补肾活血汤对子宫内膜异位症大鼠Nrf2/Keap1通路的调节作用研究

Regulatory effect of Bushen Huoxue Decoction on Nrf2/Keap1 pathway in rats with endometriosis

关键词:  补肾活血汤  子宫内膜异位症  Nrf2/Keap1通路

Key words:  Bushen Huoxue Decoction  endometriosis  Nrf2/Keap1 pathway

摘要:

[目的] 研究补肾活血汤对子宫内膜异位症大鼠Nrf2/Keap1通路的调节作用。[方法] 90只大鼠随机分为假手术组、模型组、补肾活血汤低、中、高剂量组及达那唑组，15只/组。采用自体子宫内膜移植法建立子宫内膜异位症模型大鼠，补肾活血汤低、中、高剂量组按照2.5、5、10 g/kg灌胃给予补肾活血汤，达那唑组给予达那唑(0.02 g/kg)，每日1次，连续干预4周，采用酶联免疫吸附法(ELISA)检测大鼠血清雌二醇(E2)、癌抗原(CA)125水平及腹腔灌洗液白介素-6(IL-6)、白介素-8(IL-8)、前列腺素F-2α(PGF-2α)水平，苏木精-伊红(HE)染色法检测大鼠子宫组织病理学变化，实时聚合酶链式反应(PCR)检测子宫内膜异位症大鼠异位子宫组织Nrf2、Keap1 mRNA水平，蛋白印迹法(Westrn blot)测定大鼠异位子宫组织Nrf2、Keap1蛋白水平。[结果] 与假手术组大鼠相比，模型组大鼠血清E2、CA125水平及腹腔灌洗液IL-6、IL-8、PGF-2α水平升高(P<0.05)，异位子宫组织Nrf2 mRNA及蛋白水平升高(P<0.05)，Keap1 mRNA及蛋白水平降低(P<0.05)；与模型组大鼠相比，补肾活血汤各剂量组大鼠血清E2、CA125水平及腹腔灌洗液IL-6、IL-8、PGF-2α水平降低(P<0.05)，异位子宫组织Nrf2 mRNA及蛋白水平降低(P<0.05)，Keap1 mRNA及蛋白水平升高(P<0.05)，异位子宫内膜增厚等病理症状减轻。[结论] 补肾活血汤能够降低大鼠血清雌二醇及CA125水平，缓解子宫内膜异位症大鼠腹腔炎症水平，其机制可能与调节Nrf2/Keap1通路有关。

Abstract:

[Objective] To study the regulatory effect of Bushen Huoxue Decoction on Nrf2/Keap1 pathway in rats with endometriosis.[Methods] The 90 rats were randomly divided into sham operation group,model group,low,middle and high dose Bushen Huoxue Decoction groups and danazol group. Endometriosis model rats were established by autologous endometrial transplantation. Bushen Huoxue Decoction low,middle and high dose groups were given Bushen Huoxue Decoction according to 2.5,5 and 10 g/kg,and danazol group was given danazol (0.02 g/kg) once a day. After continuous intervention for 4 weeks,the levels of estradiol (E2) and cancer antigen (CA) 125 in serum and the levels of interleukin-6 (IL-6),interleukin-8 (IL-8) and prostaglandin F2α (PGF-2α) in peritoneal lavage fluid were detected by enzyme-linked immunosorbent assay (ELISA). The histopathological changes of uterus were detected by hematoxylin-eosin (HE) staining. The levels of Nrf2 and Keap1 mRNA in ectopic uterine tissue of rats with endometriosis were detected by real-time polymerase chain reaction (PCR). The levels of Nrf2 and Keap1 in ectopic uterine tissue of rats were measured by Westrn blot.[Results] Compared with the sham operation group,in model group,the levels of serum E2 and CA125 and the levels of IL-6,IL-8 and PGF-2 α in peritoneal lavage fluid were increased (P<0.05),Nrf2 mRNA and protein in ectopic uterus were increased (P<0.05),Keap1 mRNA and protein were decreased(P<0.05). Compared with the model group,the levels of E2 and CA125 in serum and the levels of IL-6,IL-8 and PGF-2 α in peritoneal lavage fluid,the levels of Nrf2 mRNA and protein in ectopic uterus were decreased(P<0.05),the levels of Keap1 mRNA and protein were increased(P<0.05),and the pathological symptoms such as thickening of ectopic endometrium alleviated in Bushen Huoxue Decoction groups.[Conclusion] Bushen Huoxue Decoction can reduce the levels of serum estradiol and CA125 and relieve abdominal inflammation in rats with endometriosis. The mechanism may be related to the regulation of Nrf2/Keap1 pathway.