α-半乳糖酰基鞘胺醇诱发小鼠流产机制初探

Mechanism of α-galactosylceramide Induced Abortion in Mice

目的利用α-半乳糖酰基鞘胺醇(α-galactosylceramide,α-Galcer)诱发性C57BL/6J小鼠流产模型,初步探讨NKT细胞活化与C57BL/6J小鼠流产的关系。方法孕7 d和孕10 d C57BL/6J小鼠,每组8只腹腔注射α-Galcer,72 h后处死鼠剖腹探察胚胎数量。于0,24 h取胎盘淋巴细胞,流式细胞术检测CD3+NK1.1+细胞百分率和CD69+/CD3+NK1.1+细胞百分率,ELISA方法检测α-Galcer处理不同时间血清孕酮(P)水平,并做子宫胎盘免疫组化切片观察。结果C57BL/6J小鼠可由α-Galcer引起流产。胎盘CD3+NK1.1+淋巴细胞在0,24 h分别为(2.96±0.22)%和(4.17±0.67)%,CD69+/CD3+NK1.1+细胞百分率分别为(6.54±1.13)%和(82.45±5.85)%。ELISA方法测得血清P浓度为(0.15±0.01)ng/ml与对照组(1.28±0.03)ng/ml相比差异有显著性。胎盘子宫免疫组化切片显示α-Galcer处理组孕激素受体(PR)表达明显强于对照组。结论α-Galcer可诱发C57BL/6J小鼠流产,NKT细胞的活化及其分泌的细胞因子可能是导致流产的主要原因。

Objective To develop a model of α - Galcer induced abortion in C57BL/6J mice and to investigate the relationship between natural killer T cell activation and abortion. Methods Pregnant C57BL/6J αmice were injected intraperitoneally with α - Galcer. Mice were sacrificed 72 hours later to exmamine the presence of abortion. Placental lymphocytes were collected at Oh and 24h. The ratio of CD3^＋ NK1.1^＋ and CD69^＋ / CD3^＋ NK1.1^＋ cells were determined with flow cytometr. Serum progesterone was determined by ELISA in mice receiving - Galcer treatment for different duration. The placenta was examined by immunohistochemistry. Results α - Galcer induced abortion in C57BL/6J. At Oh and 24th hour, the ratios of CD3^＋NK1.1^＋ lymphocytes in placenta was （2. 96±0. 22）% and （4. 17±0.67）%, and the ratios of CD69^＋/ CD3^＋NK1.1^＋ cells were （6. 54 ± 1.13）% and （82.45 ± 5. 85）%. Serum progesterone was significantly higher than that in the control. The expression of progesterone receptor in mice received α - Galcer administration was significant higher than that in the control. Conclusion α - Galcer induces miscarriage in C57BL/6J αmice, probably associ- ated with NKT activation and cytokine secretion.