白芍总苷对EAT小鼠Treg/Th17免疫平衡的影响
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摘要
目的建立实验性自身免疫性甲状腺炎(EAT)小鼠模型,观察不同剂量白芍总苷(TGP)治疗组小鼠脾细胞中调节性T细胞(Treg)和Th17细胞比例及其功能的变化。方法将EAT模型小鼠随机共分为模型(model)组、高剂量TGP (THD)组、中剂量TGP (TMD)组、低剂量TGP(TLD)组和雷公藤多苷(TII)组共5组;real-time PCR检测脾细胞中Foxp3 mRNA和RORγt mRNA的表达;流式细胞计量术检测脾细胞中Treg和Th17细胞的比例;ELISA检测血清中TGF-β1和IL-17的水平;化学发光法检测血清中抗甲状腺球蛋白抗体(TGAb)的水平。结果 TGP处理组(包括低、中和高剂量组)Th17细胞比例较模型组明显降低(P<0.001);中和高剂量TGP组Foxp3 mRNA表达量较模型组明显增高(P<0.05); Treg细胞比例较模型组明显增高(P<0.01,P<0.05);IL-17浓度较模型组明显降低(P<0.01,P<0.05);TGAb浓度较模型组明显降低(P<0.05);中剂量TGP组TGF-β1浓度较模型组显著增高(P<0.01)。结论 TGP能调节Treg/Th17免疫平衡,有利于EAT疾病的转归,其中以中和高剂量TGP组疗效显著。
Objective An experimental autoimmune thyroiditis(EAT) model in mice was established to evaluate the proportion and function of regulatory T cells(Tregs) and Th17 cells in the splenocytes from the treatment group with different doses of total glucosides of paeony(TGP). Methods The EAT model mice were randomly divided into 5 groups: model group, TGP high-dose group, TGP medium-dose group,TGP low-dose group and tripterygium glycosides group. Real-time PCR was performed to detect the expression of Foxp3 mRNA and RORγt mRNA in the splenocytes of each group. Flow cytometry was performed to detect the percentage of Treg and Th17 cells in the mice splenocytes of each group. Serum levels of TGF-β1 and IL-17 in each group were measured by ELISA. Serum level of TGAb in each group was detected by chemiluminescence assay.Results Foxp3 mRNA expression in the mice splenocytes in the medium-dose and in the high-dose TGP group was significantly higher than that in the model group(P< 0.05). The percentages of Treg cells in the medium-dose and the high-dose TGP group were significantly higher than that in the model group( P<0.01,P<0.05). The percentages of T cells secreting IL-17 in the low-, medium-and high-dose groups of TGP were significantly lower than that in the model group(P<0.001). Serum levels of TGF-β1 in the TGP medium-dose group and the TGP high-dose group were significantly higher thanthat in the model group( P<0.01).Serum levels of IL-17 in the TGP medium-dose group and the TGP high-dose group were significantly lower than that in the model group(P<0.01,P<0.05). Serum levels of TGAb in the TGP medium-dose group and the TGP high-dose group were significantly lower than that in the model group(P<0.05). Conclusions TGP can regulate the Treg/Th17 immune balance, which is potentially beneficial to the prognosis of EAT disease. The TGP medium-dose group and the TGP high-dose group have obvious curative effect.
Objective An experimental autoimmune thyroiditis(EAT) model in mice was established to evaluate the proportion and function of regulatory T cells(Tregs) and Th17 cells in the splenocytes from the treatment group with different doses of total glucosides of paeony(TGP). Methods The EAT model mice were randomly divided into 5 groups: model group, TGP high-dose group, TGP medium-dose group,TGP low-dose group and tripterygium glycosides group. Real-time PCR was performed to detect the expression of Foxp3 mRNA and RORγt mRNA in the splenocytes of each group. Flow cytometry was performed to detect the percentage of Treg and Th17 cells in the mice splenocytes of each group. Serum levels of TGF-β1 and IL-17 in each group were measured by ELISA. Serum level of TGAb in each group was detected by chemiluminescence assay.Results Foxp3 mRNA expression in the mice splenocytes in the medium-dose and in the high-dose TGP group was significantly higher than that in the model group(P< 0.05). The percentages of Treg cells in the medium-dose and the high-dose TGP group were significantly higher than that in the model group( P<0.01,P<0.05). The percentages of T cells secreting IL-17 in the low-, medium-and high-dose groups of TGP were significantly lower than that in the model group(P<0.001). Serum levels of TGF-β1 in the TGP medium-dose group and the TGP high-dose group were significantly higher thanthat in the model group( P<0.01).Serum levels of IL-17 in the TGP medium-dose group and the TGP high-dose group were significantly lower than that in the model group(P<0.01,P<0.05). Serum levels of TGAb in the TGP medium-dose group and the TGP high-dose group were significantly lower than that in the model group(P<0.05). Conclusions TGP can regulate the Treg/Th17 immune balance, which is potentially beneficial to the prognosis of EAT disease. The TGP medium-dose group and the TGP high-dose group have obvious curative effect.
引文
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[12] Korn T,Bettelli E,Oukka M,et al.IL-17 and Th17 Cells [J].Annu Rev Immunol,2009,27:485-517.
[2] Yu S,Sharp GC,Braley-Mullen H.Thyrocytes responding to IFN-gamma are essential for development of lymphocytic spontaneous autoimmune thyroiditis and inhibition of thyrocyte hyperplasia[J].J Immunol,2006,176:1259-1265.
[3] Koida S,Tsukasaki K,Tsuchiya T,et al.Primary T-cell lymphoma of the thyroid gland with chemokine receptors of Th1 phenotype complicating autoimmune thyroiditis[J].Haematologica,2007,92:37-40.
[4] 王萍,叶登美,窦德宇,等.白芍总苷对EAT小鼠Th1/Th2型细胞因子表达的影响[J].中国临床药理学与治疗学,2016,21:894-898.
[5] Zhao M,Liang GP,Tang MN,et al.Total glucosides of paeony induces regulatory CD4+ CD25+,T cells by increasing Foxp3,demethylation in lupus CD4+,T cells[J].Clin Immunol,2012,143:180-187.
[6] Lin J,Xiao L,Ouyang G,et al.Total glucosides of paeony inhibits Th1/Th17 cells via decreasing dendritic cells activation in rheumatoid arthritis[J].Cell Immunol,2012,280:156-163.
[7] Fontenot JD,Rasmussen JP,Williams LM,et al.Regulatory T cell lineage specification by the forkhead transcription factor Foxp3[J].Immunity,2005,22:329-341.
[8] Marie JC,Letterio JJ,Gavin M,et al.TGF-beta l maintains suppressor function and Foxp3 expression in CD4+CD25+regulatory T cells[J].J Exp Med,2005,201:1061-1067.
[9] Klatka M,Grywalska E,Partyka M,et al.Th17 and Treg cells in adolescents with Graves' disease.Impact of treatment with methimazole on these cell subsets[J].Autoimmunity,2014,47:201-211.
[10] Kristensen B,Hegedüs L,Madsen HO,et al.Altered balance between self-reactive Th17 cells and Th10 cells and between fulllength Foxp3 and Foxp3 splice variants in Hashimoto's thyroiditis[J].Clin Exp Immunol,2015,180:58-69.
[11] Huang B,Zhao J,Shen S,et al.Listeria monocytogenes promotes tumor growth via tumor cell toll-like receptor 2 signaling[J].Cancer Res,2007,67:4346-4352.
[12] Korn T,Bettelli E,Oukka M,et al.IL-17 and Th17 Cells [J].Annu Rev Immunol,2009,27:485-517.