼠踝关节滑膜组织病理情况;免疫组织化学法和Western blot法检测大鼠踝关节滑膜组织HIF-1α、VEGF蛋白表达情况。结果:与正常组比较,模型组大鼠的足趾容积和踝关节滑膜组织HIF-1α、VEGF蛋白表达均明显升高(P<0.01),滑膜组织出现明显增生且有大量新生血管形成;与模型组比较,艾灸组和甲氨蝶呤组大鼠的足趾容积和踝关节滑膜组织HIF-1α、VEGF蛋白表达均明显降低(P<0.05,P<0.01),滑膜组织未见明显增生,仅见少数新生血管形成;与甲氨蝶呤组比较,艾灸组大鼠足趾容积升高(P<0.05)。结论:艾灸能改善AA大鼠关节肿胀和抑制滑膜组织血管新生,其作用机制可能与调节踝关节滑膜组织HIF-1α、VEGF蛋白的表达有关。

Objective To observe the effect of moxibustion on the expressions of hypoxia inducible factor-1α(HIF-1α) and vascular endothelial growth factor(VEGF) in ankle synovial tissue of rats with adjuvant arthritis(AA),so as to explore the mechanism of moxibustion in inhibiting synovial angiogenesis and improving joint symptoms of rheumatoid arthritis.Methods Sixty healthy male SD rats were randomly divided into normal group,model group,moxibustion group and medication group,with 15 rats in each group.AA rat model was established by subcutaneous injection of Freund's complete adjuvant into the right hind paw.Rats in the moxibustion group were treated with “Zusanli”(ST36),“Guanyuan”(CV4) and “Ashi” point moxibustion,20 min each time,once a day,for consecutive 3 weeks.Rats in the medication group were given methotrexate(0.35 mg/kg) intragastric administration,twice a week,for consecutive 3 weeks.Foot plantar volume of rats was measured by toe volume mea-suring instrument.HE staining was used to observe the histopathology of ankle synovium.The protein expressions of HIF-1α and VEGF in ankle synovial tissue were detected by immunohistochemistry and Western blot.Results Compared with the normal group,the foot plantar volume and the protein expressions of HIF-1α and VEGF in synovial tissue of ankle joint were significantly increased(P<0.01) in the model group,the synovial tissue showed obvious hyperplasia and a large number of neovasculogenesis.Following the interventions,the foot plantar volume and the protein expressions of HIF-1α and VEGF in synovial tissue of ankle joint were significantly decreased(P<0.05,P<0.01) in both moxibustion and medication groups in contrast to the model group,and there was no obvious proliferation of synovial tissue,and only a few neovascularization was observed.Compared with the medication group,the foot plantar volume was decreased(P<0.05) in the moxibustion group.Conclusion Moxibustion can improve joint swelling and inhibit synovial angiogenesis in AA rats,and its mechanism may be related to down-regulating of HIF-1α and VEGF protein expressions.