再生胶囊对再生障碍性贫血大鼠外周血CD4~+、CD25~+调节性T细胞及细胞因子IL-10、TGF-β_1的影响
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摘要
目的:探讨再生胶囊对再生障碍性贫血大鼠外周血CD4+、CD25+调节性T细胞(Treg)、核转录因子Foxp3表达及细胞因子IL-10、TGF-β1的影响。方法:将80只SD大鼠随机分成空白组、模型组、阳性对照组和再生胶囊组,每组20只。除空白组外,其余各组大鼠予60Co-γ联合腹腔注射环磷酰胺和氯霉素建立再生障碍性贫血大鼠模型。于造模第7天再生胶囊组给予再生胶囊[0.216 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 d。检测大鼠外周血白细胞数(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、淋巴细胞比例、巨核细胞数),细胞因子IL-10和TGF-β1浓度,Treg细胞比例和单个核细胞Foxp3 mRNA表达量。结果:与空白组比较,模型组大鼠外周血WBC、Ret、Hb、PLT、骨髓粒系比例、红系比例、巨核细胞数、IL-10浓度、TGF-β1浓度、Treg细胞比例、Foxp3 mRNA表达量降低,淋巴细胞比例升高(P<0.05)。与模型组比较,再生胶囊组和阳性对照组大鼠外周血WBC、Ret、Hb、PLT、骨髓粒系比例、红系比例、巨核细胞数、IL-10浓度、TGF-β1浓度、Treg细胞比例、Foxp3 mRNA表达量升高,淋巴细胞比例下降(P<0.05)。与阳性对照组比较,再生胶囊组大鼠外周血WBC、Ret、Hb、PLT、骨髓粒系比例、红系比例、巨核细胞数、IL-10浓度、TGF-β1浓度、Treg细胞比例、Foxp3mRNA表达量升高,淋巴细胞比例下降(P<0.05)。结论:再生胶囊能通过促使再生障碍性贫血大鼠外周血Treg细胞数和FOXP3表达量增多,产生抑制性细胞因子IL-10和TGF-β1,恢复自身免疫耐受平衡,改善骨髓造血衰竭。
Objective:To explore the effect of Zaisheng capsule on CD4+CD25+-regulatory T cell(Treg),transcription factor Foxp3 mRNA and cytokines IL10/TGF-β1 in the peripheral blood of aplastic anemia(AA)rats.Methods:80 rats were randomly divided into the blank control group,the model group,the positive control group and Zaisheng capsule group,20 in each group.In addition to the blank control group,the other groups of rats were treated with60 Co-γcombined via intraperitoneal injection of cyclophosphamide and chloramphenicol to establish AA rat model.At the seventh day of modeling,Zaisheng capsule group of AA rat were intragastric administrated with the decoction at the dose of 0.216 g/(kg·d),separately;the model group and the normal rat 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 count(WBC),reticolociti(Ret),hemoglobin counts(Hb),platelet count(PLT),nucleated cells in bone marrow(the portion of granule count,the portion of red cells,the portion of lymphocytes,megakaryocytes) were measured.Cytokines IL-10,TGF-β1,proportion of Treg cell and the expression of mRNA Foxp3 in the peripheral blood mononuclear cells of each group were detected.Results:Compared with the blank controller group,the WBC counting,Ret,Hb and PLT level,the portion of bone marrow granule,red cells and megakaryocytes,cytokines IL-10,TGF-β1,the proportion of Treg cells,the expression of Foxp3 mRNA of the peripheral blood in the model group decreased significantly,and the portion of lymphocytes increased significantly(P<0.05).Compared with the model group,the WBC counting,Ret,Hb and PLT level,the portion of bone marrow granule,red cells and megakaryocytes,cytokines IL-10,TGF-β1,the proportion of Treg cells,the expression of Foxp3 mRNA of the peripheral blood in the Zaisheng capsule group and positive group increased significantly,and the portion of lymphocytes decreased significantly(P <0.05).Compared with the positive group,the WBC counting,Ret,Hb and PLT level,the portion of bone marrow granule,red cells and megakaryocytes,cytokines IL-10,TGF-β1,the proportion of Treg cells,the expression of Foxp3 mRNA of the peripheral blood in the Zaisheng capsule group increased significantly,and the portion of lymphocytes decreased significantly(P<0.05).Conclusions:Zaisheng capsule can promote the proportion of Treg cell and expression of Foxp3 mRNA of peripheral blood in AA rat,produce inhibitory cytokines IL-10 and TGF-β1,restore the balance of autoimmune tolerance,and improve the hematopoietic failure of bone marrow.
