再生复血汤对再生障碍性贫血小鼠外周血CD4^+ CD25^+ CD127^-调节性T细胞及Foxp3 mRNA的影响

Effect of Zaisheng Fuxue Decoction on CD4^+ CD25^+ CD127^- Regulatory T Cells and Transcription Factor Foxp3 mRNA in the Peripheral Blood of Aplastic Anemia Mice

目的探讨再生复血汤对再生障碍性贫血(AA)小鼠外周血CD4^+CD25^+CD127^-调节性T细胞(Treg)及核转录因子Foxp3 mRNA的影响。方法将72只小鼠随机分成空白组,模型组,阳性对照组,再生复血汤高、中、低剂量组,每组12只。除空白组外,其余各组小鼠予^(60)Co-γ联合腹腔注射环磷酰胺和氯霉素建立AA小鼠模型。造模第7天再生复血汤高、中、低剂量组分别给予再生复血汤10、5、2.5 g/(kg·d)灌胃,模型组和空白组予等体积的生理盐水灌胃,阳性对照组予环孢素3 mg/(kg·d)灌胃,各组均干预30天。检测小鼠外周血白细胞数(WBC)、网织红细胞数(Ret)、血红蛋白数(Hb)及血小板数(PLT)、骨髓有核细胞数(粒系比例、红系比例、巨核细胞数,淋巴细胞比例)、Treg比例和单个核细胞Foxp3mRNA表达量。结果与空白组比较,模型组外周血WBC、Ret、Hb、PLT、骨髓粒系比例、红系比例、巨核细胞数、Treg比例、Foxp3 mRNA表达量降低,淋巴细胞比例升高(均P<0.05)。与模型组比较,各干预组小鼠外周血WBC、Ret、Hb、PLT、骨髓粒系比例、红系比例、巨核细胞数、Treg比例、Foxp3 mRNA表达量升高,淋巴细胞比例下降(均P<0.05)。与再生复血汤低剂量组比较,再生复血汤中、高剂量组和阳性对照组外周血WBC、Ret、Hb、PLT、骨髓粒系比例、红系比例、巨核细胞数、Treg比例、Foxp3 mRNA表达量升高,淋巴细胞比例下降(均P<0.0 5),且再生复血汤高剂量组优于再生复血汤中剂量组和阳性对照组(均P<0.05)。结论再生复血汤能通过促使AA小鼠外周血单个核细胞Foxp3 mRNA表达,使得Treg比例升高,诱导AA小鼠恢复自身免疫耐受,并且呈现剂量依赖性。

Objective To explore the effect of Zaisheng Fuxue Decoction on CD4 ＋ CD25 ＋ CD127-regulatory T cell （Treg） and Foxp3 mRNA in the peripheral blood of aplastic anemia（AA） mice. Methods Seventy-two mice were randomly divided into the blank group, the model group, the positive control group and Zaisheng Fuxue Decoction （low dose, middle dose, and high dose） groups, 12 in each group. In addition to the blank group, mice of the other groups were treated with ^60Co-γcombined with in- traperitoneal injection of cyclophosphamide and chloramphenicol to establish AA mouse model. At the seventh day of modeling, the high does, middle dose and low dose of Zaisheng Fuxue Decoction groups of AA mice were intragastric administrated with the decoction at the dose of 10, 5 and 2.5 g/（kg · d）, separately; the model group and the blank group were given an equal volume of saline, and the positive control group was treated with cyclosporine of 3 mg/（kg· d）. All groups were treated for 30 days. The white blood cell （WB C） count, reticolociti （ Ret）, hem oglobin counts （ Hb）, platelet count （ PLT）, nucleat- ed cells in bone marrow（the portion of granule count, the portion of red cells, megakaryocytes, the por- tion of lymphocytes） were measured . The proportion of Treg and the mRNA expression of Foxp3 in the peripheral blood mononuclear cells of each group were detected. Results Compared with the blank group, the WBC count, Ret, Hb and PLT level, the portion of bone marrow granule, red cells and megakaryo- cytes, the proportion of Treg cells, the mRNA expression of Foxp3 of the peripheral blood in the model group decreased significantly, and the portion of lymphocytes increased significantly （P 〈 0.05）. Com- pared with low dose Zaisheng Fuxue Decoction group, the WBC count, Ret, Hb and PLT level increased, the portion of bone marrow granule, red cells and megakaryocytes, the portion of Treg and the mRNA expression of Foxp3 increased, the portion of lymphocytes decreased（P 〈0.05）, and the therapeutic re- sponse of the high dose Zaisheng Fuxue Decoction group was better than that of the middle dose group and the positive control group（P 〈0.05）. Conclusion Zaisheng Fuxue Decoction can promote the mRNA expression of Foxp3 of peripheral blood mononuclear cells in AA mice, increase the proportion of Treg, induce AA mice to restore self-immunization tolerance with the tendency of dose-dependent pattern.