加味过敏煎对过敏性哮喘小鼠Treg细胞特异性转录因子的影响
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摘要
目的通过研究加味过敏煎对Treg转录因子的影响,为加味过敏煎在哮喘免疫治疗中的机制及临床应用提供理论与实验依据。方法将60只BALB/c雌性小鼠随机分为6组,即空白组,模型组,地塞米松组(1.25 mg·kg~(-1)),加味过敏煎高、中、低剂量组(含生药量分别为54.4,27.2,13.6 g·kg~(-1)),每组10只。采用卵白蛋白(OVA)致敏和激发构建小鼠哮喘模型。采用实时荧光定量PCR法和WB法检测小鼠肺组织中Treg细胞特异性转录因子Foxp3的mRNA与蛋白表达水平。结果实时荧光定量PCR结果显示,与模型组比较,加味过敏煎高、中、低剂量组均可升高Foxp3的mRNA表达水平(P<0.05,P<0.01)。WB结果显示,与模型组比较,加味过敏煎高、中剂量组对Foxp3的蛋白表达则有不同程度的提高,但差异无统计学意义(P>0.05)。结论加味过敏煎治疗过敏性哮喘的免疫机制可能是上调Treg细胞特异性转录因子Foxp3的基因表达。
Objective To investigate the effects of modified Guomin Decoction(MGD)on the Treg cell transcription factor,thus to provide theoretical and experimental evidence for the clinical application and therapeutic mechanism of MGD in asthma immunotherapy.Methods Sixty female SPF BALB/c mice aged 4-5 weeks were randomly and evenly divided into 6 groups,blank control group,model control group,dexamethasone(DXM)positive control group(1.25mg·kg~(-1)),and high-,medium-and low-dose MGD groups(containing crude drug 54.4,27.2,13.6 g·kg~(-1),respectively).Mouse asthma model was established by sensitation and stimulation with ovalbumin(OVA).The real-time fluorescence quantitative PCR and the Western blot method were used to determine the mRNA and protein expression levels of pulmonary Foxp3,which is a Treg cell specific transcription factor.Results The outcome of the real-time fluorescence quantitative PCR showed that the high,medium and low dose of MGD could increase Foxp3 mRNA expression level as compared with the model control group(P<0.05,P<0.01).The results of Western blot medthod indicated that Foxp3 protein expression level was increased to various extents in high-and medium-dose MGD groups as compared with the model control group,but the differences were insignificant(P>0.05).Conclusion The immunotherapeutic effect of MGD against allergic asthma may be achieved by the up-regulation of the gene expression of the Treg cell specific transcription factor Foxp3.
Objective To investigate the effects of modified Guomin Decoction(MGD)on the Treg cell transcription factor,thus to provide theoretical and experimental evidence for the clinical application and therapeutic mechanism of MGD in asthma immunotherapy.Methods Sixty female SPF BALB/c mice aged 4-5 weeks were randomly and evenly divided into 6 groups,blank control group,model control group,dexamethasone(DXM)positive control group(1.25mg·kg~(-1)),and high-,medium-and low-dose MGD groups(containing crude drug 54.4,27.2,13.6 g·kg~(-1),respectively).Mouse asthma model was established by sensitation and stimulation with ovalbumin(OVA).The real-time fluorescence quantitative PCR and the Western blot method were used to determine the mRNA and protein expression levels of pulmonary Foxp3,which is a Treg cell specific transcription factor.Results The outcome of the real-time fluorescence quantitative PCR showed that the high,medium and low dose of MGD could increase Foxp3 mRNA expression level as compared with the model control group(P<0.05,P<0.01).The results of Western blot medthod indicated that Foxp3 protein expression level was increased to various extents in high-and medium-dose MGD groups as compared with the model control group,but the differences were insignificant(P>0.05).Conclusion The immunotherapeutic effect of MGD against allergic asthma may be achieved by the up-regulation of the gene expression of the Treg cell specific transcription factor Foxp3.
引文
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[12]LU C H,WU C J,CHAN C C,et al.DNA Methyltransferase Inhibitor Promotes Human CD4+CD25h FOXP3+Regulatory T Lymphocyte Induction under Suboptimal TCR Stimulation[J].Front Immunol,2016,8(7):488.
[13]PARK S,CHUNG H S,SHIN D,et al.Adenovirus-mediated Foxp3expression in lung epithelial cells reduces airway inflammation in ovalbumin and cockroach-induced asthma model[J].Exp Mol Med,2016,48(9):259.
[2]胡斯明,罗雅玲,赖文岩,等.调节性T细胞/Th17在支气管哮喘小鼠气道炎症过程中的变化[J].中国现代医学杂志,2009,19(19):2881-2884.
[3]卢世秀.晁恩祥肺系疾病治疗特色[J].中国中医药信息杂志,2007,14(2):75-76.
[4]孙静瑛.从过敏煎的运用谈辨病用药与辨证用药[J].吉林中医药,2004,24(4):45.
[5]李彦军,龚盟,马淑然,等.过敏煎对SD大鼠血中Ig E变化影响的实验研究[J].辽宁中医杂志,2010,37(1):177-178.
[6]覃骊兰,马淑然.加味过敏煎对哮喘小鼠气道炎症抑制作用的实验研究[J].广州中医药大学学报,2014,31(2):275-279.
[7]PALA P,MESSAGE S D,JOHNSTON S L,et al.Increased aeroallergen-specific interleukin-4-producting T cells in asthmatic adults[J].Clin Exp Allergy,2002,32(12):1739-1744.
[8]WAKASHIN H,HIROSE K,MAEZAWA Y,et al.IL-23 and Th17cells enhance Th12-cell-mediated eosinophilic airway inflammation in mice[J].Am J Respir Crit Care Med,2008,178(10):1023-1032.
[9]ZHANH J G,CHEN X J,LIU T,et al.FOXP3+associated with the pro-inflammatory regulatory T and T helper 17 effector cells in asthma patients[J].Exp Ther Med,2016,12(4):2753-2758.
[10]WANG Y H,LIU Y J.The IL-17 cytokine family and their role in allergic inflammation[J].Curr Opin Immunol,2008,20(6):697-702.
[11]USO M,JANTUS L E,GALLACH S,et al.Association of Foxp3expression with prognosis in resected non-small cell lung cancer(NSCLC)[J].J Clin Oncol.,2011,29(15):21114.
[12]LU C H,WU C J,CHAN C C,et al.DNA Methyltransferase Inhibitor Promotes Human CD4+CD25h FOXP3+Regulatory T Lymphocyte Induction under Suboptimal TCR Stimulation[J].Front Immunol,2016,8(7):488.
[13]PARK S,CHUNG H S,SHIN D,et al.Adenovirus-mediated Foxp3expression in lung epithelial cells reduces airway inflammation in ovalbumin and cockroach-induced asthma model[J].Exp Mol Med,2016,48(9):259.