TLR4信号介导内膜间质细胞侵袭增殖促子宫腺肌症发病的研究

TLR4-mediated stromal cells acquire an invasive phenotype and implicated in the pathogenesis of adenomyosis

目的:探讨Toll样受体4(TLR4)信号活化内膜间质在腺肌症发生与发展中的调控作用。方法:Western blot法检测腺肌病在位子宫内膜及病灶内膜中TLR4表达情况;建立原代间质体系,lipopolysaccharide(LPS)刺激靶细胞,CCK-8技术观察细胞增殖活力;Western blot法检测TLR4核心信号表达;Western blot法和酶联免疫吸附测定(ELISA)技术检测和分析间质细胞炎性增殖及侵袭生长相关因子表达;采用TLR4拮抗剂VIPER处理细胞,验证TLR4信号对间质生物效应的影响。结果:TLR4表达于细胞质膜,在腺肌症病灶内膜中表达最高;LPS刺激使间质细胞活力增强、TLR4核心信号及增殖侵袭因子表达增加;VIPER可抑制上述因子表达。结论:LPS/TLR4信号活促进内膜间质细胞炎性增殖、侵袭生长,在子宫腺肌症发病中发挥重要作用,提示宫腔感染可能与腺肌病发病具有一定的相关性。

Objective： To investigate that the TLR4 signal pathway-induced cell biologic effects was associated with the development and progress of adenomyosis,which involved in the pathogenesis of adenomyosis. Method： The expression of TLR4 in the endometrium with adenomyosis was detected by WB. In vitro stromal cells models were established,and the endometrial stromal cells were treated with LPS and TLR4 antagonist. The cells viability was tested through CCK8,the key protein of TLR4 signal pathway（ TLR4,MD2,My D88,NF-κB） were detected by WB,moreover,the inflammatory proliferation and invasive growth of experimental cells were observed and analyzed by WB / ELISA. Result： The expression of TLR4 was significantly higher in eutopic / ectopic endometrium with adenomyosis than that in the control groups showed in WB.The viability of experimental cells treated with LPS was showed significantly enhancer than the non-treatedand that could be inhibited with TLR4 antagonist,VIPER. The expressions of TLR4,MD2,My D88,NF-κB and cellular inflammatory proliferation and invasive growth factors（ VEGF,EGF,IL-6,TGF-β,MMP2） were observed increased significantly（ E-cadherin decreased） by LPS-treated in stromal cells of all groups were induced intensely by LPS,and the cellular effect could be blocked by VIPER. Conclusion： Our results showed that stromal cells were activated by TLR4 signal pathway,which processed the cellular inflammatory proliferation and invasive growth. The above results might reveal that TLR4 signal pathway involved in the pathogenesis of adenomyosis,which might be associated with the patients who experienced intrauterine microbes.