白介素-1β及NOD样受体蛋白3在痤疮炎性反应中的作用
|
摘要
目的:探讨痤疮的发病机制,为痤疮的非抗生素治疗提供新思路。方法:分别使用感染复数(multiplicity ofinfection,MOI)为0,10,20,30的痤疮丙酸杆菌菌悬液刺激正常人表皮角质形成细胞(normalhumanepidermal keratinocyte,NHEK)12,24,36h后,使用ELISA,real-timePCR检测NHEK中IL-1β的蛋白及mRNA的表达。另设3组:空白对照组,未进行任何预处理的NHEK;阳性对照组,仅使用MOI为30的痤疮丙酸杆菌菌悬液刺激NHEK;siRNA组,使用特异性小干扰RNA(siRNA)预处理以后再使用MOI为30的痤疮丙酸杆菌菌悬液刺激NHEK,使用蛋白质印迹法检测3组中NOD样受体蛋白3(nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3,NLRP3)炎性小体的表达情况,使用ELISA,real-time PCR检测3组NHEK中IL-1β蛋白及mRNA的表达。结果:不同浓度痤疮丙酸杆菌菌悬液刺激NHEK后,IL-1β表达量较空白对照组均有不同程度的升高,升高程度呈时间及剂量相关性(P<0.05)。siRNA预处理NHEK后,NLRP3炎性小体以及IL-1β的表达量明显降低,但仍高于空白对照组(P<0.05)。结论:痤疮丙酸杆菌刺激NHEK分泌IL-1β可能需要NLRP3炎性小体的参与;可通过抑制NLRP3炎性小体的活性来抑制由痤疮丙酸杆菌导致的炎性反应,从而减少抗生素的使用。
Objective:To investigate the pathogenesis of acne vulgaris,and to provide new ideas for nonantibiotic therapy for acne vulgaris.Methods:Normal human epidermal keratinocyte(NHEK) was exposed to Propionibacterium acnes(P.acnes) [multiplicity of infection(MOI)=10,20,30] for 12,24,or 36 hours.The enzyme-linked immunosorbent assay(ELISA) and real-time PCR were used to detect the protein and mRNA of IL-1β in NHEK.Three groups were set up as follows:A negative control group(no NHEK pretreatment),a positive control group(P.acnes was used to stimulate NHEK),and a siRNA group(pretreated NHEK with siRNA).ELISA,real-time PCR,and Western blotting were used to detect the protein,mRNA of IL-1β and nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3(NLRP3) in NHEK.Results:IL-1β of NHEK in the positive control group was significantly increased in a time and dose-dependent manner compared with the negative control group(P<0.05).After pretreating NHEK with siRNA,IL-1β level was decreased compared with the positive control group,but it was higher than that in the negative control group(P<0.05).Conclusion:P.ances can stimulate NHEK to secrete IL-1β,and the process is possibly involved in NLRP3.The inflammatory response induced by P.ances could be inhibited by suppressing the activity of NLRP3.
Objective:To investigate the pathogenesis of acne vulgaris,and to provide new ideas for nonantibiotic therapy for acne vulgaris.Methods:Normal human epidermal keratinocyte(NHEK) was exposed to Propionibacterium acnes(P.acnes) [multiplicity of infection(MOI)=10,20,30] for 12,24,or 36 hours.The enzyme-linked immunosorbent assay(ELISA) and real-time PCR were used to detect the protein and mRNA of IL-1β in NHEK.Three groups were set up as follows:A negative control group(no NHEK pretreatment),a positive control group(P.acnes was used to stimulate NHEK),and a siRNA group(pretreated NHEK with siRNA).ELISA,real-time PCR,and Western blotting were used to detect the protein,mRNA of IL-1β and nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3(NLRP3) in NHEK.Results:IL-1β of NHEK in the positive control group was significantly increased in a time and dose-dependent manner compared with the negative control group(P<0.05).After pretreating NHEK with siRNA,IL-1β level was decreased compared with the positive control group,but it was higher than that in the negative control group(P<0.05).Conclusion:P.ances can stimulate NHEK to secrete IL-1β,and the process is possibly involved in NLRP3.The inflammatory response induced by P.ances could be inhibited by suppressing the activity of NLRP3.
引文
[1]Szabo K,Erdei L,Bolla BS,et al.Factors shaping the composition of the cutaneous microbiota[J].Br J Dermatol,2017,176(2):344-351.
[2]Sanford JA,Gallo RL.Functions of the skin microbiota in health and disease[J].Semin Immunol,2013,25(5):370-377.
[3]Qin M,Pirouz A,Kim MH,et al.Propionibacterium acnes induces IL-1βsecretion via the NLRP3 inflammasome in human monocytes[J]Dermatology,2014,134(2):381-388.
