汉黄芩素调节SIRT1/Nrf2信号通路对子宫内膜异位症大鼠铁死亡的影响

Impact of wogonin on ferroptosis in endometriosis rats by regulating SIRT1/Nrf2 signaling pathway

目的探讨汉黄芩素(Wog)对子宫内膜异位症(EM)大鼠铁死亡的影响及其作用机制。方法构建EM大鼠模型,将SD大鼠分为空白组(CT组)、EM模型组(EM组)、Wog低剂量组(Wog L组)、Wog高剂量组(Wog H组)、Wog+沉默调节蛋白1(SIRT1)抑制剂(EX527)组。给药4周后,检测异位病灶体积,ELISA测定血清雌二醇(E2)、孕激素(P)、白介素-1β(IL-1β)、IL-6的含量,HE染色观察异位内膜组织病理损伤,普鲁士蓝染色检测铁离子聚集情况,铁离子比色法检测Fe^(2+)浓度,Western blot检测SIRT1、核因子E2相关因子2(Nrf2)、谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLC7A11)和铁蛋白轻链(FTL)蛋白表达水平。结果与CT组相比,EM组大鼠异位内膜组织腺体较少,大量炎性细胞浸润和铁离子沉积,异位病灶体积、E2、P、IL-1β、IL-6、Fe^(2+)水平升高,SIRT1、Nrf2、GPX4、FTL和SLC7A11蛋白表达水平下降(P<0.05);与EM组相比,Wog L、Wog H组异位内膜组织腺体增加,离子沉积少见,异位病灶体积、E2、P、IL-1β、IL-6、Fe^(2+)水平降低,SIRT1、Nrf2、GPX4、FTL和SLC7A11蛋白表达水平升高(P<0.05);与Wog H组相比,Wog+EX527组大鼠异位内膜组织炎性细胞浸润和铁离子沉积增加,异位病灶体积、E2、P、IL-1β、IL-6、Fe^(2+)水平升高,SIRT1、Nrf2、GPX4、FTL和SLC7A11蛋白表达水平下降(P<0.05)。结论Wog可能通过激活SIRT1/Nrf2信号通路抑制EM大鼠铁死亡。

Objective To investigate the impact of wogonin(Wog)on ferroptosis in endometriosis(EM)rats and its mechanism.Methods An EMrat model was constructed;SD rats were grouped into blank group(CT group),EMmodel group(EM group),Wog low-dose,Wog high-dose,and Wog+SIRT1 inhibitor(EX527)groups.After 4 weeks of administration,the volume of ectopic lesions was examined,ELISA was applied to determine the contents of serum estradiol(E2),progestogen(P),interleukin-1β(IL-1β),and IL-6.HE staining was applied to observe pathological damage in ectopic endometrial tissue,Prussian blue staining was applied to detect iron ion aggregation,the colorimetric method was applied to detect the concentration of Fe^(2+),Western blot was applied to detect the protein expression levels of SIRT1,nuclear factor E2 related factor 2(Nrf2),Glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)and ferritin light chain(FTL).Results Compared with the CT group,the ectopic endometrial tissue of rats in EM group had fewer glands,accompanied by a large amount of inflammatory cell infiltration and iron ion deposition,the volume of ectopic lesions and the levels of E2,P,IL-1β,IL-6,and Fe^(2+)increased,the expression levels of SIRT1,Nrf2,GPX4,FTL,and SLC7A11 proteins decreased(P<0.05);compared with the EM group,the ectopic endometrial tissue glands in the Wog L and Wog H groups increased,ion deposition was rare,the volume of ectopic lesions and the levels of E2,P,IL-1β,IL-6,and Fe^(2+)levels decreased,the expression levels of SIRT1,Nrf2,GPX4,FTL,and SLC7A11 proteins increased(P<0.05);compared with the Wog H group,the inflammatory cell infiltration and iron ion deposition in ectopic endometrial tissue of rats in Wog+EX527 group increased,the volume of ectopic lesions and the levels of E2,P,IL-1β,IL-6,and Fe^(2+)increased,the expression levels of SIRT1,Nrf2,GPX4,FTL,and SLC7A11 proteins decreased(P<0.05).Conclusion Wog may inhibit ferroptosis in EM rats by activating SIRT1/Nrf2 signaling pathway.