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目的分析细粒棘球蚴感染小鼠单核髓源抑制性细胞(M-MDSC)精氨酸酶的表达和活性变化。方法 12只BALB/c雌性小鼠随机分为对照组和感染组。感染组小鼠腹腔注射活原头节2 000个/只,对照组小鼠注射等体积生理盐水。于感染后120 d采集小鼠眼眶静脉丛外周血,解剖并观察小鼠腹腔和各脏器的病变。无菌取小鼠脾组织,制备单细胞悬液后,利用免疫磁珠分离M-MDSC。提取并纯化M-MDSC RNA,合成c DNA,芯片检测筛选感染组和对照组的M-MDSC差异基因,荧光定量PCR验证差异基因的表达。精氨酸酶检测试剂盒检测小鼠外周血中的精氨酸酶活性。结果BALB/c小鼠感染细粒棘球蚴原头节后120 d,腹腔和内脏器官中形成单性包囊。免疫磁珠分离获得M-MDSC。芯片杂交和荧光定量PCR检测结果显示,感染组小鼠M-MDSC源精氨酸酶相对表达量分别为7.92±0.85和11.97±5.39,均高于对照组小鼠(1.65±0.19和1.00±0.57)(P<0.05)。检测结果表明,感染组小鼠外周血中的精氨酸酶活性为(3.83±0.44)U/L,高于对照组小鼠[(1.57±0.57)U/L](P<0.05)。结论细粒棘球蚴感染小鼠M-MDSC精氨酸酶的表达量和活性显著升高。
Objective To analyze the changes of arginase expression and activity in mononuclear marrow derived suppressor cells (M-MDSC) infected with Echinococcus granulosus. Methods Twelve BALB / c female mice were randomly divided into control group and infected group. Infected mice were injected intraperitoneally with 2 000 live head segments, and control mice were injected with equal volume of normal saline. The orbital venous plexus peripheral blood was collected at 120 days after infection and the lesions in the abdominal cavity and various organs of the mice were dissected and observed. Mice were aseptically spleen tissue, single cell suspension preparation, the use of magnetic beads isolated M-MDSC. M-MDSC RNA was extracted and purified, c DNA was synthesized, and the differentially expressed genes of M-MDSC in infected and control groups were screened by microarray. Fluorescent quantitative PCR was used to verify the differential gene expression. Arginase detection kit for detecting arginase activity in mouse peripheral blood. Results BALB / c mice were infected with Echinococcus granulosus 120 days after the formation of the cytoplasm in the peritoneal cavity and internal organs formed a single cyst. Immunomagnetic separation of M-MDSC. The results of chip hybridization and real-time PCR showed that the relative expression of M-MDSC-derived arginase in infected mice was 7.92 ± 0.85 and 11.97 ± 5.39, respectively, which were significantly higher than those in control mice (1.65 ± 0.19 and 1.00 ± 0.57 ) (P <0.05). The results showed that the arginase activity in the peripheral blood of the infected mice was (3.83 ± 0.44) U / L, higher than that of the control mice [(1.57 ± 0.57) U / L] (P <0.05). Conclusion The expression and activity of M-MDSC arginase in mice infected with Echinococcus granulosus were significantly increased.