葛根素对小鼠皮肤来源朗格汉斯细胞成熟分化及炎症因子分泌的影响
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摘要
目的:观察葛根素(puerarin)对小鼠皮肤来源朗格汉斯细胞数量、成熟及炎症因子分泌的影响,以期探讨葛根素在炎症相关性疾病治疗中的潜在机制。方法:葛根素作用于体外培养的C57BL/6小鼠皮肤朗格汉斯细胞后,使用流式细胞术检测葛根素对朗格汉斯细胞数量、抗原捕获、MHC-Ⅱ表达等成熟能力的影响;同时利用小鼠细胞因子抗体芯片技术检测培养上清中细胞因子的变化。结果:葛根素作用后,小鼠朗格汉斯细胞数量无明显变化,但抗原捕获能力显著增加;MHC-Ⅱ~(high)亚群细胞比例降低,MHC-Ⅱ~(low)亚群细胞比例则增多;MHC-Ⅱ~+CD80~+和MHC-Ⅱ~+CD86~+细胞比例显著降低;培养上清中IL-1β、MCP-1的分泌也受到明显抑制。结论:葛根素抑制朗格汉斯细胞成熟能力和炎症因子分泌水平,有助于阐明朗格汉斯细胞在炎症相关性疾病中的作用机制具有重要临床意义。
OBJECTIVE To observe the effect of puerarin on the secretion of Langerhans cells from mouse skin and the secretion of inflammatory cytokines,and to investigate the potential immunological mechanism of puerarin for inflammation-relat-ed diseases.METHODS After puerarin was applied to the mouse Langerhans cell,the effects of puerarin on the cell number,antigen capture and MHC-Ⅱexpression were detected by flow cytometry.The mouse cytokine antibody chip technology wasused to detect the changes in culture supernatant cytokines.RESULTS After puerarin treatment,the number of mouse Langer-hans cells did not significantly change.However,the capacity of antigen capture was increased.Nevertheless,the proportionof MHC-Ⅱ~(high) subpopulation was decreased,while the proportion of MHC-Ⅱ~(low)subpopulation was increased.The proportionsof MHC-Ⅱ~+CD80~+and MHC-Ⅱ~+CD86~+cells were significantly decreased.The secretions of IL-1βand MCP-1 in the culturesupernatant were also significantly inhibited.CONCLUSIONPuerarin inhibits Langerhans cell maturation and inflammatory cy-tokine secretion,indicating it has important clinical significance for elucidating the mechanism of Langerhans cells in inflammation-related diseases.
OBJECTIVE To observe the effect of puerarin on the secretion of Langerhans cells from mouse skin and the secretion of inflammatory cytokines,and to investigate the potential immunological mechanism of puerarin for inflammation-relat-ed diseases.METHODS After puerarin was applied to the mouse Langerhans cell,the effects of puerarin on the cell number,antigen capture and MHC-Ⅱexpression were detected by flow cytometry.The mouse cytokine antibody chip technology wasused to detect the changes in culture supernatant cytokines.RESULTS After puerarin treatment,the number of mouse Langer-hans cells did not significantly change.However,the capacity of antigen capture was increased.Nevertheless,the proportionof MHC-Ⅱ~(high) subpopulation was decreased,while the proportion of MHC-Ⅱ~(low)subpopulation was increased.The proportionsof MHC-Ⅱ~+CD80~+and MHC-Ⅱ~+CD86~+cells were significantly decreased.The secretions of IL-1βand MCP-1 in the culturesupernatant were also significantly inhibited.CONCLUSIONPuerarin inhibits Langerhans cell maturation and inflammatory cy-tokine secretion,indicating it has important clinical significance for elucidating the mechanism of Langerhans cells in inflammation-related diseases.
引文
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[16]Zhou J,Fang SP,Qi Y,et al.Effects of Gegen Decoction on Inflammatory Mediators in Synovial Fluid of Rats with Adjuvant Arthritis[J].Chin J Exp Tradit Med Formul(中国实验方剂学杂志),2001,3:29-31.
[2]Gao Z,Wei B,Qian C.Puerarin injection for treatment of unstable angina pectoris:a meta-analysis and systematic review[J].Int J Clin Exp Med,2015,8(9):14577-14594.
[3]Lee JH,Jeon YD,Lee YM,et al.The suppressive effect of puerarin on atopic dermatitis-like skin lesions through regulation of inflammatory mediators in vitro and in vivo[J].Biochem Biophys Res Commun,2018,498(4):707-714.
[4]Wang C,Wang W,Jin X,et al.Puerarin attenuates inflammation and oxidation in mice with collagen antibody-induced arthritis via TLR4/NF-kappaB signaling[J].Mol Med Rep,2016,14(2):1365-1370.
[5]Arsovic N,Tomanovic N and Bukurov B.Sudden sensorineural hearing loss as first presenting symptom of unifocal Langerhans cell histiocytosis in the temporal bone[J].Otol Neurotol,2013,34(3):e24-25.
[6]Suzuki H,Nishizawa S,Hohchi N,et al.Langerhans cell histiocytosis of the petrous bone with sudden sensorineural hearing loss[J].Neurol Med-Chir(Tokyo),2010,50(8):693-697.
[7]Yan LP,Zhuang YL,Chan SW,et al.Analysis of the mechanisms underlying the endothelium-dependent antivasoconstriction of puerarin in rat aorta[J].Naunyn-Schmiedeberg′s Arch Pharmacol,2009,379(6):587-597.
[8]Kashem SW,Kaplan DH.Isolation of Murine Skin Resident and Migratory Dendritic Cells via Enzymatic Digestion[J].Cur Protoc Immunol,2018,121(1):e45.
[9]Wei SY,Chen Y,Xu XY.Progress on the pharmacological research of puerarin:a review[J].Chin J Nat Med,2014,12(6):407-414.
[10]Cronkite DA,Strutt TM.The Regulation of Inflammation by Innate and Adaptive Lymphocytes[J].J Immunol Res,2018,2018:1467538.
[11]Kassner SS,Schottler S,Bonaterra GA,et al.Proinflammatory and proadhesive activation of lymphocytes and macrophages in sudden sensorineural hearing loss[J].Audiol Neuro-otol,2011,16(4):254-262.
[12]Mbongue JC,Nieves HA,Torrez TW,et al.The Role of Dendritic Cell Maturation in the Induction of Insulin-Dependent Diabetes Mellitus[J].Front Immunol,2017,8:327.
[13]Mohamadzadeh M,Poltorak AN,Bergstressor PR,et al.Dendritic cells produce macrophage inflammatory protein-1gamma,a new member of the CC chemokine family[J].J Immunol,1996,156(9):3102-3106.
[14]Nakamura K,Williams IR and Kupper TS.Keratinocyte-derived monocyte chemoattractant protein 1(MCP-1):analysis in a transgenic model demonstrates MCP-1can recruit dendritic and Langerhans cells to skin[J].J Investi Dermatol,1995,105(5):635-643.
[15]Tao Z,Ge Y,Zhou N,et al.Puerarin inhibits cardiac fibrosis via monocyte chemoattractant protein(MCP)-1and the transforming growth factor-beta1(TGF-beta1)pathway in myocardial infarction mice[J].Am J Transl Res,2016,8(10):4425-4433.
[16]Zhou J,Fang SP,Qi Y,et al.Effects of Gegen Decoction on Inflammatory Mediators in Synovial Fluid of Rats with Adjuvant Arthritis[J].Chin J Exp Tradit Med Formul(中国实验方剂学杂志),2001,3:29-31.