麻黄细辛附子汤纠正TSLP刺激树突状细胞后Th1/Th2偏移研究
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摘要
目的研究麻黄细辛附子汤对免疫应答启动分子胸腺基质淋巴生成素(thymic stromal lymphopoietin,TSLP)刺激树突状细胞后对Th1/Th2失衡的影响,探究麻黄细辛附子汤治疗过敏性鼻炎可能的作用靶点及药理机制。方法体外诱导培养小鼠骨髓来源的树突状细胞,7天培养成功后,设空白组、TSLP组、麻黄细辛附子汤组。TSLP组与麻黄细辛附子汤组的树突状细胞均用TSLP刺激,麻黄细辛附子汤组同时用麻黄细辛附子汤进行干预,24小时后收集细胞,检测IL-4 mRNA、IFN-γ mRNA表达水平;收集实验各组细胞上清,检测细胞因子IL-13、IFN-γ的含量。结果 (1)与空白组比较,TSLP组IL-4 m RNA表达显著增多(P<0.05),而麻黄细辛附子汤组表达显著减少(P<0.05),麻黄细辛附子汤组IL-4 mRNA表达较TSLP组显著减少(P<0.01);(2)与空白组比较,麻黄细辛附子汤组及TSLP组IFN-γ mRNA表达较均显著增多(P<0.05),且麻黄细辛附子汤组较TSLP组升高更明显(P<0.01);(3)与空白组比较,TSLP组IFN-γ m RNA/IL-4 mRNA无明显差异(P>0.05),而麻黄细辛附子汤组较TSLP干预组与空白组显著升高(P<0.01);(4)与空白组比较,TSLP组分泌IL-13显著增多(P<0.05),而IFN-γ显著减少(P<0.01),麻黄细辛附子汤组分泌IL-13含量较TSLP干预组及空白组均显著减少(P<0.05),麻黄细辛附子汤组IFN-γ含量较空白组无差异(P>0.05),与TSLP组相比显著增多(P<0.01)。结论麻黄细辛附子汤汤能上调TSLP刺激树突状细胞后IFN-γm RNA的表达,下调IL-4 mRNA,升高IFN-γ mRNA/IL-4 mRNA比值并抑制TSLP刺后激树突状细胞分泌IL-13,促进IFN-γ的分泌,对T细胞Th2分化进行调控,恢复Th1/Th2失衡,可对过敏性疾病发挥治疗作用。
Objective To study on the effect of Mahuang Xixin Fuzi decoction on the imbalance of Th1/Th2 about dendritic cells stimulated by TSLP,and to explore the potential targets and pharmacological mechanisms of Mahuang Xixin Fuzi decoction to cure Allergic Rhinitis( AR). Methods The dendritic cells derived from bone marrow of mice were induced in vitro. After 7 days of successful culture,blank group,TSLP group,and Mahuang Xixin Fuzi decoction group were set up. Dendritic cells from TSLP group and Mahuang Xixin Fuzi decoction group were stimulated by TSLP. Mahuang Xixin Fuzi decoction group were intervened with Mahuang Xixin Fuzi decoction,and the cells were collected. After 24 hours,the cells were collected for detecting IL-14 mRNA and IFN-γ m RNA,and the contents of IL-13 and IFN-γ in DCs supernatants were measured. Results( 1) Compared with the blank group,the expression of IL-4 mRNA in TSLP group was increased significantly( P< 0.05),while the expression of IL-4 mRNA in Mahuang Xixin Fuzi Decoction group was decreased significantly( P< 0. 05). The expression of IL-4 mRNA in Mahuang Xixin Fuzi decoction group was significantly less than TSLP group( P< 0. 01);( 2) Compared with the blank group,the expression of IFN-γ mRNA of TSLP group and Mahuang Xixin Fuzi decoction group was significantly increased( P < 0. 05),and the expression of IFN-γ mRNA in Mahuang Xixin Fuzi decoction group were more than TSLP group( P < 0. 01);( 3) Compared with the blank group,the IFN-γ m RNA/IL-4 mRNA in TSLP group had no difference( P> 0.05),while the ratio was raised in Mahuang Xixin Fuzi decoction group( P< 0.01);( 4) Compared with the blank group,the content of IL-13 in TSLP group was increased significantly( P < 0. 05),while the content of IFN-γ was lower( P< 0. 01); the content of IL-13 in Mahuang Xixin Fuzi decoction group was lower than that in TSLP group( P< 0.01) and the bank group( P< 0. 05); the content of IFN-γ in Mahuang Xixin Fuzi decoction group had no difference compared with the bank group( P > 0. 05),and the content of IFN-γ in Mahuang Xixin Fuzi decoction group was increased than TSLP group( P< 0.01). Conclusion Mahuang Xixin Fuzi decoction can upregulate the expression of IFN-γ mRNA and down-regulate the expression of IL-4 mRNA of DCs stimulated by TSLP,can raise the ratio of IFN-γ m RNA/IL-4 m RNA. It also inhibits the secretion of IL-13 and promotes the secretion of IFN-γ,regulates the imbalance of Th1/Th2 and plays an important role in allergic diseases.
