隔药饼灸对子宫内膜异位症大鼠血管内皮生长因子及其受体的影响

Effect of Herbal-cake-separated moxibustion on macrophage phagocytosis and activation of VEGF/VEGFR pathway in endometriosis rats

目的:观察隔药饼灸对子宫内膜异位症(EMs)模型大鼠血管内皮细胞生长因子及其受体(VEGF/VEGFR)的影响,分析艾灸通过激活腹腔巨噬细胞吞噬功能治疗EMs的机制。方法:雌性SD大鼠随机分为假手术组、模型组、药物组和隔药饼灸组,每组30只。采用摘除子宫内膜法建立EMs模型。药物组用孕三烯酮(60 mg/kg)灌胃,每日1次,连续7 d;隔药饼灸组隔药饼灸"大椎"和"命门",每穴均灸6壮,每日1次,连灸7 d。治疗结束后第2天取各组大鼠异位子宫内膜标本,计算其异位内膜体积,并用HE染色观察病理形态。用酶联免疫吸附测定法检测各组大鼠腹腔液中血管内皮细胞生长因子A(VEGFA)、可溶性VEGFR(sVEGFR)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1、IL-2和IL-6的含量,并用显微镜观察腹腔液中巨噬细胞的形态,用无色孔雀绿比色法和MTT比色法检测腹腔液中巨噬细胞的吞噬能力及活力。结果:与假手术组比较,模型组大鼠出现异位灶,腹腔液中VEGFA、sVEGFR及TNF-α、IL-1、IL-2、IL-6的表达水平均显著升高(P<0.05),巨噬细胞吞噬能力及各时间点活力均降低(P<0.05)。与模型组比较,药物组和隔药饼灸组大鼠子宫内膜异位灶体积显著缩小(P<0.05),且隔药饼灸组更为明显(P<0.05),隔药饼灸组的血管生成与炎性细胞浸润显著改善;药物组和隔药饼灸组大鼠腹腔液中VEGFA、sVEGFR、TNF-α、IL-1、IL-2和IL-6表达水平降低(P<0.05),且隔药饼灸组降低更为明显(P<0.05);隔药饼灸组和药物组巨噬细胞吞噬能力及细胞活力均升高(P<0.05),且隔药饼灸组升高更显著(P<0.05)。与模型组及药物组比较,隔药饼灸组巨噬细胞数量增多,体积增大。结论:隔药饼灸能够抑制VEGF及VEGFR的活化,并降低EMs大鼠炎性反应,增强巨噬细胞的吞噬功能。

Objective To observe the effect of herbal-cake-separated moxibustion on changes of vascular endothelial growth factor(VEGF)/VEGF receptor(VEGFR)in endometriosis(EMs)model rats,so as to analyze the regulatory mechanism of herbal-cake-separated moxibustion on EMs.Methods SD rats were randomly divided into sham operation,model,herbal-cake-separated moxibustion,and medication groups,with 30 rats in each group.The EMs rats model was established by removing the endometrium.Herbal-cake-separated moxibustion was applied to“Dazhui”(GV14)and“Mingmen”(GV4)for 6 moxa-cones every time.Rats of the medication group was treated by gavage of gestrinone(60 mg/kg).All the treatments were conducted once daily for 7 days.The ectopic endometrial volume was calculated and the pathological condition was observed by HE staining.The levels of vascular endothelial growth factor A(VEGFA),soluble VEGFR(sVEGFR)and tumor necrosis factor-α(TNF-α),interleukin(IL)-1,IL-2 and IL-6 in peritoneal fluid were measured by ELISA.The morphology of macrophages in peritoneal fluid was observed under microscope.The phagocytic capacity and activity of macrophages were detected by colorless malachite green colo-rimetry and MTT colorimetry,respectively.Results Compared with the sham operation group,the volume of endometriosis increased significantly and showed obvious angiogenesis;the expression levels of VEGFA,sVEGFR,TNF-α,IL-1,IL-2 and IL-6 in rats peritoneal fluid were significantly increased(P<0.05).The number and volume of macrophages,and the phagocytic function and activity of macrophages(P<0.05)were all decreased.After the treatment,all the indexes mentioned above were all significantly reversed in the herbal-cake-separated moxibustion and medication groups(P<0.05),and the effects of herbal-cake-separated moxibustion were superior to the gestrinone(P<0.05).Conclusion Herbal-cake-separated moxibustion can inhibit the activation of VEGF/VEGFR,reduce the inflammatory response of EMs rats and enhance the phagocytosis of macrophages.