[4]Askari N,Ghazanfari T,Yaraee R,et al.Association between acne and serum pro-inflammatory cytokines(IL-1α,IL-1β,IL-1Ra,IL-6IL-8,IL-12)in mustard gas-exposed patients:sardasht-iran cohort study[J].Arch Iran Med,2017,20(2):86-91.
[5]Dai X,Sayama K,Tohyama M,et al.Mite allergen is a danger signal for the skin via activation of inflammasome in keratinocytes[J].JAllergy Clin Immun,2011,127(3):1-4.
[6]Shaheen B,Gonzalez M.Acne sans P.acnes[J].J Europ Acad Dermatol Venereol,2013,27(1):1-10.
[7]Dessinioti C,Katsambas A.Propionibacterium acnes and antimicrobial resistance in acne[J].Clin Dermatol,2017,35(2):163-167.
[8]Contassot E,French LE.New insights into acne pathogenesis:Propionibacterium acnes activates the inflammasome[J].J Invest Dermatol,2014,134(2):310-313.
[9]Berolia S,Evasa R,Fredrik E,et al.Propionibacterium acnes activates caspase-1 in human neutrophils[J].APMIS,2013,121(7):652-663.
[10]林新瑜,罗旭松,董巍,等.痤疮患者血清IL-1α,IL-6,IL-8和TNF-α水平的检测分析[J].临床皮肤科杂志,2003,32(6):335-336.LIN Xinyu,LUO Xusong,DONG Wei,et al.Serum levels of interleukin-1α,interleukin-6,interleukin-8 and tumour necrosis factor-αin acne patients[J].Journal of Clinical Dermatology,200332(6):335-336.
[11]Beylot C,Auffret N,Poli F,et al.Propionibacterium acnes:an update on its role in the pathogenesis of acne[J].J Eur Acad Dermatol Venereol,2014,28(3):271-278.
[12]Yang G,Yeon SH,L ee H E,et al.Suppression of NLR P3inflammasome by oral treatment with sulforaphane alleviates acute gouty inflammation[J].Rheumatology,2018,57(4):1-10.
[13]Jiang TC,Jiang DL,Zhang LR,et al.Anagliptin ameliorates high glucose-induced endothelial dysfunction via suppression of NLRP3inflammasome activation mediated by SIRT1[J].Mol Immunol2019,107(9):54-60.
[2]Sanford JA,Gallo RL.Functions of the skin microbiota in health and disease[J].Semin Immunol,2013,25(5):370-377.
[3]Qin M,Pirouz A,Kim MH,et al.Propionibacterium acnes induces IL-1βsecretion via the NLRP3 inflammasome in human monocytes[J]Dermatology,2014,134(2):381-388.
[4]Askari N,Ghazanfari T,Yaraee R,et al.Association between acne and serum pro-inflammatory cytokines(IL-1α,IL-1β,IL-1Ra,IL-6IL-8,IL-12)in mustard gas-exposed patients:sardasht-iran cohort study[J].Arch Iran Med,2017,20(2):86-91.
[5]Dai X,Sayama K,Tohyama M,et al.Mite allergen is a danger signal for the skin via activation of inflammasome in keratinocytes[J].JAllergy Clin Immun,2011,127(3):1-4.
[6]Shaheen B,Gonzalez M.Acne sans P.acnes[J].J Europ Acad Dermatol Venereol,2013,27(1):1-10.
[7]Dessinioti C,Katsambas A.Propionibacterium acnes and antimicrobial resistance in acne[J].Clin Dermatol,2017,35(2):163-167.
[8]Contassot E,French LE.New insights into acne pathogenesis:Propionibacterium acnes activates the inflammasome[J].J Invest Dermatol,2014,134(2):310-313.
[9]Berolia S,Evasa R,Fredrik E,et al.Propionibacterium acnes activates caspase-1 in human neutrophils[J].APMIS,2013,121(7):652-663.
[10]林新瑜,罗旭松,董巍,等.痤疮患者血清IL-1α,IL-6,IL-8和TNF-α水平的检测分析[J].临床皮肤科杂志,2003,32(6):335-336.LIN Xinyu,LUO Xusong,DONG Wei,et al.Serum levels of interleukin-1α,interleukin-6,interleukin-8 and tumour necrosis factor-αin acne patients[J].Journal of Clinical Dermatology,200332(6):335-336.
[11]Beylot C,Auffret N,Poli F,et al.Propionibacterium acnes:an update on its role in the pathogenesis of acne[J].J Eur Acad Dermatol Venereol,2014,28(3):271-278.
[12]Yang G,Yeon SH,L ee H E,et al.Suppression of NLR P3inflammasome by oral treatment with sulforaphane alleviates acute gouty inflammation[J].Rheumatology,2018,57(4):1-10.
[13]Jiang TC,Jiang DL,Zhang LR,et al.Anagliptin ameliorates high glucose-induced endothelial dysfunction via suppression of NLRP3inflammasome activation mediated by SIRT1[J].Mol Immunol2019,107(9):54-60.