Objective To study on the effect of Mahuang Xixin Fuzi decoction on the imbalance of Th1/Th2 about dendritic cells stimulated by TSLP,and to explore the potential targets and pharmacological mechanisms of Mahuang Xixin Fuzi decoction to cure Allergic Rhinitis( AR). Methods The dendritic cells derived from bone marrow of mice were induced in vitro. After 7 days of successful culture,blank group,TSLP group,and Mahuang Xixin Fuzi decoction group were set up. Dendritic cells from TSLP group and Mahuang Xixin Fuzi decoction group were stimulated by TSLP. Mahuang Xixin Fuzi decoction group were intervened with Mahuang Xixin Fuzi decoction,and the cells were collected. After 24 hours,the cells were collected for detecting IL-14 mRNA and IFN-γ m RNA,and the contents of IL-13 and IFN-γ in DCs supernatants were measured. Results( 1) Compared with the blank group,the expression of IL-4 mRNA in TSLP group was increased significantly( P< 0.05),while the expression of IL-4 mRNA in Mahuang Xixin Fuzi Decoction group was decreased significantly( P< 0. 05). The expression of IL-4 mRNA in Mahuang Xixin Fuzi decoction group was significantly less than TSLP group( P< 0. 01);( 2) Compared with the blank group,the expression of IFN-γ mRNA of TSLP group and Mahuang Xixin Fuzi decoction group was significantly increased( P < 0. 05),and the expression of IFN-γ mRNA in Mahuang Xixin Fuzi decoction group were more than TSLP group( P < 0. 01);( 3) Compared with the blank group,the IFN-γ m RNA/IL-4 mRNA in TSLP group had no difference( P> 0.05),while the ratio was raised in Mahuang Xixin Fuzi decoction group( P< 0.01);( 4) Compared with the blank group,the content of IL-13 in TSLP group was increased significantly( P < 0. 05),while the content of IFN-γ was lower( P< 0. 01); the content of IL-13 in Mahuang Xixin Fuzi decoction group was lower than that in TSLP group( P< 0.01) and the bank group( P< 0. 05); the content of IFN-γ in Mahuang Xixin Fuzi decoction group had no difference compared with the bank group( P > 0. 05),and the content of IFN-γ in Mahuang Xixin Fuzi decoction group was increased than TSLP group( P< 0.01). Conclusion Mahuang Xixin Fuzi decoction can upregulate the expression of IFN-γ mRNA and down-regulate the expression of IL-4 mRNA of DCs stimulated by TSLP,can raise the ratio of IFN-γ m RNA/IL-4 m RNA. It also inhibits the secretion of IL-13 and promotes the secretion of IFN-γ,regulates the imbalance of Th1/Th2 and plays an important role in allergic diseases.
引文
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[11]Liu YJ.THymic stromal lymphopoietin and OX40 ligand pa THway in THeinitiation of dendritic cell-mediated allergic inflammation[J].J Allergy Clin Immunol,2007,120(2):238-244.
[12]Cianferoni Al,Spergel J.THe importance of TSLP in allergic disease and its role as a potential THerapeutic target[J].Expert Rev Clin Immunol,2014,10(11):1463-1474.
[13]Divekar R,Kita H.Recent advances in epi THelium-derived cytokines(IL-33,IL-25,and THymic stromal lymphopoietin)and allergic inflammation[J].Curr Opin Allergy Clin Immunol,2015,15(1):98-103.
[14]Kaur D,Brightling C.OX40/OX40 ligand interactions in T-cell regulation and asthma.[J].Chest,2012,141(2):494-499.
[15]Leaker B R,Malkov V A,Mogg R,et al.The nasal mucosal late allergic reaction to grass pollen involves type 2 inflammation(IL-5 and IL-13),the inflammasome(IL-1β),and complement[J].Mucosal Immunology,2016,10(2):408.
[16]Kutukculer N,Azarsiz E,Aksu G,et al.Cd4+Cd25+Foxp3+Tregulatory cells,Th1(Ccr5,Il-2,Ifn-Γ)and Th2(Ccr4,Il-4,Il-13)type chemokine receptors and intracellular cytokines in children with common variable immunodeficiency[J].International Journal of Immunopathology&Pharmacology,2015,29(2):241.
