雄激素对体外分化Th17细胞IL-17a、IL-17f、RORγt表达的影响
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摘要
目的研究雄激素对小鼠Th17细胞诱导分化模型中IL-17a、IL-17f、RORγt、mRNA表达的影响及其临床意义。方法提取小鼠脾脏淋巴细胞,采用免疫磁珠纯化CD4~+ CD62L~+T细胞,将其分为对照组、雄激素组、Th17分化组、雄激素组+Th17分化组。Th17分化条件采用对照组培养条件下加用anti-CD3ε、anti-CD28、antiIFN-γ、anti-IL-4抗体及IL-6、TGF-β1、IL-23等细胞因子培育,诱导原始T细胞向Th17细胞分化。雄激素+Th17分化组在Th17分化组的培养条件下加用雄激素。4组细胞经上述条件分别培养72h后,采用Realtime-PCR检测两组IL-17a、IL-17f、RORγt mRNA的表达验证Th17细胞分化的效果,探讨雄激素在Th17细胞分化中的作用。结果与对照组比较雄激素组IL-17a、IL-17f、RORγt mRNA增高,Th17分化组细胞经相关抗体及细胞因子刺激诱导分化后IL-17a、IL-17f、RORγt mRNA(P<0.01)表达明显增高,而雄激素+Th17分化组的IL-17a、IL-17f、RORγt mRNA表达最高(P<0.01)。结论雄激素可通过促进RORγt表达,增加IL-17a、IL-17f炎症因子释放起到促进炎症的作用。
Objective To research the influence of androgen on the expression of IL-17 a,IL-17f,RORγt mRNA in mice Th17 cell differentiation model,and its significance.Methods Mice spleen mononuclear lymphocytes were purified by mouse CD4~+ CD62L~+T cells isolation Kit Ⅱ. The CD4~+ CD62L~+T Cells were divided into control group,androgen group,Th17 group,Androgen + Th17 group. Control group cells were cultured with RPMI 1640,while the cells in Th17 differentiation group were cultured in RPMI 1640 with anti-mouse CD28,IL-6,TGF-β1,anti-mouse IL-4,anti-mouse IFN-γ,and mouse IL-23. The androgen + Th17 group were added androgen. All the cells were cultured at 37℃ 5% CO_2 for 72 hours. After harvesting,allthe cells IL-17 f,IL-17f,RORγt mRNA expression were detected by Real-time quantitative PCR for validation Th17 cell differentiation effect. Results Androgen group compared with controls,the expression of IL-17 a,IL-17f,RORγt mRNA were higher. The expression of IL-17f mRNA in Th17 cells differentiation group increase obviously( P < 0. 01),after stimulated by relevant antibodies and cytokines,while the expression of IL-17a,IL-17f,RORγt mRNA were upregulated in androgen + Th17 group( P < 0. 01). Conclusion Androgen can increase the expression of RORγt,IL-17a,IL-17f inflammatory factors and play an important role in promoting inflammation.
Objective To research the influence of androgen on the expression of IL-17 a,IL-17f,RORγt mRNA in mice Th17 cell differentiation model,and its significance.Methods Mice spleen mononuclear lymphocytes were purified by mouse CD4~+ CD62L~+T cells isolation Kit Ⅱ. The CD4~+ CD62L~+T Cells were divided into control group,androgen group,Th17 group,Androgen + Th17 group. Control group cells were cultured with RPMI 1640,while the cells in Th17 differentiation group were cultured in RPMI 1640 with anti-mouse CD28,IL-6,TGF-β1,anti-mouse IL-4,anti-mouse IFN-γ,and mouse IL-23. The androgen + Th17 group were added androgen. All the cells were cultured at 37℃ 5% CO_2 for 72 hours. After harvesting,allthe cells IL-17 f,IL-17f,RORγt mRNA expression were detected by Real-time quantitative PCR for validation Th17 cell differentiation effect. Results Androgen group compared with controls,the expression of IL-17 a,IL-17f,RORγt mRNA were higher. The expression of IL-17f mRNA in Th17 cells differentiation group increase obviously( P < 0. 01),after stimulated by relevant antibodies and cytokines,while the expression of IL-17a,IL-17f,RORγt mRNA were upregulated in androgen + Th17 group( P < 0. 01). Conclusion Androgen can increase the expression of RORγt,IL-17a,IL-17f inflammatory factors and play an important role in promoting inflammation.
引文
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[12]Yang XO,Pappu BP,Nurieva R,et al.T helper 17 lineage differentiation is programmed by orphan nuclear receptors ROR alpha and ROR gamma[J].Immunity,2008,28(1):29-39.
[2]任小丽,苏湘川,石青青,等.Th17细胞及白细胞介素17A、17F对特应性皮炎患者病情的影响[J].中国中西医结合皮肤性病学杂志,2016,15(3):163-165.
[3]盛国荣,严霞,刘海琴,等.复方利多卡因乳膏对激素依赖性皮炎Th17类细胞因子表达水平的影响[J].国医导报,2016,35(12):1312-6131.
[4]葛丽艳,田洁,汤新逸,等.痤疮丙酸杆菌通过促进Th1细胞和Th17细胞的分化诱发肉芽肿形成[J].细胞与分子免疫学杂志,2015,31(11):1447-1457.
[5]吴波,邵晓珊,吴文锦,等.青少年痤疮患者雄激素受体水平、雄激素受体基因CAG多态性的研究[J].贵州医药,2014,38(7):583-586.
[6]Korn T,Bettelli E,Oukka M,et al.IL-17 and Th17 Cells[J].Annu Rev Immunol,2009,27:485-517.
[7]Afzali B,Mitchell P,Lechler RI,et al.Translational minireview series on Th17 cells:induction of interleukin-17production by regulatory T cells[J].Clin Exp Immunol,2010,159(2):120-130.
[8]Hirahara K,Ghoreschi K,Laurence A,et al.Signal transduction pathways and transcriptional regulation in Th17 cell differentiation[J].Cytokine Growth Factor Rev,2010,21(6):425-434.
[9]陈嵘祎,周飞,周英,等.体外分化Th17细胞IL-17f,ctBP和Gfi1的表达[J].中国皮肤性病学杂志,2014,28(6):563-566.
[10]Rong-Yi Chen,Yi-Ming Fan,Qiuyang Zhang,et al.Estradiol inhibits Th17 cell differentiation through inhibition of RORg T transcription by recruiting the ERa/REA complex to estrogen response elements of the RORg T promoter[J].J Immunol,2015,194(8):4019-4028.
[11]马新华.舌丸治疗寻常性痤疮疗效观察及对血清IL-17的影响[J].新中医,2016,48(6):188-189.
[12]Yang XO,Pappu BP,Nurieva R,et al.T helper 17 lineage differentiation is programmed by orphan nuclear receptors ROR alpha and ROR gamma[J].Immunity,2008,28(1):29-39.