Wieser Fritz, Yu Jie, Park John, Gaeddert Andrew, Cohen Misha, Vigne Jean-Louis, Taylor Robert N
Department of Gynecology and Obstetrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Biol Reprod. 2009 Aug;81(2):371-7. doi: 10.1095/biolreprod.108.075069. Epub 2009 Apr 29.
Growing evidence suggests that medicinal herbs have direct actions on endometrial cells. By screening multiple herbs using an in vitro model of endometriosis, we found that a commonly used herbal formula exerted considerable antiproliferative effects. Our purpose was to investigate the effects of this antiendometriosis herbal mixture on cell proliferation, apoptosis, and CCL5 expression and secretion in endometriotic stromal cells in vitro. Isolated normal endometrial, eutopic, and ectopic endometriotic stromal cells were cultured under established conditions. Cell proliferation, apoptosis, and CCL5 gene expression protein secretion was evaluated after incubation with different concentrations of an antiendometriosis herbal mixture extract. Cell proliferation was assessed by cell counting, (3)H-thymidine incorporation, and MTS assays. Apoptosis was determined by blotting using anti-cleaved caspase 3 antibodies and by a TUNEL assay. CCL5 gene expression and protein secretion were determined by transient transfection of gene promoter reporters and ELISAs in cell supernatants. Extracts of a traditional herbal mixture dose-dependently decreased cell proliferation in normal, eutopic, and ectopic endometriotic stromal cells. (3)H-Thymidine uptake and MTS confirmed these findings. The herbal extracts induced apoptosis, as evidenced by activation of caspase 3 and the presence of TUNEL-positive cells after treatment. The herbal extracts also suppressed CCL5 gene transcription and protein secretion in endometriotic stromal cells, even when corrected for cell number. Extracts from a medicinal herbal mixture have direct effects on cell proliferation, apoptosis, and CCL5 production in endometriotic stromal cells. Our findings support the further investigation of novel, potentially safe and well-tolerated botanical products as future endometriosis treatments.
越来越多的证据表明,草药对子宫内膜细胞有直接作用。通过使用子宫内膜异位症的体外模型筛选多种草药,我们发现一种常用的草药配方具有相当大的抗增殖作用。我们的目的是研究这种抗子宫内膜异位症草药混合物对体外子宫内膜异位间质细胞的细胞增殖、凋亡和CCL5表达及分泌的影响。分离出的正常子宫内膜、在位和异位子宫内膜间质细胞在既定条件下培养。用不同浓度的抗子宫内膜异位症草药混合物提取物孵育后,评估细胞增殖、凋亡和CCL5基因表达蛋白分泌。通过细胞计数、³H-胸腺嘧啶核苷掺入和MTS测定评估细胞增殖。通过使用抗切割的半胱天冬酶3抗体进行印迹和TUNEL测定来确定凋亡。通过基因启动子报告基因的瞬时转染和细胞上清液中的ELISA来确定CCL5基因表达和蛋白分泌。传统草药混合物的提取物剂量依赖性地降低正常、在位和异位子宫内膜间质细胞的细胞增殖。³H-胸腺嘧啶核苷摄取和MTS证实了这些发现。草药提取物诱导凋亡,治疗后半胱天冬酶3的激活和TUNEL阳性细胞的存在证明了这一点。即使校正细胞数量后,草药提取物也抑制子宫内膜异位间质细胞中CCL5基因转录和蛋白分泌。药用草药混合物的提取物对子宫内膜异位间质细胞的细胞增殖、凋亡和CCL5产生有直接影响。我们的发现支持进一步研究新型、可能安全且耐受性良好的植物产品作为未来子宫内膜异位症的治疗方法。