黄芪甲苷对子宫内膜异位症大鼠TLR4-MyD88信号通路的影响

Effect of astragaloside A on TLR4-MyD88 signal pathway in rats with endometriosis

目的分析黄芪甲苷对子宫内膜异位症大鼠的作用及对Toll样受体4(TLR4)-髓样分化因子(MyD88)信号通路的影响。方法将50只SPF级无交配史SD雌性大鼠按随机数字表法分为模型组、对照组和黄芪甲苷低、中、高剂量组,每组10只,除对照组外均构建子宫内膜异位症模型。黄芪甲苷低、中、高剂量组分别给予20、40、80 mg/kg黄芪甲苷灌胃处理,对照组、模型组灌胃等量生理盐水,连续给药28 d。测量各组大鼠异位子宫内膜体积;测定血清性激素指标雌二醇(E_(2))、孕酮(P)、促黄体生成激素(LH)、卵泡刺激素(FSH),以主炎症因子指标肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-8水平;苏木精-伊红染色观察子宫内膜组织病理变化;蛋白质印迹法(Western blotting)检测子宫内膜异位组织中TLR4-MyD88信号通路蛋白表达。结果与对照组相比,模型组大鼠异位子宫内膜体积增大(P<0.05),以及血清E_(2)、P、LH、TNF-α、IL-6、IL-8水平升高(P<0.05);与对照组相比,模型组大鼠移植内膜生长,被覆柱状上皮,间质较多,可见炎性浸润,子宫内膜组织TLR4、MyD88、p-NF-κB、NF-κB蛋白水平升高(P<0.05)。与模型组相比,黄芪甲苷低剂量组、中剂量组、高剂量组大鼠异位子宫内膜体积缩小(P<0.05),以及血清E_(2)、P、LH、TNF-α、IL-6、IL-8水平下降(P<0.05),且存在剂量依赖性(P<0.05);与模型组相比,黄芪甲苷低剂量组、中剂量组、高剂量组大鼠移植内膜变少,生长不完整,腺体减少或消失,腺上皮萎缩,子宫内膜组织TLR4、MyD88、p-NF-κB、NF-κB蛋白水平均降低(P<0.05),且存在剂量依赖性(P<0.05)。结论黄芪甲苷可剂量依赖地抑制子宫内膜异位症大鼠异位病灶生成,下调性激素水平,降低炎症反应,抑制TLR4-MyD88信号通路激活。

Objective To analyze the effect of astragaloside A on the rats with endometriosis and its influences on Toll like receptor 4(TLR4)-myeloid differentiation factor(MyD88)signal pathway.Methods Fifty SPF SD female rats with no copulative history were divided into the model group,control group and low,medium and high astragaloside A doses groups according to the random number table method,10 cases in each group.The endometriosis models were constructed except the control group.The astragaloside A low,medium and high doses groups were given 20,40 and 80 mg/kg astragaloside A by gavage,respectively,while the control group and model group were given the same amount of normal saline by gavage,and the medication was continuously administered for 28 d.The ectopic endometrial volume was measured in each group.The levels of serum sex hormones indexes estradiol(E_(2)),progesterone(P),luteinizing hormone(LH),follicle-stimulating hormone(FSH),inflammatory factors indexes,tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-8 were measured.The hematoxylin-eosin staining was used to observe the pathological changes of endometrial tissue.The protein expression of TLR4-MyD88 signaling pathway in endometriosis tissue was detected by Western blotting.Results Compared with the control group,the ectopic endometrial volume and the levels of serum E_(2),P,LH,TNF-α,IL-6 and IL-8 in the model group were increased(P<0.05).Compared with the control group,there were more rat transplanted intimal growth,covering columnar epithelium and more stroma,inflammatory infiltration could be seen,the expressions of TLR4,MyD88,p-NF-κB,NF-κB protein in endometrium tissue of the model group were increased(P<0.05).Compared with the model group,the ectopic endometrial volume and the levels of serum E_(2),P,LH,TNF-α,IL-6,IL-8 in the low dose astragaloside A group,medium dose astragaloside A group and high dose astragaloside A group were decreased(P<0.05),moreover there was a dose dependence(P<0.05).Compared with the model group,the transplanted intima in the low dose astragaloside A group,medium dose astragaloside A group and high dose astragaloside A group was decreased,the growth was incomplete,the glands were decreased or disappeared,and the glandular epithelium had atrophy,the levels of TLR4,MyD88,p-NF-κB,NF-κB protein in endometrium tissue were decreased(P<0.05),moreover there was a dose dependence(P<0.05).Conclusion Astragaloside A could dose-dependently inhibit the formation of ectopic lesions in the rats with endometriosis,down-regulate the sex hormone levels,reduce the inflammatory reaction,and inhibit the activation of TLR4-MyD88 signal pathway.