Zhang Zhenzhen, Cheng Xiaolan, Gui Tao, Tao Jia, Huang Meihua, Zhu Li, Luo Mei, Cao Peng, Wan Guiping
Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing 210028, Jiangsu, PR China; Jiangsu Province Academy of Traditional Chinese Medicine, Nanjing 210028, Jiangsu, PR China.
J Ethnopharmacol. 2016 Dec 24;194:386-394. doi: 10.1016/j.jep.2016.07.019. Epub 2016 Jul 9.
Wenshen Xiaozheng Tang (WXT), a traditional Chinese medicine prescription, exerted a good therapeutic effect on endometriosis. However, the underlying mechanism is unclear. In the present study, we sought to evaluate the effect of WXT on the proliferation and migration of ectopic endometriotic stromal cells and explore the potential molecular mechanism.
Primary stromal cells derived from ectopic endometriotic lesions of patients with endometriosis were isolated and cultured. The inhibition effect of WXT on cell proliferation was determined by MTT. Apoptosis of ectopic endometriotic cells treated with WXT was analyzed with Annexin V-FITC/7-AAD staining. The activation of caspases was detected by western blot analysis. The influence of WXT on migration of ectopic endometriotic cells was measured by scratch wound healing assay and Transwell assay. The DNA binding activity of NF-κB and the expression of nuclear p65 protein were determined by electrophoretic mobility shift assay and western blot analysis, respectively. The impact of WXT on the expression of NF-κB regulated gene products involved in apoptosis and migration was determined by western blot analysis.
WXT inhibited the proliferation of ectopic endometriotic cells in a time- and dose-dependent manner. In addition, WXT treatment resulted in significant induction of apoptosis through the activation of caspases and inhibition of migration in ectopic endometriotic cells. WXT notably suppressed constitutive NF-κB-DNA-binding activity as well as TNF-α induced nuclear translocation of NF-κB p65 subunit in ectopic endometriotic cells. Moreover, WXT diminished the expression of NF-κB regulated gene products involved in apoptosis and migration, including c-IAP1, c-IAP2, XIAP, survivin, Mcl-1, COX-2 and MMP-9.
Our results indicate that WXT induces apoptosis and inhibits migration of ectopic endometriotic stromal cells.
温肾消症汤（WXT）是一种中药方剂，对子宫内膜异位症具有良好的治疗效果。然而，其潜在机制尚不清楚。在本研究中，我们旨在评估WXT对异位子宫内膜间质细胞增殖和迁移的影响，并探索其潜在的分子机制。
分离并培养来自子宫内膜异位症患者异位内膜病变的原代间质细胞。通过MTT法测定WXT对细胞增殖的抑制作用。用Annexin V-FITC/7-AAD染色分析WXT处理的异位子宫内膜细胞的凋亡情况。通过蛋白质免疫印迹分析检测半胱天冬酶的激活情况。通过划痕伤口愈合试验和Transwell试验测定WXT对异位子宫内膜细胞迁移的影响。分别通过电泳迁移率变动分析和蛋白质免疫印迹分析测定NF-κB的DNA结合活性和核p65蛋白的表达。通过蛋白质免疫印迹分析确定WXT对参与凋亡和迁移的NF-κB调节基因产物表达的影响。
WXT以时间和剂量依赖性方式抑制异位子宫内膜细胞的增殖。此外，WXT处理通过激活半胱天冬酶和抑制异位子宫内膜细胞的迁移导致显著的凋亡诱导。WXT显著抑制异位子宫内膜细胞中组成型NF-κB-DNA结合活性以及TNF-α诱导的NF-κB p65亚基的核转位。此外，WXT降低了参与凋亡和迁移的NF-κB调节基因产物的表达，包括c-IAP1、c-IAP2、XIAP、survivin、Mcl-1、COX-2和MMP-9。
我们的结果表明，WXT诱导异位子宫内膜间质细胞凋亡并抑制其迁移。
