犀角地黄汤对免疫性血小板减少症模型大鼠外周血CD4~+ CD25~+调节性T细胞及Th17细胞的影响
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摘要
目的探讨犀角地黄汤对免疫性血小板减少症(ITP)模型大鼠外周血CD4~+ CD25~+调节性T细胞(Treg)、Th17细胞及相关细胞因子的影响。方法将48只大鼠随机分为正常组、模型组、醋酸泼尼松组及犀角地黄汤低、中、高剂量组,每组8只。除正常组外,其余组均采用腹腔注射兔抗大鼠血小板血清(APS)建立ITP模型,于造模开始后第7天开始,各给药组灌胃给予相应剂量药物,连续14 d。采用血细胞计数仪检测造模前、灌胃前、灌胃结束后各组大鼠外周血中血小板数目;灌胃结束后,采用流式细胞术检测各组大鼠Treg、Th17细胞比例,采用实时荧光定量PCR法检测淋巴细胞Foxp3 mRNA相对表达量,采用酶联免疫吸附法检测血清白细胞介素-17(IL-17)、IL-35水平。结果灌胃结束后,与模型组比较,犀角地黄汤各剂量组大鼠血小板数目明显上升(P均<0.05),外周血淋巴细胞中Treg所占比例、Foxp3 mRNA相对表达量及IL-35水平均明显上升(P均<0.05),而Th17细胞所占比例及IL-17水平均明显下降(P均<0.05),且具有剂量依赖性;犀角地黄汤高剂量组各指标与醋酸泼尼松组比较差异均无统计学意义(P均>0.05)。结论犀角地黄汤可能通过上调ITP大鼠外周血中Treg细胞表达,下调Th17细胞表达,维持Treg/Th17平衡,进而调节相关细胞因子IL-17、IL-35的表达发挥抗血小板减少作用。
Objective It is to investigate the effect of Xijiao Dihuang Decoction( XJDHD) on CD4~+ CD25~+ regulatory T cells( Treg),Th17 cells and the related cytokines in peripheral blood of rats with immune thrombocytopenia( ITP).Methods Forty-eight rats were randomly divided into normal group,model group,prednisolone acetate group and low dose,middle dose and high dose of XJDHD groups,there were 8 cases in each group. The ITP model was established by treated with intraperitoneal injection of rabbit platelet serum( APS). On the seventh days after the model established,the animals were given the corresponding drugs by gavage of stomach respectively,continuous for 14 days. The number of platelets in peripheral blood was detected by flow cytometry,the percentage of Treg and Th17 cells before and after administration were measured by flow cytometry,the relative expression of Foxp3 mRNA was detected by real-time fluorescence quantitative PCR,and the levels of IL-17 and IL-35 in plasma were detected by enzyme-linked immunosorbent assay( ELISA). Results Compared with the model group,the number of platelets in all dose of XJDHD groups were increased significantly( all P <0. 05),and the proportion of Treg in the peripheral blood lymphocytes,the relative expression of Foxp3 mRNA and the level of IL-35 were significantly increased( all P < 0. 05),and the proportion of Th17 cells and the level of IL-17 were significantly decreased( all P < 0. 05),which showed dose-dependent. There was no significant difference in all the indexes between the high dose of XJDHD group and the prednisone acetate group( P > 0. 05),but there was still significant difference with normal control group( P < 0. 05). Conclusion XJDHD may play the role of antiplatelet reduction,by up regulate the expression of Terg cells in the peripheral blood of ITP rats,down regulation the expression of Th17 cell,maintain the balance between Terg and Th17,and regulate the expression of related cytokines IL-17 and IL-35.
Objective It is to investigate the effect of Xijiao Dihuang Decoction( XJDHD) on CD4~+ CD25~+ regulatory T cells( Treg),Th17 cells and the related cytokines in peripheral blood of rats with immune thrombocytopenia( ITP).Methods Forty-eight rats were randomly divided into normal group,model group,prednisolone acetate group and low dose,middle dose and high dose of XJDHD groups,there were 8 cases in each group. The ITP model was established by treated with intraperitoneal injection of rabbit platelet serum( APS). On the seventh days after the model established,the animals were given the corresponding drugs by gavage of stomach respectively,continuous for 14 days. The number of platelets in peripheral blood was detected by flow cytometry,the percentage of Treg and Th17 cells before and after administration were measured by flow cytometry,the relative expression of Foxp3 mRNA was detected by real-time fluorescence quantitative PCR,and the levels of IL-17 and IL-35 in plasma were detected by enzyme-linked immunosorbent assay( ELISA). Results Compared with the model group,the number of platelets in all dose of XJDHD groups were increased significantly( all P <0. 05),and the proportion of Treg in the peripheral blood lymphocytes,the relative expression of Foxp3 mRNA and the level of IL-35 were significantly increased( all P < 0. 05),and the proportion of Th17 cells and the level of IL-17 were significantly decreased( all P < 0. 05),which showed dose-dependent. There was no significant difference in all the indexes between the high dose of XJDHD group and the prednisone acetate group( P > 0. 05),but there was still significant difference with normal control group( P < 0. 05). Conclusion XJDHD may play the role of antiplatelet reduction,by up regulate the expression of Terg cells in the peripheral blood of ITP rats,down regulation the expression of Th17 cell,maintain the balance between Terg and Th17,and regulate the expression of related cytokines IL-17 and IL-35.
