日本血吸虫不同发育阶段抗原对人早期胚胎滋养细胞的影响

Effects of Schistosoma japonicum antigens during different development stages on first trimester human placental trophoblast cells

目的 探讨日本血吸虫不同发育阶段抗原对原代培养的人早孕期滋养细胞的分泌功能的影响，为疫区育龄妇女及孕产妇的临床寄生虫防治提供科学依据。 方法 收集6-10孕周行人工流产术的正常妊娠妇女的绒毛组织；分离、培养滋养细胞；液态芯片ProcartaPlex法检测给予25 μg/ml虫卵排泌抗原（ex/secretory antigen, ES）、可溶性虫卵抗原（soluble egg antigen, SEA）和可溶性虫体抗原（soluble worm antigen, SWA）18 h后滋养细胞培养上清中细胞因子的分泌水平。 结果 ES可以明显促进人早孕期滋养细胞分泌IL-6和IL-8，其浓度为（155.58±20.07）pg/ml（t=8.472，P〈0.05）和（12.78±2.35）ng/ml（t=8.006，P〈0.05），而SEA仅明显促进滋养细胞分泌IL-8，其浓度为（14.18±2.24）ng/ml（t=7.413，P〈0.05）。SEA可以促进滋养细胞分泌趋化因子单核细胞趋化因子-1（monocyte chemotactic protein 1，MCP-1），其浓度为（212.2±14.57）pg/ml（t=15.640，P均〈0.05）。ES和SWA均可以明显促进滋养细胞分泌转化生长因子-β1（transforming growth factor-β1，TGF-β1），其浓度为（78.56±5.71）pg/ml和（66.61±5.55）pg/ml（t=8.892，6.538，P均〈0.05）。SEA和SWA均促进滋养细胞分泌组织抑制基质金属蛋白酶-1（tissue inhibitor of metalloproteinases-1, TIMP-1），分泌量为（1 925.93±143.22）pg/ml和（1 749.83±53.91）pg/ml（t=4.417，4.402，P均〈0.05）。 结论 本研究采用日本血吸虫不同虫期的抗原作用人早孕原代滋养细胞，发现ES和SEA促进滋养细胞分泌炎症因子；SWA明显促进TGF-β1和TIMP-1分泌。这些结果表明日本血吸虫不同时期的抗原影响人早孕期的滋养细胞功能，为加大疫区育龄妇女妊娠期间的监测提供新的线索。

Objective To explore whether Schistosoma japonicum antigens can affect first trimester human trophoblast cells for providing new clues towards clinical parasite control of women during the process of pregnancy in epidemic areas. Methods Human placental tissues were collected by artificial abortion at 6- 10 weeks of gestation. The trophoblast cells were isolated by proteinase digestion and cultured in vitro. The supernatants were harvested respectively from the cultured cells after having been treated with 25 μg/ml excretory/ secretory antigens from egg（ES）, soluble egg antigens（SEA） and soluble worm antigens（SWA） from S.japonicum for 18 h. The level of cytokines in the culture supernatants was measured by ProcartaPlex detection based on enzyme-linked immunosorbent assay（ELISA）. Results It is found that ES could significantly promote the secretion of IL-6（155.58±20.07） pg/ml（t=8.472, P〈0.05） and IL-8（12.78±2.35） ns/ml（t=8.006, P〈0.05） , while SEA could significantly increase the production of IL-8 （14,18±2.24） ns/ml （t=7.413, P〈 0.05）. SEA could also increase the production of monoeyte ehemotaotic protein 1 （MCP-1）（212.2±14.57） pg/ml （t=15.640, P〈0.05）. The levels of TGF-β1 in the presence of ES（78.56±5.71） ps/ml and SWA（66.61±5.55） pg/ml were significantly hisher than that of medium control. The secretory tissue inhibitor of metalloproteinases-1 （TIMP-1） was increased significantly in the presence of SEA（1 925.93±143.22） pg/ml and SWA （ 1 749.83±53.91 ） pg/ml, compared with the medium group. Conclusion This is the first time for exploring the effects of antigens from different development stages of S. japonicum on cytokines secretion of primary-cultured first trimester human trophoblast cells. We found that ES and SEA promoted the secretion of inflammatory oytokines in trophoblast cells. In addition, SWA significantly promoted TGF-β1 and TIMP-1 secretion. These results suggnst that the antisens of .S. japonicum during the different development stages affect the function of first trimester human trophoblast cells, thus to provide new clues of subsequent pregnancy monitoring in schistosomiasis epidemic areas.