萝卜硫素对大鼠子宫内膜异位组织Nrf2/Keap1/HO-1信号通路影响

The study on the effect of sulforaphane on nuclear factor E-related factor 2/Kelch liked ECH-associated protein 1/heme oxygenase-1 signaling pathway in ectopic endometrial tissues of rats with endometriosis

目的:探讨萝卜硫素(SFN)对子宫内膜异位症(EMs)大鼠异位组织核因子E2相关因子2(Nrf2)/Kelch样ECH相关蛋白(Keap1)/血红素氧合酶-1(HO-1)信号通路的影响。方法:将造模成功的雌性SD大鼠随机分为模型组、阳性对照组(0.5 mg/kg,孕三烯酮胶囊)及低(5 mg/kg)、中(10 mg/kg)、高剂量(15 mg/kg)SFN组,每组8只;此外,设立空白对照(假手术)组。各组均给药8周后处死大鼠,计算子宫内膜异位病灶体积和粘连评分。实时荧光定量PCR技术检测Nrf2和HO-1的mRNA表达,免疫印迹法检测Nrf2、Keap1和HO-1蛋白表达。结果:与模型组相比,3种剂量SFN均能有效抑制EMs大鼠病灶体积和粘连评分,并呈剂量依赖趋势(P<0.05)。此外,与空白对照组相比,模型组大鼠内膜组织中Nrf2、HO-1 mRNA表达水平以及Nrf2、Keap1、HO-1蛋白表达减少(P<0.05)。给予SFN后,Nrf2、HO-1 mRNA表达水平以及Nrf2、Keap1、HO-1蛋白表达均升高,并呈浓度依赖性(P<0.05)。结论:Nrf2/Keap1/HO-1信号通路参与EMs发生;SFN可能通过激活Nrf2/Keap1/HO-1信号通路,上调HO-1等抗氧化蛋白的表达而起到治疗EMs的效果。

Objective:To investigate the effect of sulforaphane(SFN)on nuclear factor E2-related factor 2(Nrf2)/Kelch liked ECH-associated protein 1(Keap1)/heme oxygenase-1(HO-1)signaling pathway in ectopic endometrial tissues of rats with endometriosis(EMs).Methods:The successfully modeled female SD rats with Ems were randomly divided into 5 groups(8 rats in each group),which included model group,positive control group(0.5 mg/kg gestrinone capsule),low-dose SFN group(5 mg/kg SFN),medium-dose SFN group(10 mg/kg SFN),and high-dose SFN group(15 mg/kg SFN).In addition,the blank control group(rats with sham operation)was also established.The rats in the model group and the blank control group were given the same volume of 0.9%NaCl solution.The rats in the 5 groups were sacrificed after the 8 weeks administration.The endometriotic lesion volume and adhesion score of the rats in the 5 groups were calculated.The mRNA expressions of Nrf2 and HO-1 of these rats were detected by Real-time PCR,and the expressions of Nrf2,Keap1 and HO-1 protein of these rats were detected by Western Blot.Results:Compared with those of the rats in the model group,the lesion volume and adhesion score of rats with Ems in the three doses SFN groups had been inhibited effectively,and which had showed a dose-dependent trend(P<0.05).In addition,compared with those of the rats in the blank control group,the expressions levels of Nrf2,HO-1 mRNA,and Nrf2,Keap1,and HO-1 protein in the endometrial tissue of the rats in the model groups had decreased significantly(P<0.05).After administration of SFN,the expressions of Nrf2,HO-1 mRNA,Nrf2,Keap1,and HO-1 protein of the rats with EMs had increased significantly,and which had showed a concentration-dependent trend(P<0.05).Conclusion:The Nrf2/Keap1/HO-1 signaling pathway is involved in the occurrence of endometriosis of rats.SFN may play a role in the treatment of endometriosis by activating Nrf2/Keap1/HO-1 signaling pathway and up-regulating the expression of antioxidant proteins,such as HO-1.