益髓生血颗粒对AA大鼠CD4 + CD25 + Foxp3调节性T细胞的影响

目的：观察益髓生血颗粒对再生障碍性贫血（Aplastic Anemia,AA）大鼠CD4 + CD25 + 调节性T细胞（Regulatory T Cell,Treg细胞）的影响及其治疗AA的作用机制。方法：取雄性SD大鼠按体重随机分组。其中模型组连续7周隔天皮下注射苯（1 mL ? kg -1 ）、取材10天前开始腹腔注射环磷酰胺（25 mL ? kg -1 ）,连续3天,第4周将模型组大鼠按体重随机分为模型组、司坦唑醇组、益髓生血颗粒组,灌胃给药。正常组及模型对照组给予等体积生理盐水。实验结束时,检测外周血WBC、RBC、HGB、PLT;制备血涂片、骨髓涂片;免疫组织化学法检测脾组织Treg细胞Foxp3蛋白表达;RT-PCR法检测骨髓组织Foxp3 mRNA的表达。结果：与正常组比较,模型组WBC、RBC、HGB、PLT均显著减少（P<0.01）,血涂片示血细胞通透性差、白细胞减少、退化细胞增多,骨髓涂片示脂肪滴明显增加、造血细胞均明显降低、非造血细胞增多;经益髓生血颗粒组治疗后WBC、RBC、HGB、PLT均显著增加（P<0.01）,血涂片及骨髓涂片显示细胞通透性好转、形态趋于正常、脂肪滴明显减少,造血细胞均明显增加,脾组织Foxp3蛋白及骨髓组织Foxp3 mRNA表达明显升高（P<0.01）。结论：益髓生血颗粒可上调Foxp3的因及蛋白表达,调控AA免疫功能,进而改善AA免疫环境,促进骨髓造血功能的恢复,发挥治疗AA的重要作用。

Effect of Yisui Shengxue Granules on CD4 + CD25 + Foxp3 Regulatory T Cells ofAplasticAnemia in Rats

This paper was aimed to observe the effect of Yisui Shengxue (YSSX) granules on CD4 + CD25 + regulatory T cells (Treg cells) and its treatment mechanism in aplastic anemia (AA) rats. Male SD rats were selected and randomly divided into different groups according to their weight. In the model group, subcutaneous injection of benzene (1 mL ? kg -1 )was given every other day for 7 consecutive weeks. Ten days before the rats were sacrificed, intraperitoneal injection of CTX (25 mL ? kg - 1 ) was given for 3 consecutive days. On the 4 th week, model rats were divided into the model group, stanozolol group, and the YSSX granules group. Intragastric administration of corresponding drug was given. Same volume of normal saline was given to the normal group and the model group. At the end of the experiment, WBC, RBC,HGB and PLT in peripheral blood were detected. Blood smear and bone marrow smear were prepared. The Foxp3 protein expression of Treg cells in spleen tissues was detected by immunohistochemistry (IHC). RT-PCR was used to detect the Foxp3 mRNA expression in bone marrow tissues. The results showed that compared with the normal group, WBC, RBC,HGB and PLT in the model group were significantly reduced (P < 0.01). The blood smear showed poor permeability of blood cells, reduced WBCs, and increased degenerated cells. The bone marrow smear indicated significantly increased fat drops, significantly reduced hematopoietic cells, and increased nonhematopoietic cells. After the treatment of YSSX granules, WBC, RBC, HGB and PLT were significantly increased (P < 0.01). Both the blood smear and bone marrow smear showed cell permeability improvement, cell form returns to normal, fat drops significantly reduced, significantly increased hematopoietic cells, significantly increased Foxp3 protein expression in spleen tissues and Foxp3 mRNA expression in bone marrow tissues (P < 0.01). It was concluded that YSSX granules can upregulate both gene and protein expression of Foxp3, regulate AA immune function in order to improve the AA immune environment, promote the recovery of bone marrow hematopoietic function, which played an important role in AA treatment.