黄芪多糖对孕早期蜕膜TLR4表达与功能的影响

EFFECTS OF ASTRAGALAN ON EXPRESSION AND FUNCTION OF TLR4 IN DECIDUAL CELLS OF EARLY PREGNANCY

目的:观察黄芪多糖通过孕早期蜕膜TLR4途径对IL-6、TNF-α分泌的影响。方法:采用流式细胞术检测孕早期蜕膜TLR4的表达,并对孕早期蜕膜进行原代培养,将其分为两大组,PBS对照组、LPS刺激组,用0、25、50、125、250mg/L黄芪多糖预处理两组蜕膜细胞48小时,再分别用PBS、LPS处理24小时,采用酶联免疫吸附试验方法检测上清液TNF-α、IL-6的分泌,用流式细胞仪检测蜕膜细胞TLR4的表达。结果:孕早期蜕膜功能性表达TLR4,用LPS刺激细胞后,TLR4表达上调,IL-6、TNF-α的分泌显著增加,用25、50、125、250mg/L黄芪多糖预处理蜕膜细胞48小时后,250mg/L黄芪多糖可以抑制TLR4表达的上调,同时可以抑制LPS刺激蜕膜细胞引起的TNF-α分泌增加,而25、50、125mg/L黄芪多糖不能抑制LPS刺激引起的TLR4表达的上调;但25、50、125、250mg/L黄芪多糖均可以引起IL-6分泌的增加。未经LPS刺激,用25、50、125、250mg/L黄芪多糖作用蜕膜细胞,蜕膜TLR4的表达及IL-6、TNF-α的分泌则无显著性变化。结论:TLR功能性表达于孕早期蜕膜;黄芪多糖可以通过改变蜕膜TLR介导的免疫功能影响母胎界面免疫微环境。

Objective:To observe the effects of astragalan on secretion of IL-6 and TNF-α of decidual cells mediated by toll like receptor(TLR4).Methods:The expression of TLRs in human decidual cells in early pregnancy was detected by flow cytometry,decidual cells were primarily cultured and divided into two groups,one was cultured with PBS to be blank,the other was stimulated with LPS to be model group.Decidual cells of the two group were cultured with different concentration astragalan(0,25,50,125,250mg/L) for 48 hours,than respectively added PBS and LPS to cultured cells for 24 hours,supernatant fluid was collected to determine the content of IL-6 and TNG-α by ELISA,the expression of TLR4 was detect by flow cytometry。 Results:TLR4 was expressed functionally in primary human decidual cells.The expression of TLR4 elevated after stimulated by LPS,the secretion of IL-6 and TNF-α elevated obviously,250mg/L astragalan could inhibit the increasing of TLR4 expression and inhibit the raising of TNF-α secretion caused by LPS,the other concentration astragalan couldn't inhibit the up-regulation of TLR4 expression,but 25,50,125,250mg/L astragalan could cause the increasing of IL-6 secretion.Astragalan of each concentration couldn't significantly influence the TLR4 expression and IL-6,TNF-α secretion of human decidual cells without stimulated with LPS.Conclusion:Toll like receptor was expressed functionally in primary human decidual cells.Astragalan can effect the maternal-fetal interface microenviroment by regulating the immune function induce by TLR in decidual cells in early pregnancy.