Buathong Norathee, Poonyachoti Sutthasinee, Deachapunya Chatsri
J Med Assoc Thai. 2016 Nov;99 Suppl 8:S134-S141.
The present study aimed to investigate the anti-inflammatory effect of phytoestrogen genistein (Ge) on the secretion of interleukin 6 (IL6) under lipopolysaccharide (LPS) stimulated conditions in human endometrial epithelial cell line RL95-2. The effects of Ge on expression of TLRs 2, 3, 4 and 9 proteins in response to the inflammatory development induced by LPS were also examined and compared with those of 17β-estradiol (E2).
The RL95-2 cells were cultured in the estrogen-deprived media with or without bacterial endotoxin LPS 30 min prior to incubation with Ge (10-7, 10-6 or 10-5 M) or E2 (10-9 M) for 48 h. The culture media were collected at 1 and 24 h for IL6 measurement by the enzyme linked immunosorbent assay and the cell lysate for TLR protein expression analyzed by semi-quantitative Western blot.
Ge or E2 did not alter the IL6 level in the absence of LPS; however, treatment with either Ge or E2 for 1 h up to 24 h decreased the IL6 level stimulated by LPS. The cells challenged with LPS significantly upregulated the TLRs 2 and 9 but suppressed the TLRs 3 and 4 protein expression. Forty eight h-treatment with Ge had no effect on the TLRs expression, whereas E2 down-regulated the increased TLR9 induced by LPS.
Ge suppressed the inflammatory response by decreasing the IL6 level, but not associated with the alteration of TLRs-mediated pathway inducedby bacterial infection. In contrast, E2 decreased the IL6 secretion possibly due to the opposition on LPS-induced TLR9 expression. This provides the potential evidence of soy isoflavone genistein in alleviating the inflammation of endometrium following bacterial invasion.
本研究旨在探讨植物雌激素染料木黄酮（Ge）在脂多糖（LPS）刺激条件下对人子宫内膜上皮细胞系RL95 - 2白细胞介素6（IL6）分泌的抗炎作用。还检测了Ge对LPS诱导的炎症反应中Toll样受体（TLRs）2、3、4和9蛋白表达的影响，并与17β - 雌二醇（E2）进行比较。
将RL95 - 2细胞在缺乏雌激素的培养基中培养，在与Ge（10 - 7、10 - 6或10 - 5 M）或E2（10 - 9 M）孵育48小时前30分钟，加入或不加入细菌内毒素LPS。在1小时和24小时收集培养基用于通过酶联免疫吸附测定法测量IL6，并收集细胞裂解物用于通过半定量蛋白质印迹法分析TLR蛋白表达。
在不存在LPS的情况下，Ge或E2不改变IL6水平；然而，用Ge或E2处理1小时至24小时可降低LPS刺激的IL6水平。用LPS刺激的细胞显著上调TLRs 2和9，但抑制TLRs 3和4蛋白表达。用Ge处理48小时对TLRs表达无影响，而E2下调LPS诱导的TLR9增加。
Ge通过降低IL6水平抑制炎症反应，但与细菌感染诱导的TLRs介导途径的改变无关。相比之下，E2可能由于对LPS诱导的TLR9表达的拮抗作用而降低IL6分泌。这为大豆异黄酮染料木黄酮减轻细菌入侵后子宫内膜炎症提供了潜在证据。
