腹腔液中巨噬细胞产生血管内皮细胞生长因子及其受体的表达

目的 :探讨腹腔环境在子宫内膜异位症发病机制中的作用 ,以及腹腔液血管内皮细胞生长因子 (VEGF)的来源和调节。方法 :将 14例子宫内膜异位症患者和 10例正常妇女腹腔液中的巨噬细胞体外培养 ,并在培养的正常妇女腹腔液巨噬细胞中加入 17 β雌二醇、孕酮和脂多糖 (LPS)共培养。用ELISA法检测培养的巨噬细胞上清液中VEGF的水平。同时将二者无血清培养的上清液 ,加入到内皮细胞中培养 ,用MTT比色法检测其对内皮细胞增殖活性的影响。用免疫组化法检测两者的巨噬细胞表达受体flt 1和flt 4的水平。结果 :子宫内膜异位症患者的巨噬细胞培养上清液中 ,VEGF的浓度明显高于正常对照组 (P <0 .0 5 ) ,且无周期性变化 (P >0 .0 5 )。子宫内膜异位症患者的巨噬细胞无血清培养上清液 ,在体外能显著提高内皮细胞的增殖活性 (P <0 .0 5 )。雌激素和孕激素能显著增加体外巨噬细胞分泌VEGF的水平 (P <0 .0 5 ) ,与LPS组相比较差异显著 (P <0 .0 5 ) ,即激素的调节作用大于LPS的作用。但雌激素和孕激素调节巨噬细胞分泌VEGF的程度无明显差别 (P >0 .0 5 )。LPS激活的巨噬细胞能增加雌激素、孕激素对巨噬细胞分泌VEGF的调节作用 (P <0 .0 5 )。巨噬细胞能表达受体flt 1和flt 4 ,无周期性变化 (P >0 .0 5 )。结论 :子宫内膜

VEGF production and its receptor expression by MΦs in peritoneal fluid

AIM: To explore the role of microenvironment of peritoneal fluid in pathogenesis of endometriosis, the source and its regulation in peritoneal fluid. METHODS: Peritoneal Mphis from 14 patients with endometriosis (endometriosis group)and 10 normal women ( control group) were cultured in vitro. Either 17-l estradiol, progesterone and LPS or their combinant were added to cultured peritoneal Mos from control group. VEGF levels in culture supernatant of M(Ds from two groups were detected by ELISA. Culture supernatant of serum-free media in two groups were added to the cultured endothelial cells. Effect of the supernatant on endothelial cell proliferation activity was examined by MTT colorimetry at wavelength of 450 nm. The expressions of Fit-1 and Flt-4 on the Mos from two groups were determined by immunohistochemical staining. RESULTS: The VEGF level in culture supernatant of Mps from endometriosis group was significantly higher than that from control group (P < 0. 05 ), and had no cyclic variation (P > 0. 05). As compared with LPS, estrogen and progentrogen could elevate notably the level ofVEGF secreted by Ms ( P < 0. 05 ). However, the regulation of MWs secreted VEGF level by estrogen and progestoron had no marked difference. The Mos activated by LPS could strengthen the regulation of VEGF secreted by Mphis under estrogen and progestron stimulation. Culture supernatant of serum-free media for Mos could promote significantly the proliferation of endothelial cells ( P < 0. 05 ). Fit-land Flt-4 on Mos was expressed without cyclic variation (P>0.05). CONCLUSION: The VEGF may be secreted by Mos in peritoneal fluid of endometriosis group. The endothelial cell proliferation and angiogenesis are enhanced by change of microenvironment of peritoneal fluid.