引文
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[3]栾大丽,马桐生.犀角地黄汤治疗小儿过敏性紫癜临床研究[J].河南中医,2016,36(4):653-655
[4]夏乐敏,崔乐乐,姜一陵,等.犀角地黄汤通过线粒体途径抑制免疫性骨髓衰竭血小板凋亡的实验研究[J].江苏中医药,2017,49(4):74-77
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[6]翟菊萍,何杨,杨炳华.免疫失耐受在ITP自身抗体产生机制中的研究进展[J].中华微生物学和免疫学杂志,2016,36(10):790-794
[7]孟亚红,孙丽华.CD4+CD25+调节性T细胞在ITP患者中的表达及与临床预后的相关性[J].浙江临床医学,2017,19(3):446-448
[8]赵海丰,张翼鷟.CD4+CD25+调节性T细胞在免疫性血小板减少症发病中的研究进展[J].天津医药,2014,42(4):390-392
[9]Yu S,Liu C,Li L,et al.Inactivation of Notch signaling reverses the Th17/Treg imbalance in cells from patients with immune thrombocytopenia[J].Lab Invest,2015,95(2):157-167
[10]Talaat RM,Mohamed SF,Bassyouni IH,et al.Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus(SLE)patients:Correlation with disease activity[J].Cytokine,2015,72(2):146-153
[11]贾瑞环,牛红青,李小峰.类风湿关节炎患者外周血及关节液中Treg细胞和Th17细胞水平的研究进展[J/CD].中华临床医师杂志:电子版,2016,10(24):3823-3826
[12]高勇,宋月,韩姝坤,等.Th19、Th17、CD4+CD25+Foxp3+调节性T细胞亚群在成人原发免疫性血小板减少症发病中的意义[J].中华微生物学和免疫学杂志,2016,36(7):500-505
[13]Curtis BR.Recent progress in understanding the pathogenesis of fetal and neonatal alloimmune thrombocytopenia[J].Br J Haematol,2016,171(5):671-682
[14]沈静涵,周韶虹.原发免疫性血小板减少症的中医药治疗进展[J].天津药学,2015,27(6):68-69
[15]张晓蒙,李琳琳.犀角地黄汤联合匹多莫德治疗儿童过敏性紫癜效果及对免疫功能的影响[J].现代中西医结合杂志,2016,25(35):3906-3908
[16]刘爱民,崔利莎,张步鑫,等.麻防犀角地黄汤与犀角地黄汤治疗冬季血热型银屑病的疗效观察[J].中国皮肤性病学杂志,2016,30(6):629-630
[17]王洪彬,崔建美,赵舒,等.基于数据挖掘的中药防治系统性红斑狼疮组方规律研究[J].中国中医基础医学杂志,2016,22(7):970-972
[2]王伟涛,吴深涛,刘宝山.免疫性血小板减少症的中医病因病机浅析[J].中医药学报,2016,44(2):8-9
[3]栾大丽,马桐生.犀角地黄汤治疗小儿过敏性紫癜临床研究[J].河南中医,2016,36(4):653-655
[4]夏乐敏,崔乐乐,姜一陵,等.犀角地黄汤通过线粒体途径抑制免疫性骨髓衰竭血小板凋亡的实验研究[J].江苏中医药,2017,49(4):74-77
[5]吴冰,韦艳,张茜.免疫性血小板减少症治疗进展[J].中国实验血液学杂志,2015,23(4):1212-1215
[6]翟菊萍,何杨,杨炳华.免疫失耐受在ITP自身抗体产生机制中的研究进展[J].中华微生物学和免疫学杂志,2016,36(10):790-794
[7]孟亚红,孙丽华.CD4+CD25+调节性T细胞在ITP患者中的表达及与临床预后的相关性[J].浙江临床医学,2017,19(3):446-448
[8]赵海丰,张翼鷟.CD4+CD25+调节性T细胞在免疫性血小板减少症发病中的研究进展[J].天津医药,2014,42(4):390-392
[9]Yu S,Liu C,Li L,et al.Inactivation of Notch signaling reverses the Th17/Treg imbalance in cells from patients with immune thrombocytopenia[J].Lab Invest,2015,95(2):157-167
[10]Talaat RM,Mohamed SF,Bassyouni IH,et al.Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus(SLE)patients:Correlation with disease activity[J].Cytokine,2015,72(2):146-153
[11]贾瑞环,牛红青,李小峰.类风湿关节炎患者外周血及关节液中Treg细胞和Th17细胞水平的研究进展[J/CD].中华临床医师杂志:电子版,2016,10(24):3823-3826
[12]高勇,宋月,韩姝坤,等.Th19、Th17、CD4+CD25+Foxp3+调节性T细胞亚群在成人原发免疫性血小板减少症发病中的意义[J].中华微生物学和免疫学杂志,2016,36(7):500-505
[13]Curtis BR.Recent progress in understanding the pathogenesis of fetal and neonatal alloimmune thrombocytopenia[J].Br J Haematol,2016,171(5):671-682
[14]沈静涵,周韶虹.原发免疫性血小板减少症的中医药治疗进展[J].天津药学,2015,27(6):68-69
[15]张晓蒙,李琳琳.犀角地黄汤联合匹多莫德治疗儿童过敏性紫癜效果及对免疫功能的影响[J].现代中西医结合杂志,2016,25(35):3906-3908
[16]刘爱民,崔利莎,张步鑫,等.麻防犀角地黄汤与犀角地黄汤治疗冬季血热型银屑病的疗效观察[J].中国皮肤性病学杂志,2016,30(6):629-630
[17]王洪彬,崔建美,赵舒,等.基于数据挖掘的中药防治系统性红斑狼疮组方规律研究[J].中国中医基础医学杂志,2016,22(7):970-972