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目的研究小鼠合体滋养细胞微泡(STBM)对骨髓源性树突状细胞(BMDCs)免疫功能的影响。方法 2015年于第三军医大学大坪医院野战外科研究所妇产科中心,机械分离法将小鼠胎盘剪碎,采用差速离心法制备合体滋养细胞微泡。添加重组粒细胞巨噬细胞集落刺激因子(rm GM-csf)、重组白细胞介素-4(IL-4)培养骨髓源性树突状细胞,6 d后将其分为阴性对照组和实验组1、2、3、4以及阳性对照组。实验组1、2、3、4分别加入终浓度为1.0、1.5、2.0、2.5 mg/L的STBM,阴性对照组加等量的磷酸盐缓冲液(PBS),阳性对照组加入2.0 mg/L的脂多糖(LPS),继续培养2 d。流式细胞学技术(FCM)检测树突状细胞表面标志物CD11c及CD86的表达情况。同时ELISA法检测不同浓度STBM诱导下树突状细胞(dendritic cells,DCs)分泌肿瘤坏死因子-α(TNF-α),干扰素-γ(IFN-γ)及IL-12的含量。结果不同浓度STBM处理后的BMDCs明显上调了CD11c、CD86的表达,其中1.0、1.5、2.0、2.5 mg/L的STBM诱导DCs的CD11c/CD86双阳性表达率分别为46.1%、56.9%、60.5%、88.9%,与PBS组(18.2%)相比,差异具有统计学意义(P<0.05)。不同浓度的STBM刺激DCs后产生的TNF-α[(6.91±1.37)、(7.98±1.55)、(9.40±0.93)、(12.65±1.62)ng/L],IFN-γ[(12.14±1.39)、(17.52±1.67)ng/L]及IL-12[(6.98±1.19)、(7.88±0.10)、(9.18±1.69)ng/L]分别与各PBS组[(3.61±0.64)、(3.15±0.90)、(3.51±1.09)ng/L]相比,差异有统计学意义(P<0.05)。结论 STBM促进DCs的成熟,激活母体系统性炎症反应,可能是引起子痫前期发生重要原因。
Objective To investigate the effect of mouse syncytiotrophoblastic microbubbles (STBM) on the immune function of bone marrow-derived dendritic cells (BMDCs). Methods In 2015, the placenta of mice were cut off by mechanical separation method and the syncytiotrophoblastic microbubbles were prepared by differential centrifugation in the Obstetrics and Gynecology Center of Daping Hospital, Third Military Medical University. After adding recombinant human granulocyte-macrophage colony-stimulating factor (rm GM-csf) and recombinant interleukin-4 (IL-4) to culture bone marrow-derived dendritic cells, they were divided into negative control group and experimental group 1,2,3,4 and positive control group. The experimental groups 1, 2, 3, and 4 were added STBM with final concentrations of 1.0, 1.5, 2.0 and 2.5 mg / L respectively. The negative control group was given phosphate buffered saline (PBS) Lipopolysaccharide (LPS), continue to cultivate 2 d. Flow cytometry (FCM) was used to detect the expression of dendritic cell surface markers CD11c and CD86. Meanwhile, the levels of tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IL-12 secreted by dendritic cells (DCs) induced by different concentrations of STBM were detected by ELISA. Results The expressions of CD11c and CD86 were significantly up-regulated by STBM-treated BMDCs. The positive rates of CD11c / CD86 double staining in STBM-induced DCs were 46.5%, 56.9% and 60.5% , 88.9% compared with PBS group (18.2%), the difference was statistically significant (P <0.05). The levels of TNF-α [(6.91 ± 1.37), (9.80 ± 1.55), (12.65 ± 1.62) ng / L] and IFN-γ [(12.14 ± 1.39) , (3.61 ± 0.64) and (3.15 ± 0.64) ng / L, respectively, compared with PBS group (17.52 ± 1.67 ng / L) and IL-12 [6.98 ± 1.19 and 7.88 ± 0.10 and 9.18 ± 1.69 ng / ± 0.90), (3.51 ± 1.09) ng / L], the difference was statistically significant (P <0.05). Conclusion STBM can promote the maturation of DCs and activate maternal systemic inflammatory response, which may be one of the important causes of preeclampsia.