乌丹丸对寒凝血瘀型EMS模型大鼠内膜血管生成及细胞外调节蛋白激酶-血管内皮生长因子/基质金属蛋白酶9通路的影响  

Effects of Wudan Pills on Endometrial Angiogenesis and ERK-VEGF/MMP-9 Pathway in Rats with Endometriosis of Cold Congelation and Blood Stasis

作　　者：祝洁 薛晓鸥[1] 贺稚平[1] 严培嘉 张嘉蔚 孙海芸[1] ZHU Jie;XUE Xiao-ou;HE Zhi-ping;YAN Pei-jia;ZHANG Jia-wei;SUN Hai-yun(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijng 100700)

机构地区：[1]北京中医药大学东直门医院,北京100700

出　　处：《世界中西医结合杂志》2022年第4期645-651,共7页World Journal of Integrated Traditional and Western Medicine

基　　金：北京中医药大学青年教师项目(2019-jms-082);王子瑜教授全国名老中医传承工作室项目;北京中医药大学新奥基金“名老中医药专家经验挖掘学术研究重点项目”。

摘　　要：目的探索乌丹丸对寒凝血瘀型子宫内膜异位症(Endometriosis,EMS)大鼠内膜组织中微血管密度(Platelet endothelial cell adhesion molecule-1,CD31),细胞外调节蛋白激酶(Extracellular regulated protein kinases,ERK)-血管内皮生长因子(Vascular endothelial growth factor,VEGF)/基质金属蛋白酶9(Matrix metalloprotein-9,MMP-9)通路的影响。方法选取有规律动情周期的雌性SD大鼠,随机将49只大鼠分为假手术组、模型组、乌丹丸高、中、低剂量组(2.4、1.2、0.6 g/kg)、散结镇痛组(0.48 g/kg)和地诺孕素组0.2 mg/kg,每组各7只,冰水浴联合自体子宫内膜移植法建立寒凝血瘀型EMS,连续给药4周后处死取材。免疫组织化学染色检测大鼠异位内膜CD31阳性细胞表达,以免疫酶联吸附试验(Enzyme linked immunosorbent assay,ELISA)检测大鼠血清VEGF、MMP-9水平;以蛋白免疫印迹法(Western Blot,WB)检测大鼠异位子宫内膜组织ERK-VEGF/MMP-9通路相关蛋白表达。结果(1)与假手术组比较,模型组CD31阳性率,血清中VEGF、MMP-9水平,异位内膜组织ERK、VEGF、MMP-9蛋白表达均明显升高(P<0.05);(2)与模型组比较,乌丹丸各剂量组、散结镇痛组、地诺孕素组异位内膜CD31阳性率,血清中VEGF、MMP-9水平,异位内膜ERK1/2、VEGF、MMP-9蛋白表达均明显降低,且地诺孕素组<乌丹丸高剂量组<乌丹丸中剂量组<散结镇痛组<低剂量组(P<0.01,P<0.05)。结论乌丹丸可抑制寒凝血瘀型子宫内膜异位症模型大鼠内膜血管生成,可能是通过下调ERK-VEGF/MMP-9通路来实现。Objective To explore the effects of Wudan Pills on microvessel density(platelet endothelial cell adhesion molecule-1,CD31)andextracellular signal-regulated kinase(ERK)-vascular endothelial growth factor(VEGF)/matrix metalloproteinase-9(MMP-9)pathway in endometrium of rats with endometriosis(EMS)of cold congelation and blood stasis.Methods Female SD rats with regular estrous cycle were randomly divided into the sham operation group, model group, high-,medium-,and low-dose Wudan pill groups(2.4,1.2,0.6 g/kg),SanjieZhentong group(0.48 g/kg),and dienogest group(0.2 mg/kg),seven in each group.The EMS rats of cold congelation and blood stasis was modeled by autologous endometrium transplantationcombined with ice water bath.The rats were sacrificed after administration of four consecutive weeks, and samples were taken.The expression of CD31 positive cells in rat ectopic endometriumwas detected byimmunohistochemical staining, and the levels of serum VEGF and MMP-9 were detected by enzyme-linked immunosorbent assay(ELISA).The expression of ERK-VEGF/MMP-9 pathway related proteins in rat ectopic endometrium was determined by Western blot(WB).Results(1)Compared with those in the sham operation group, the expression of CD31 positive cells, the levels of serum VEGF and MMP-9 and the expression of ERK,VEGF and MMP-9 proteins in rat ectopic endometrium wereincreased in the model group(P<0.05).(2)Compared with the those in the model group, the expression of CD31 positive cells, the levels of serum VEGF and MMP-9,and theexpression of ERK1/2,VEGF and MMP-9 proteins in rat ectopic endometrium in the administration groups were lowered(P<0.05)in the order of dienogest group < high-dose Wudan pill group<medium-dose Wudan pill group<SanjieZhentong group<low-doseWudan pill group(P<0.01,P<0.05),and the decrease was dose-dependent among the Wudan pill groups.Conclusion Wudan pills can inhibit endometrial angiogenesis in EMS rats of cold congelation and blood stasis, which may be achieved by down-regulation of ERK-VEGF/MMP-9 pathway.

关 键 词：子宫内膜异位症 寒凝血瘀型 乌丹丸 CD31 ERK-VEGF/MMP-9 

分 类 号：R285.5[医药卫生—中药学]