目的观察补肾温阳化瘀方对子宫内膜异位症(endometriosis,EM)肾阳虚血瘀证模型大鼠缺氧诱导因子-1α(hypoxia inducible factor-1,HIF-1α)、脯氨酸羟化酶2(proline hydroxylase2,PHD2)、希佩尔-林道抑癌基因(von Hippel Lindau disease,VHL)表达的影响,探讨EM发病机制及补肾温阳化瘀方治疗EM的作用机制。方法将50只SD大鼠随机分为正常组、假手术组、模型组、中药组、西药组,每组10只。其中正常组、假手术组常规饲养,模型组、中药组、西药组采用“延长冰箱冷冻加冰水浸泡”30天,再结合“自体移植法”创建EM肾阳虚血瘀证模型。术后中药组给予补肾温阳化瘀方(3.33 g/mL)1mL灌胃,西药组予以孕三烯酮0.5 mg/(kg·d)灌胃,模型组、正常组、假手术组灌服等体积的生理盐水,共给药4周。观察大鼠异位灶大小及形态,放免法检测血清CA125、血浆环磷酸腺苷(cAMP)、血浆环磷酸鸟苷(cGMP)的表达,HE染色光镜观察子宫在位内膜及异位组织形态学的变化,免疫组化SABC法、Western blot法检测HIF-1α、PHD2、VHL蛋白表达及含量,RT-PCR方法检测HIF-1α、PHD2、VHL mRNA表达。结果模型组大鼠异位灶生长明显,中药组、西药组治疗后,异位灶体积明显缩小。与正常组及假手术组比较,模型组CA125及cGMP水平明显升高,cAMP水平明显降低(P<0.05);HIF-1αmRNA、蛋白表达及含量均升高(P<0.05),PHD2、VHL mRNA、蛋白表达及含量均降低(P<0.05)。与模型组比较,中药组及西药组CA125及cGMP水平明显降低,cAMP水平明显升高,HIF-1αmRNA、蛋白表达及含量均降低,PHD2、VHL mRNA、蛋白表达及含量均升高,差异均有统计学意义(P<0.05)。与中药组比较,西药组PHD2蛋白含量更高(P<0.05)。HIF-1α与PHD2呈负相关(r=-0.799,P=0.00),HIF-1α与VHL呈负相关(r=-0.625,P=0.003)。结论补肾温阳化瘀方可有效治疗EM,其作用机制可能与降低HIF-1α、CA125、cGMP含量,上调PHD2、VHL、cAMP表达有关。 Objective To observe the effect of Bushen Wenyang Huayu Recipe on the expression of hypoxia inducible factor-1 (Hypoxia inducible factor-1α) and proline in rats with kidney-yang deficiency and blood stasis syndrome model of endometriosis (EM) (PHD2) and von Hippel Lindau disease (VHL), and to explore the mechanism of EM and the mechanism of BushenYangyang Huayu Recipe in treating EM. Methods 50 SD rats were randomly divided into normal group, sham operation group, model group, TCM group and western medicine group, with 10 rats in each group. The normal group and the sham-operated group were fed routinely, the model group, the traditional Chinese medicine group and the western medicine group were treated with “Lengthening the refrigerator and ice-water immersion” for 30 days, then combined with “Autologous transplantation” model. The rats in the traditional Chinese medicine group were administrated 1ml of BushenYangyang Huayu Recipe (3.33 g / mL) orally, and the rats in the western medicine group were fed with 0.5 mg / (kg · d) of gestodene, and the model group, normal group and sham operation group Equal volume of saline for a total of 4 weeks. The size and morphology of heterotopic foci of rats were observed. The levels of serum CA125, cAMP and cGMP were detected by radioimmunoassay. The eutopic endometrium and ectopic tissues The changes of HIF-1α, PHD2 and VHL proteins were detected by immunohistochemical SABC and Western blot. The expressions of HIF-1α, PHD2 and VHL mRNA were detected by RT-PCR. Results In the model group, the heterotopic foci grew obviously. After the treatment, the volume of the heterotopic foci of the Chinese medicine group and the western medicine group decreased significantly. Compared with normal group and sham operation group, CA125 and cGMP levels in model group were significantly increased, cAMP level was significantly decreased (P <0.05), HIF-1α mRNA and protein expression and content were increased (P <0.05), PHD2, VHL mRNA , Protein expression and content were decreased (P <0.05). Compared with model group, the levels of CA125 and cGMP in Chinese medicine group and western medicine group were significantly decreased, the level of cAMP was significantly increased, the expression and content of HIF-1α mRNA and protein were decreased, and the mRNA and protein expressions and contents of PHD2 and VHL were increased Statistical significance (P <0.05). Compared with the traditional Chinese medicine group, the western medicine group PHD2 protein content was higher (P <0.05). HIF-1αwas negatively correlated with PHD2 (r = -0.799, P = 0.00), while HIF-1αwas negatively correlated with VHL (r = -0.625, P = 0.003). Conclusion BushenYangyang Huayu can effectively treat EM. The mechanism may be related to decreasing the content of HIF-1α, CA125 and cGMP and up-regulating the expression of PHD2, VHL and cAMP.