常山酮通过巨噬细胞介导的子宫内膜基质细胞生物学行为参与调控子宫内膜异位症的作用研究  

Effect of Halofuginone on Proliferation and Apoptosis of Endometrial Stromal Cells through Macrophage-Mediated Pathways in the Pathogenesis of Endometriosis

作　　者：曹文超 CAO Wenchao(Department of Anesthesiology,Beijing Obstetrics and Gynecology Hospital,Capital Medical University,Beijing,100026,China)

机构地区：[1]首都医科大学附属北京妇产医院麻醉科,北京市100026

出　　处：《医学分子生物学杂志》2025年第2期146-152,172,共8页Journal of Medical Molecular Biology

基　　金：首都临床诊疗技术研究及转换应用(省级)(No.Z2011000552085)。

摘　　要：目的探究常山酮(halofuginone,HF)通过调控过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptor gamma,PPARG)-白血病抑制因子(LIF interleukin 6 family cytokine,LIF)轴影响巨噬细胞极化进而调控子宫内膜异位症(endometriosis,EMs)基质细胞生物学行为的分子机制。方法不同浓度的HF干预正常子宫内膜基质细胞(normal endometrial stromal cells,NESCs)与异位子宫内膜基质细胞(ectopic endometrial stromal cells,EESCs),CCK8检测细胞增殖活力,TUNEL检测细胞凋亡。生物信息学分析HF、EMs、巨噬细胞靶点的交集。构建巨噬细胞与EESCs的共培养系统,分为M0+EESCs组、M2+EESCs组、M2+EESCs^(HF)组、M0+EESCs^(oe-PPARG)组、M0+EESCs^(sh-PPARG)组、M0+EESCs^(oe-PPARG+oe-LIF)组、M0+EESCs^(oe-PPARG+HF)组。ELISA检测细胞上清液中M1型巨噬细胞标志物(iNOS、IL-6)与M2型巨噬细胞标志物(Arg-1、TGF-β);CCK8检测和TUNEL分别检测以上EESCs的增殖和凋亡。结果HF对NESCs增殖活力无明显影响(F=0.195,P=0.959),但能够明显抑制EESCs增殖,并促进细胞凋亡(P<0.05)。与M0+EESCs组相比,M2+EESCs组细胞上清液中iNOS、IL-6浓度降低,Arg-1、TGF-β浓度升高,并且EESCs增殖活力增强、凋亡减少,而HF干预则逆转了上述结果(P<0.05)。通过生物信息学分析及文献检索选取PPARG作为本研究关键靶点。与M0+EESCs组相比,M0+EESCs^(oe-PPARG)组细胞上清液中iNOS、IL-6浓度降低,Arg-1、TGF-β浓度升高,EESCs增殖活力增强、凋亡减少(P<0.05);而M0+EESCs^(sh-PPARG)组得到相反的结果(P<0.05)。此外,与M0+EESCs^(oe-PPARG)组相比,上调LIF表达或者HF干预后细胞上清液中iNOS、IL-6浓度升高,Arg-1、TGF-β浓度降低,EESCs增殖活力下降、凋亡增加(P<0.05)。结论HF通过调控PPARG-LIF信号通路从而抑制巨噬细胞M2型极化,抑制EESCs细胞增殖并促进细胞凋亡。Objective To investigate the effect of halofuginone(HF)on proliferation and apoptosis of endometriotic stromal cells(ESCs)in endometriosis(EMs)by regulation of macrophage polarization through peroxisome proliferator-activated receptor gamma(PPARG)-leukemia inhibitory factor(LIF)axis.Methods Normal endometrial stromal cells(NESCs)and ectopic endometrial stromal cells(EESCs)were treated with different concentrations of HF.CCK8 assay was used to detect the cell proliferation activity,and TUNEL assay was used to detect the cell apoptosis.Bioinformatics analysis was performed to identify the intersection of targets of HF,EMs,and macrophage.A co-culture system of macrophages and EESCs was established,cells were then divided into 7 groups:M0+EESCs group、M2+EESCs group、M2+EESCs^(HF) group、M0+EESCs^(oe-PPARG)group、M0+EESCs^(sh-PPARG)group、M0+EESCs^(oe-PPARG)+oe-LIF group、M0+EESCs^(oe-PPARG)+HF group.ELISA was used to detect the concentrations of M1 macrophage markers(iNOS,IL-6)and M2 macrophage markers(Arg-1,TGF-β)in the cell supernatant.CCK8 and TUNEL assay were used to detect the proliferation and apoptosis of EESCs,respectively.Results HF had no significant effect on the proliferation activity of NESCs(F=0.195,P=0.959),but significantly inhibited the proliferation of EESCs and promoted cell apoptosis(P<0.05).The levels of iNOS and IL-6 in the M2+EESCs group were decreased when compared with those in the M0+EESCs group,while the levels of Arg-1 and TGF-βwere increased.In addition,the proliferation activity of EESCs was increased and the apoptosis was decreased.HF intervention reversed the results above(P<0.05).PPARG was identified as a key target in this study through bioinformatics analysis and literature search.The levels of iNOS and IL-6 in the cell supernatant of the M0+EESCs^(oe-PPARG)group were decreased,while the levels of Arg-1 and TGF-βwere increased when compared with those in the M0+EESCs group.The proliferation activity of EESCs were increased and apoptosis were decreased(P<0.05),while an oppo

关 键 词：常山酮 过氧化物酶体增殖物激活受体Γ 白血病抑制因子 巨噬细胞 子宫内膜异位症 子宫内膜异位基质细胞 

分 类 号：R711.71[医药卫生—妇产科学]