Objective:To explore the effect of Zaisheng capsule on CD4+CD25+-regulatory T cell(Treg),transcription factor Foxp3 mRNA and cytokines IL10/TGF-β1 in the peripheral blood of aplastic anemia(AA)rats.Methods:80 rats were randomly divided into the blank control group,the model group,the positive control group and Zaisheng capsule group,20 in each group.In addition to the blank control group,the other groups of rats were treated with60 Co-γcombined via intraperitoneal injection of cyclophosphamide and chloramphenicol to establish AA rat model.At the seventh day of modeling,Zaisheng capsule group of AA rat were intragastric administrated with the decoction at the dose of 0.216 g/(kg·d),separately;the model group and the normal rat 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 count(WBC),reticolociti(Ret),hemoglobin counts(Hb),platelet count(PLT),nucleated cells in bone marrow(the portion of granule count,the portion of red cells,the portion of lymphocytes,megakaryocytes) were measured.Cytokines IL-10,TGF-β1,proportion of Treg cell and the expression of mRNA Foxp3 in the peripheral blood mononuclear cells of each group were detected.Results:Compared with the blank controller group,the WBC counting,Ret,Hb and PLT level,the portion of bone marrow granule,red cells and megakaryocytes,cytokines IL-10,TGF-β1,the proportion of Treg cells,the expression of Foxp3 mRNA of the peripheral blood in the model group decreased significantly,and the portion of lymphocytes increased significantly(P<0.05).Compared with the model group,the WBC counting,Ret,Hb and PLT level,the portion of bone marrow granule,red cells and megakaryocytes,cytokines IL-10,TGF-β1,the proportion of Treg cells,the expression of Foxp3 mRNA of the peripheral blood in the Zaisheng capsule group and positive group increased significantly,and the portion of lymphocytes decreased significantly(P <0.05).Compared with the positive group,the WBC counting,Ret,Hb and PLT level,the portion of bone marrow granule,red cells and megakaryocytes,cytokines IL-10,TGF-β1,the proportion of Treg cells,the expression of Foxp3 mRNA of the peripheral blood in the Zaisheng capsule group increased significantly,and the portion of lymphocytes decreased significantly(P<0.05).Conclusions:Zaisheng capsule can promote the proportion of Treg cell and expression of Foxp3 mRNA of peripheral blood in AA rat,produce inhibitory cytokines IL-10 and TGF-β1,restore the balance of autoimmune tolerance,and improve the hematopoietic failure of bone marrow.
引文
[1]邓德君,曾清,赖春凤,等.再生障碍性贫血细胞免疫机制的研究进展[J].广西医学,2017,39(8):1215-1218.
[2]Kordasti S,Costantini B,Seidl T,et al.Deep phenotyping of Tregs identifies an immune signature for idiopathic aplastic anemia and predicts response to treatment[J].Blood,2016,128(9):1193-1205.
[3]Dao A T,Yamazaki H,Takamatsu H,et al.Cyclosporine restores hematopoietic function by compensating for decreased Tregs in patients with pure red cell aplasia and acquired aplastic anemia[J].Ann Hematol,2016,95(5):771-781.
[4]江劲波,袁通春,赵复锦,等.再生复血汤治疗慢性再生障碍性贫血80例临床观察[J].湖南中医药大学学报,2008,28(3):51-53.
[5]刘璇,王跃,江劲波.江劲波应用再生复血汤治疗再生障碍性贫血经验[J].山西中医,2012,28(2):4-5.
[6]章海斌,王小中,李静.再生障碍性贫血大鼠模型的制作及实验室评价[J].中国实验诊断学,2009,13(10):1360-1362.
[7]Zhang H F,Huang Z D,Wu X R,et al.Comparison of T lymphocyte subsets in aplastic anemia and hypoplastic myelodysplastic syndromes[J].Life Sci,2017,189(15):71-75.
[8]Yan L,F u R,Liu H,et al.Abnormal quantity and function of regulatory T cells in peripheral blood of patients with severe aplastic anemia[J].Cell Immunol,2015,296(2):95-105.
