川芎嗪抑制TLR4/MyD88/NF-κB信号通路减轻子宫内膜异位症大鼠炎症反应

Ligustrazine mitigates inflammatory response in endometriosis rats by inhibiting TLR4/MyD88/NF-κB signaling pathway

目的探讨川芎嗪抑制Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核因子κB(NF-κB)信号通路减轻子宫内膜异位症(EMs)大鼠炎症反应的作用。方法自体子宫移植法构建EMs大鼠模型,随机分为EMs组、川芎嗪(低、中、高)剂量组、阳性药物组,每组各12只;进行假手术的12只大鼠作为假手术组。游标卡尺测定异位病灶体积,HE染色检测异位内膜组织病理学改变,ELISA检测血清性激素及炎症因子水平,qRT-PCR、Western blot、免疫荧光染色分别检测异位内膜组织中炎症因子mRNA表达、TLR4/MyD88/NF-κB信号通路相关蛋白表达及NF-κB p65入核情况。结果与假手术组比较,EMs组大鼠异位内膜组织出现明显病理损伤,血清雌二醇(E2)、孕激素(P)含量、肿瘤坏死因子-α(TNF-α)、白介素(IL)-1β、IL-6水平及异位内膜组织中TNF-α、IL-1β、IL-6 mRNA、总蛋白TLR4、MyD88、p-NF-κB p65和核蛋白NF-κB p65表达水平升高,血清促卵泡激素(FSH)、促黄体生成素(LH)含量降低(P<0.05),同时NF-κB p65蛋白大量入核;与EMs组比较,川芎嗪(低、中、高)剂量组和阳性药物组可缓解EMs模型大鼠的上述改变并降低异位病灶体积,且川芎嗪高剂量组和阳性药物组对EMs大鼠的作用效果较好且相近。结论川芎嗪可能通过抑制TLR4/MyD88/NF-κB信号通路激活减轻EMs大鼠炎症反应。

To investigate the mitigation effect of ligustrazine on the inflammatory response of endometriosis(EMs)rats by inhibiting Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factorκB(NF-κB)signaling pathway.The EMs rat model was established by autologous uterine transplantation,and they were randomly divided into EMs group,ligustrazine(low,medium,and high doses)groups and positive drug group,with 12 rats in each group;12 rats subjected to sham operation were regarded as the sham operation group.Vernier caliper was used to measure the volume of ectopic lesions;HE staining was used to detect the histopathological changes of ectopic endometrium;ELISA was used to detect the levels of serum sex hormones and inflammatory factors;qRT-PCR,Western blot and immunofluorescence staining were used to detect the mRNA expression levels of inflammatory factors,the expression levels of TLR4/MyD88/NF-κB signaling pathway-related proteins and the nuclear entry of NF-κB p65 in ectopic endometrial tissue,respectively.Compared with the sham operation group,the EMs group showed obvious pathological damage in ectopic endometrium,higher levels of estradiol(E2),progesterone(P),tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6 in serum,and higher expression levels of TNF-α,IL-1βand IL-6 mRNA,as well as higher expression levels of total protein TLR4,MyD88,p-NF-κB p65 and nuclear protein NF-κB p65 in ectopic endometrial tissue,but lower levels of folliclestimulating hormone(FSH)and luteinizing hormone(LH)in serum decreased(P<0.05).Furthermore,in the EMs group,a large number of NF-κB p65 proteins entered the nucleus.Compared with EMs group,ligustrazine(low,medium and high doses)groups and positive drug group can alleviate the above changes and reduce the volume of ectopic lesions of EMs model rats,and the effects of ligustrazine high dose group and positive drug group were well and similar.In conclusion,ligustrazine may alleviate the inflammatory response of EMs rats by inhibiting the activation of TLR4/MyD88/NF-κB signal pathway.