姜黄素对子宫内膜异位症大鼠PI3K/AKT/GSK-3β通路及肥大细胞活化的影响

Effects of curcumin on PI3K/AKT/GSK-3β pathway and mast cell activation in endometriosis rats

目的:观察姜黄素对子宫内膜异位症(EM)大鼠的影响,并初步探讨其作用机制。方法:将SD大鼠随机分为假手术组、模型组、姜黄素低、中、高剂量组和阳性对照组,除假手术组外均通过外科手术将右侧自体子宫组织移植到大鼠左侧腹部腹壁,缝合腹部建立EM模型,假手术组仅分离右侧子宫内膜,不进行自体移植。假手术组和模型组给予等量蒸馏水灌胃;姜黄素低、中、高剂量组分别给予60、120、240 mg/kg姜黄素灌胃;阳性对照组给予125 mg/kg孕三烯酮灌胃,1次/d,连续灌胃21 d,解剖后观察各组大鼠内异部位内异灶和盆腔黏连情况;HE染色观察各组大鼠内移灶组织病理学变化;甲苯胺蓝染色观察各组大鼠内异灶组织中肥大细胞总数量及脱落颗粒肥大细胞数量;采用TNF-α、IL-1β和IL-6 ELISA试剂盒检测各炎症因子水平;Western blot检测各组大鼠内异灶组织中磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)和糖原合成酶激酶3β(GSK-3β)蛋白表达。结果:假手术组大鼠子宫内膜完整,上皮细胞柱状排列整齐,腺体自然;模型组大鼠异位内膜组织生长状态良好,腺体上皮细胞和基质细胞均正常;姜黄素低、中、高剂量组大鼠异位子宫内膜上皮细胞形状逐渐变为单层柱状,腺体和基质细胞数量逐渐减少,并出现空化现象;阳性对照组上述现象介于姜黄素中剂量组和高剂量组之间。与假手术组相比,模型组子宫内膜异位症大鼠内异灶面积、盆腔黏连评分、内异灶组织肥大细胞总数、脱颗粒肥大细胞数和脱颗粒肥大细胞/肥大细胞总数、TNF-α、IL-1β和IL-6水平、PI3K、p-AKT和p-GSK-3β蛋白表达均显著升高(P<0.05);与模型组相比,姜黄素低、中、高剂量组子宫内膜异位症大鼠内异灶面积和盆腔黏连评分、异位灶组织脱颗粒肥大细胞数和脱颗粒肥大细胞/肥大细胞总数、TNF-α、IL-1β和IL-6水平、PI3K、p-AKT和p-GSK-3β蛋白表达均降低(P<0.05),阳性对照组大鼠上述指标均显著降低(P<0.05)。结论:姜黄素可阻抑EM大鼠内异灶组织生长,可能与抑制PI3K/AKT/GSK3β通路、降低肥大细胞活化及炎症反应相关。

Objective:To observe the effect of curcumin on endometriosis(EM)rats and explore its mechanism.Methods:SDrats were randomly divided into sham operation group,model group,low,medium,high dose curcumin groups and positive controlgroup.Except sham operation group,right autogenous uterine tissue was transplanted into the left abdominal wall of rats by surgery,and sutured the abdomen to establish EM model,while in sham operation group,only the right endometrium was isolated without autol-ogous transplantation.Sham operation group and model group were given the same amount of distilled water by gavage;the low,medi-um and high dose curcumin groups were given 60,120 and 240 mg/kg curcumin respectively;positive control group was given 125 mg/kggestrinone by gavage,once a day for 21 days,after dissection,the heterotopic focus and pelvic adhesion of heterotopic part wereobserved;HE staining was used to observe histopathological changes of the transplanted foci;toluidine blue staining was used to ob-serve the total number of mast cells and number of mast cells in exfoliated granules of rats in each group;TNF-α,IL-1βand IL-6 ELI-SA kits were used to detect levels of inflammatory factors;Western blot was used to detect protein expressions of phosphatidylinositol3-kinase(PI3 K),protein kinase B(AKT)and glycogen synthase kinase-3β(GSK-3β).Results:In sham operation group,the endo-metrium was intact,the epithelial cells were arranged orderly and the glands were natural;in model group,the growth of ectopic endo-metrium was good,and the glandular epithelial cells and stromal cells were normal;in low,medium and high dose curcumin groups,shape of ectopic endometrial epithelial cells gradually changed into monolayer columnar shape,number of glands and stromal cells de-creased gradually,and cavitation phenomenon appeared;the above phenomena in positive control group were between those in medi-um dose curcumin group and high dose curcumin group.Compared with those in sham operation group,the area of heterotopic focus,pelvic adhesion score,total number of mast cells,number of degranulated mast cells and total number of degranulated mast cells/mast cells,levels of TNF-α,IL-1βand IL-6,protein expression levels of PI3 K,p-AKT and p-GSK-3βwere significantly higher in model group(P<0.05);compared with those in model group,the area of heterotopic focus,pelvic adhesion score,number of degranu-lated mast cells and the total number of degranulated mast cells/mast cells,levels of TNF-α,IL-1βand IL-6 protein expression levelsof PI3 K,p-AKT and p-GSK-3βwere lower in low,medium and high dose curcumin groups(P<0.05),the above indexes in positivecontrol group were significantly lower(P<0.05).Conclusion:Curcumin can inhibit the growth of ectopic tissue in EM rats,which maybe related to the inhibition of PI3 K/AKT/GSK-3βpathway and the decreases of mast cell activation and inflammatory reaction.