[9]刘永华,江锦红,吴迪炯,等.补肾活血法与益气补肾法治疗慢性再生障碍性贫血的疗效比较[J].中国中西医结合急救杂志,2015,22(3):234-237.
[10]Geissler K,Kabrna E,Kollars M,et al.Interleukin-10 inhibits in vitro hematopoietic suppression and production of interferon-gamma and tumor necrosis factoralpha by peripheral blood mononuclear cells from patients with aplastic anemia[J].Hematol J,2012,3(4):206-213.
[11]齐薇薇,付蓉,王化泉,等.重型再生障碍性贫血患者外周血调节性T细胞的数量异常及其临床意义[J].中国实验血液学杂志,2014,22(4):1043-1046.
[12]In J W,Lee N,Roh E Y,et al.Association of aplastic anemia and FoxP3 gene polymorphisms in Koreans[J].Hematology,2017,22(3):149-154.
[13]王旭丽,林赠华,刘红,等.鹿龙再生汤对重型再生障碍性贫血小鼠CD4+CD25+调节性T细胞内FOXP3表达的影响[J].中华血液学杂志,2015,36(8):689-691.
[14]曹大春.中药黄芪药理作用及临床应用分析[J].医药前沿,2017,7(20):34-35.
[15]毛坤,夏新中,张虎.中药石韦的药理作用与临床应用研究进展[J].长江大学学报,2014,11(6):110-112.
[2]Kordasti S,Costantini B,Seidl T,et al.Deep phenotyping of Tregs identifies an immune signature for idiopathic aplastic anemia and predicts response to treatment[J].Blood,2016,128(9):1193-1205.
[3]Dao A T,Yamazaki H,Takamatsu H,et al.Cyclosporine restores hematopoietic function by compensating for decreased Tregs in patients with pure red cell aplasia and acquired aplastic anemia[J].Ann Hematol,2016,95(5):771-781.
[4]江劲波,袁通春,赵复锦,等.再生复血汤治疗慢性再生障碍性贫血80例临床观察[J].湖南中医药大学学报,2008,28(3):51-53.
[5]刘璇,王跃,江劲波.江劲波应用再生复血汤治疗再生障碍性贫血经验[J].山西中医,2012,28(2):4-5.
[6]章海斌,王小中,李静.再生障碍性贫血大鼠模型的制作及实验室评价[J].中国实验诊断学,2009,13(10):1360-1362.
[7]Zhang H F,Huang Z D,Wu X R,et al.Comparison of T lymphocyte subsets in aplastic anemia and hypoplastic myelodysplastic syndromes[J].Life Sci,2017,189(15):71-75.
[8]Yan L,F u R,Liu H,et al.Abnormal quantity and function of regulatory T cells in peripheral blood of patients with severe aplastic anemia[J].Cell Immunol,2015,296(2):95-105.
[9]刘永华,江锦红,吴迪炯,等.补肾活血法与益气补肾法治疗慢性再生障碍性贫血的疗效比较[J].中国中西医结合急救杂志,2015,22(3):234-237.
[10]Geissler K,Kabrna E,Kollars M,et al.Interleukin-10 inhibits in vitro hematopoietic suppression and production of interferon-gamma and tumor necrosis factoralpha by peripheral blood mononuclear cells from patients with aplastic anemia[J].Hematol J,2012,3(4):206-213.
[11]齐薇薇,付蓉,王化泉,等.重型再生障碍性贫血患者外周血调节性T细胞的数量异常及其临床意义[J].中国实验血液学杂志,2014,22(4):1043-1046.
[12]In J W,Lee N,Roh E Y,et al.Association of aplastic anemia and FoxP3 gene polymorphisms in Koreans[J].Hematology,2017,22(3):149-154.
[13]王旭丽,林赠华,刘红,等.鹿龙再生汤对重型再生障碍性贫血小鼠CD4+CD25+调节性T细胞内FOXP3表达的影响[J].中华血液学杂志,2015,36(8):689-691.
[14]曹大春.中药黄芪药理作用及临床应用分析[J].医药前沿,2017,7(20):34-35.
[15]毛坤,夏新中,张虎.中药石韦的药理作用与临床应用研究进展[J].长江大学学报,2014,11(6):110-112.