雷公藤多甙抑制EM患者腹腔巨噬细胞及相应细胞模型分泌TNF-α的实验研究

Experimental study of Tripterygium wilfordii Hook.f.for inhibiting TNF-αsecretion in peritoneal macrophage of endometriosis patients and relevant cell model

目的观察雷公藤多甙(TWP)对子宫内膜异位症(EM)患者腹腔巨噬细胞和小鼠腹腔活化巨噬细胞释放TNF-α的影响,以探讨TWP用于临床治疗EM的机制并建立细胞模型,为后续研究做准备。方法收集16例经腹腔镜检查确诊为EM患者腹腔液分离纯化巨噬细胞,并给予不同剂量的TWP进行共同孵育;同时分离纯化小鼠腹腔巨噬细胞,用脂多糖(LPS)激活以获得细胞模型,同样予TWP进行共同孵育。分别以间接MTT法和RT-PCR法在蛋白质水平和基因水平检测该药物影响激活状态巨噬细胞分泌TNF-α含量的变化。结果 EM患者腹腔液中TNF-α含量显著高于非EM妇女,在4～24 h后TWP(6.25～100μg/L)显著抑制了TNF-α的分泌。结论 TWP在蛋白质和基因水平均能有效抑制激活巨噬细胞分泌TNF-α。

Objective It is to observe the effect of Tripterygium wilfordii Hook. f. （TWP） on the activity of TNF-α which excreted from peritoneal elicited macrophage of patients with endometriosis（EM） and peritoneal elicited macrophage activated by LPS in mice, so to provide new experimental proofs and academic bases for clinical therapy on EM and get ready for next study. Methods The peritoneal macrophage of mice and 16 cases of patients with EM diagnosed by laparoscopes examination were separated and purified, then they were cultured in vitro with a series concentration of TWP during different time. The content changes of TNF -α at protein level and gene level were detected through MTT colorimetry and RT - PCR respectively. Results The level of TNF -α in peritoneal fluid of patients with EM was significantly higher than that of women without EM. The level of TNF-α secreted by M φeau be inhibited significantly（ P 〈 0.01 ） by TWP （6.25 -100μg/L） during 4 -24 h. Conclusion TWP can effectively inhibit TNF - α secretion at protein and gene level in Mq〉φ.