SEB对人HaCaT细胞株S100A8和S100A9表达的影响
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摘要
目的金黄色葡萄球菌肠毒素B(Staphylococcus aureus enterotoxin B,SEB)刺激HaCaT细胞后观察S100A8、S100A9、白介素-1α(IL-1ɑ)和白介素-17 (IL-17)表达的影响。进一步阐明机体组织损伤相关分子模式(damage associated molecularpatterns,DAMP)与先天免疫对AD发病的影响。方法采用MTT法检测SEB对HaCaT细胞的细胞毒性。通过蛋白免疫印迹法测定细胞内外DAMP分子S100A8、S100A9蛋白表达。用酶联免疫吸附实验检测HaCaT细胞培养液中的IL-1α、IL-17的表达情况。结果在24、48、72 h,不同浓度SEB对HaCaT细胞无毒性作用。SEB刺激组细胞内外S100A8、S100A9蛋白表达较对照组增多。在不同浓度的SEB处理下IL-1α表达水平呈上升趋势。100 ng/mL SEB刺激组较对照组相比,IL-17表达水平明显升高。结论 SEB可能通过上调DAMP分子S100A8、S100A9蛋白以及IL-1α、IL-17炎症因子的表达,参与AD的发病机制。
Objective To detect the S100A8,S100A9,IL-1 a and IL-17 expression in the Staph-ylococcus aureus enterotoxin B( SEB) stimulated HaCaT cells,and clarify the impactof damage associated molecular patterns( DAMPs) and innate immunity on the patho-genesis of AD. Methods MTT assay was used to determine the cytotoxicity of SEBon the HaCaT cells. The expression of DAMP molecules S100A8 and S100A9 pro-teins were detected by Western blot. The expression of IL-1α and IL-17 were meas-ured by enzyme linked immunosorbent assay( ELISA). Results Different concen-trations of SEB had no cytotoxicity on Ha Ca T cells at 24,48 and 72 h. The expressionof intracellular and extracellular S100A8 and S100A9 were significantly increased in theSEB-treat group. The expression of IL-1α was increased in the SEB-treat Ha Ca T cellsin concentration dependent manner. The expression of IL-17 was significantly increasedin the 100 ng/mL SEB-treat group. Conclusion SEB participates the pathogenesis ofAD through upregulating S100A8,S100A9,IL-1α and IL-17.
Objective To detect the S100A8,S100A9,IL-1 a and IL-17 expression in the Staph-ylococcus aureus enterotoxin B( SEB) stimulated HaCaT cells,and clarify the impactof damage associated molecular patterns( DAMPs) and innate immunity on the patho-genesis of AD. Methods MTT assay was used to determine the cytotoxicity of SEBon the HaCaT cells. The expression of DAMP molecules S100A8 and S100A9 pro-teins were detected by Western blot. The expression of IL-1α and IL-17 were meas-ured by enzyme linked immunosorbent assay( ELISA). Results Different concen-trations of SEB had no cytotoxicity on Ha Ca T cells at 24,48 and 72 h. The expressionof intracellular and extracellular S100A8 and S100A9 were significantly increased in theSEB-treat group. The expression of IL-1α was increased in the SEB-treat Ha Ca T cellsin concentration dependent manner. The expression of IL-17 was significantly increasedin the 100 ng/mL SEB-treat group. Conclusion SEB participates the pathogenesis ofAD through upregulating S100A8,S100A9,IL-1α and IL-17.
引文
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[2]Bieber T.Atopic dermatitis 2.0:from the clinical phenotype to the molecular taxonomy and stratified medicine[J].Allergy,2012,67(12):1475-1482.
[3]Gomes PL,Malaviqe GN,Fernando N,et al.Characteristics of staphylococcus aureus colonization in patients with atopic dermatitis in Sri Lanka[J].Clini Exp Dermatol,2011,36(2):195-200.
[4]Park HY,Kim CR,Huh IS,et al.Straphylococcus aureus colonization in acute and chronic skin lesions of patients with atopic dermatitis[J].Ann Dermatol,2013,25(4):410-416.
[5]Totte JE,van der Feltz WT,Hennekam M,et al.Prevalence and odds of Staphylococcus aureus carriage in atopic dermatitis:A systematic review and meta-analysis[J].Br JDermatol,2016,175(4):687-695.
[6]Hong SW,Choi EB,Min TK,et al.An important role ofα-hemolysin in extracellular vesicles on the development of atopic dermatitis induced by Staphylococcus aureus[J].PLo S one,2014,9(7):epub.
[7]Morizane S,Gallo RL.Antimicrobial peptides in the pathogenesis of psoriasis[J].J Invest Dermatol,2012,39(3):225-230.
[8]Michelle MM,Kerkhoff C,Bornfeldt KE.S100A8 and S100A9 in cardiovascular biology and disease[J].Arterioscler Thromb Vasc Biol,2012,32(2):223-229.
[9]D'Amico F,Granata M,Skarmoutsou E,et al.Biological therapy downregulates the heterodimer S100A8/A9(calprotectin)expression in psoriatic patients[J].Inflamm Res,2018,67(7):609-616.
[10]Suarez-Farinas M,Dhingra N,Gittler J,et al.Intrinsic atopic dermatitis shows similar TH2 and higher TH17 immune activation compared with extrinsic atopic dermatitis[J].J Allergy Clin Immunol,2013,132(2):361-370.
[11]Jin S,Park CO,Shin JU,et al.DAMP molecules S100A9and S100A8 activated by IL-17A and house-dust mites are increased in atopic dermatitis[J].Exp Dermatol,2014,23(12):938-941.
[12]Mose M,Kang Z,Raaby L,et al.TNFα-and IL-17A-mediated S100A8 expression is regulated by p38 MAPK[J].Exp Dermatol,2013,22(7):476-481.
[13]He R,Oyoshi MK,Jin H,et al.Epicutaneous antigen exposure induces a Th17 response that drives airway inflammation after inhalation challenge[J].Proc Natl Acad Sci USA,2007,104(40):15817-15822.
[14]Kim KH,Han JH,Chung JH,et al.Role of staphylococcal superantigen in atopic dermatitis:influence on keratinocytes[J].J Korean Med Sci,2006,21(2):315-323.
[15]Nakatsuji T,Chen TH,Two AM,et al.Staphylococcus aureus exploits epidermal barrier defects in atopic dermatitis to trigger cytokine expression[J].J Invest Dermatol,2016,136(11):2192-2200.