[17]Mertens T C J,Hiemstra P S,Taube C.Azithromycin differentially affects the IL-13 induced expression profile in human bronchial epithelial cells[J].Pulmonary Pharmacology&Therapeutics,2016,39(5):14-20.
[18]王芳,胡晓艳.玉屏风散对变应性鼻炎模型大鼠血清Ig E、IL-4和IFN-r含量的影响[J].中国中西医结合耳鼻咽喉科杂志,2016,24(2):81-83.
[19]乔占清,张俊.麻黄连翘赤小豆汤加减对肺经郁热型变应性鼻炎Th1/Th2细胞,Th17细胞和调节性T细胞因子的影响[J].中国实验方剂学杂志,2016,(11):165-169.
[2]Kakli H A,Riley T D.Allergic Rhinitis[J].Primary Care,2016,43(3):465-475.
[3]Lambrecht B N,Hammad H.Function of Dendritic Cell Subsets in Allergic Disease[M]//Allergy Frontiers:Classification and Pathomechanisms.2009:209-230.
[4]Cao PP,Shi LL,Xu K,et al.Dendritic cells in inflammatory sinonasal diseases[J].Clinical&Experimental Allergy Journal of the British Society for Allergy&Clinical Immunology,2016,46(7):894-906.
[5]Ying S,O’Connor B,Ratoff J,et al.THymic stromal lym-phopoietin expression is increased in as THmatic airways andcorrelates wi TH expression of TH2-attracting chemokines anddisease severity[J].J Immunol,2005,174(12):8183-8190.
[6]Soumelis V,Reche PA,Kanzler H,et al.Human epi THelialcells trigger dendritic cell mediated allergic inflammation byproducing TSLP[J].Nat Immunol,2002,3(7):673-680.
[7]Akasaki S,Matsushita K,Kato Y,et al.Murine allergic rhinitis and nasal Th2 activation are mediated via TSLP-and IL-33-signaling pathways[J].International Immunology,2016,28(2):65.
[8]Yong M K.Prolonged Allergen Exposure Causes TSLP-Mediated Th2-Skewing in Mouse Models of Chronic Rhinosinusitis[J].Retour Au Numéro,2016,137(2):67.
[9]Inaba K,Inaba M,Romani N,et al.Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor[J].Journal of Experimental Medicine,1992,176(6):1693-1702.
[10]田晋生,邓勇志.IL-4对小鼠骨髓树突状细胞表面分子CD11c、CD80、CD86表达的影响及其意义[J].中华临床医师杂志(电子版),2014,(5):74-76.
[11]Liu YJ.THymic stromal lymphopoietin and OX40 ligand pa THway in THeinitiation of dendritic cell-mediated allergic inflammation[J].J Allergy Clin Immunol,2007,120(2):238-244.
[12]Cianferoni Al,Spergel J.THe importance of TSLP in allergic disease and its role as a potential THerapeutic target[J].Expert Rev Clin Immunol,2014,10(11):1463-1474.
[13]Divekar R,Kita H.Recent advances in epi THelium-derived cytokines(IL-33,IL-25,and THymic stromal lymphopoietin)and allergic inflammation[J].Curr Opin Allergy Clin Immunol,2015,15(1):98-103.
[14]Kaur D,Brightling C.OX40/OX40 ligand interactions in T-cell regulation and asthma.[J].Chest,2012,141(2):494-499.
[15]Leaker B R,Malkov V A,Mogg R,et al.The nasal mucosal late allergic reaction to grass pollen involves type 2 inflammation(IL-5 and IL-13),the inflammasome(IL-1β),and complement[J].Mucosal Immunology,2016,10(2):408.
[16]Kutukculer N,Azarsiz E,Aksu G,et al.Cd4+Cd25+Foxp3+Tregulatory cells,Th1(Ccr5,Il-2,Ifn-Γ)and Th2(Ccr4,Il-4,Il-13)type chemokine receptors and intracellular cytokines in children with common variable immunodeficiency[J].International Journal of Immunopathology&Pharmacology,2015,29(2):241.
[17]Mertens T C J,Hiemstra P S,Taube C.Azithromycin differentially affects the IL-13 induced expression profile in human bronchial epithelial cells[J].Pulmonary Pharmacology&Therapeutics,2016,39(5):14-20.
[18]王芳,胡晓艳.玉屏风散对变应性鼻炎模型大鼠血清Ig E、IL-4和IFN-r含量的影响[J].中国中西医结合耳鼻咽喉科杂志,2016,24(2):81-83.
[19]乔占清,张俊.麻黄连翘赤小豆汤加减对肺经郁热型变应性鼻炎Th1/Th2细胞,Th17细胞和调节性T细胞因子的影响[J].中国实验方剂学杂志,2016,(11):165-169.