经饮水甲基汞对小鼠脾脏中成熟免疫细胞影响的时间-效应特征

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[目的]观察甲基汞暴露后不同时间点B10.S小鼠脾脏中髓系细胞、淋巴系细胞和骨髓中造血祖细胞生成相应成熟髓系细胞集落的数量及其动态变化,解释成熟免疫细胞动态变化的可能原因,探讨甲基汞引起机体免疫紊乱的可能机制。[方法]以6到8周龄的B10.S雌性小鼠为研究对象,随机分为对照组和实验组,分别饮用去离子水、1.25μmol/L甲基汞水溶液4周,测汞仪测定脾脏和脑中汞的含量,每周观察饮水和饮食消耗量。分别在第1周、2周和4周用动物电子称称量小鼠体重,并且在第1周、2周和4周用流式细胞仪检测小鼠脾脏中巨噬细胞、单核细胞、中性粒细胞、B淋巴细胞、CD4~+T细胞、CD8~+T细胞和自然杀伤(NK)细胞的数量,并用细胞集落形成试验(CFU,colony-formation units)检测骨髓造血祖细胞生成成熟髓系细胞的能力。[结果]2组小鼠饮水量、饮食消耗量、体重变化量之间的差异均无统计学意义(P>0.05)。饮用甲基汞溶液1周后,与对照组相比,小鼠脾脏中单核细胞、中性粒细胞在脾脏细胞中的百分比增高;第2周,单核细胞、巨噬细胞、中性粒细胞的数量百分比降低(P<0.05)。与对照组相比,B细胞数量百分比在第2周和第4周增高(P<0.05),CD4~+T细胞的数量百分比在第4周下降,CD8~+T细胞、NK细胞的数量百分比在第2周下降,差异均具有统计学意义(P<0.05)。CFU实验表明,在饮用甲基汞后第1周,粒细胞-红细胞-巨噬细胞-单核细胞集落(GEMM)数量、粒细胞-巨噬细胞集落(GM)数量、粒细胞集落(G)和巨噬细胞集落(M)数量增多,与对照组相比,差异有统计学意义(P<0.05);与对照组相比,饮用甲基汞2周后,GEMM和GM数量下降(P<0.05),4周后G和M数量增多(P<0.05)。[结论]甲基汞可导致小鼠骨髓中髓系祖细胞分化生成相应成熟细胞的能力先增强后减弱再恢复至正常水平,进而导致单核细胞、中性粒细胞、巨噬细胞的百分比呈现先升高后降低再逐渐恢复的趋势,CD4~+T细胞占脾脏细胞的百分比总体呈现下调的趋势,CD8~+T细胞、NK细胞占脾脏细胞的百分比先下调后上升,B细胞占脾脏细胞的比例总体呈现上调的趋势。甲基汞暴露后不同时间点小鼠脾脏中成熟免疫细胞的动态变化很复杂,总体上可以用CFU的变化来解释髓系细胞变化的原因。成熟免疫细胞的数量百分比随染毒时间的变化而发生改变,提示甲基汞引起的免疫紊乱可能与其导致的成熟免疫细胞比例失调有关。 [Objective] To observe the number and dynamic changes of myeloid progenitor cells in myeloid, lymphoid cells and bone marrow of splenic B10 · S mice exposed to methylmercury at different time points to explain the maturation of myeloid cells. The possible reasons for the dynamic changes and to explore the possible mechanism of methylmercury-induced immune disorders. [Method] Sixty-eight weeks old B10.S female mice were randomly divided into control group and experimental group, which were respectively treated with deionized water and 1.25μmol / L methylmercury aqueous solution for 4 weeks. Measured by mercury analyzer Spleen and brain mercury content, weekly drinking water and diet consumption. The body weight of mice was weighed by animal electron at the first week, the second week and the fourth week, respectively. The macrophages, monocytes, The number of neutrophils, B lymphocytes, CD4 ~ + T cells, CD8 ~ + T cells and natural killer (NK) cells were measured and the maturation of bone marrow hematopoietic progenitor cells was detected by colony forming unit (CFU) Myeloid cells ability. [Result] There was no significant difference between the two groups in drinking water consumption, dietary consumption and weight change (P> 0.05). One week after drinking methylmercury solution, the percentage of monocytes and neutrophils in spleen cells increased in spleen of mice compared with control group. In the second week, monocytes, macrophages, neutrophils The percentage of cells decreased (P <0.05). Compared with the control group, the percentage of B cells increased at week 2 and 4 (P <0.05), while the percentage of CD4 ~ + T cells decreased at week 4. The percentage of CD8 ~ + T cells and NK cells In the second week of decline, the differences were statistically significant (P <0.05). CFU experiments showed that the number of granulocyte-erythrocyte-macrophage-monocyte colony (GEMM), granulocyte-macrophage colony (GM), granulocyte colony (G) (P <0.05). Compared with the control group, the amount of GEMM and GM decreased after drinking methylmercury for two weeks (P <0.05), and the number of macrophage colony (M) increased, compared with the control group 0.05). After 4 weeks, the number of G and M increased (P <0.05). [Conclusion] Methylmercury can increase the ability of myeloid progenitor cells to differentiate into mature cells in the bone marrow of mice, and then increase and then weaken and then return to normal levels, leading to the percentage of monocytes, neutrophils and macrophages The percentage of CD4 ~ + T cells in spleen cells showed an overall downward trend. The percentage of CD8 ~ + T cells and NK cells in spleen cells first decreased and then increased, and the percentage of B cells in spleen cells The proportion of the overall upward trend showed upward trend. The dynamic changes of mature immune cells in mouse spleen at different time points after exposure to methylmercury are complex. The changes of CFU can generally be used to explain the reasons for the changes of myeloid cells. The percentage of mature immune cells changed with the time of exposure, suggesting that the immune disorder caused by methylmercury might be related to the imbalance of mature immune cells.
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