Objective Proprionibacterium acnes(P.acnes)-induced inflammatory responses,proliferation and differentiation of keratinocytes contribute to the progression of acne vulgaris(AV).P.acnes was found to enhance the product...Objective Proprionibacterium acnes(P.acnes)-induced inflammatory responses,proliferation and differentiation of keratinocytes contribute to the progression of acne vulgaris(AV).P.acnes was found to enhance the production of interleukin-8(IL-8)by keratinocytes.This study aimed to investigate the role of IL-8 in P.acnes-induced proliferation and differentiation of keratinocytes and the underlying mechanism.Methods The P.acnes-stimulated HaCaT cell(a human keratinocyte cell line)model was established.Western blotting and immunofluorescence were performed to detect the expression of the IL-8 receptors C-X-C motif chemokine receptor 1(CXCR1)and C-X-C motif chemokine receptor 2(CXCR2)on HaCaT cells.Cell counting kit-8(CCK-8)assay,5-ethynyl-20-deoxyuridine(EdU)assay and Western blotting were performed to examine the effects of IL-8/CXCR2 axis on the proliferation and differentiation of HaCaT cells treated with P.acnes,the IL-8 neutralizing antibody,the CXCR2 antagonist(SB225002),or the CXCR1/CXCR2 antagonist(G31P).Western blotting,nuclear and cytoplasmic separation,CCK-8 assay,and EdU assay were employed to determine the downstream pathway of CXCR2 after P.acnes-stimulated HaCaT cells were treated with the CXCR2 antagonist,the protein kinase B(AKT)antagonist(AZD5363),or the constitutively active forkhead box O1(FOXO1)mutant.Finally,autophagy markers were measured in HaCaT cells following the transfection of the FOXO1 mutant or treatment with the autophagy inhibitor 3-methyladenine(3-MA).Results The expression levels of CXCR1 and CXCR2 were significantly increased on the membrane of HaCaT cells following P.acnes stimulation.The IL-8/CXCR2 axis predominantly promoted the proliferation and differentiation of P.acnes-induced HaCaT cells by activating AKT/FOXO1/autophagy signaling.In brief,IL-8 bound to its receptor CXCR2 on the membrane of keratinocytes to activate the AKT/FOXO1 axis.Subsequently,phosphorylated FOXO1 facilitated autophagy to promote the proliferation and differentiation of P.acnes-induced keratinocytes.Conclusion This study demonstrated the novel autocrine effect of IL-8 on the proliferation and differentiation of P.acnes-induced keratinocytes,suggesting a potential therapeutic target for AV.展开更多
AIM:To detect the quantitative expression levels of the pro-inflammatory interleukin-8(IL8),antimicrobial peptides human beta defense-2(HBD2),and human beta defense-3(HBD3)genes in bacterial conjunctivitis.METHODS:The...AIM:To detect the quantitative expression levels of the pro-inflammatory interleukin-8(IL8),antimicrobial peptides human beta defense-2(HBD2),and human beta defense-3(HBD3)genes in bacterial conjunctivitis.METHODS:The human conjunctival epithelial cells were obtained using the impression cytology technique from healthy controls and patients.The genes expression levels were determined utilizing a reverse transcription quantitative polymerase chain reaction(RT-q PCR).The contribution of causative agent type,the number of isolates and severity of clinical features,in the increase of genes expression was also determined.RESULTS:The RT-q PCR showed that IL8,HBD2,and HBD3 expression increased in bacterial conjunctivitis as compared to healthy control(P<0.001).In gram-negative bacterial conjunctivitis,HBD2 was highly up-regulated(P<0.001)compared to other types of bacterial conjunctivitis.In mixed bacterial conjunctivitis,a direct correlation between HBD2 up-regulation and HBD3 up-regulation was observed(P<0.05).The severity of clinical features was related to the up-regulation of IL8 and HBD2(P<0.05).CONCLUSION:IL8,HBD2,and HBD3 are immuneeffectors in infectious conjunctivitis.HBD2 is active during different bacterial conjunctivitis but is more released with gram-negative bacteria compared to gram-positive bacteria.HBD3 is an obvious defender in different bacterial conjunctivitis.展开更多
基金supported by the National Natural Science Foundation of China(No.82103756).
文摘Objective Proprionibacterium acnes(P.acnes)-induced inflammatory responses,proliferation and differentiation of keratinocytes contribute to the progression of acne vulgaris(AV).P.acnes was found to enhance the production of interleukin-8(IL-8)by keratinocytes.This study aimed to investigate the role of IL-8 in P.acnes-induced proliferation and differentiation of keratinocytes and the underlying mechanism.Methods The P.acnes-stimulated HaCaT cell(a human keratinocyte cell line)model was established.Western blotting and immunofluorescence were performed to detect the expression of the IL-8 receptors C-X-C motif chemokine receptor 1(CXCR1)and C-X-C motif chemokine receptor 2(CXCR2)on HaCaT cells.Cell counting kit-8(CCK-8)assay,5-ethynyl-20-deoxyuridine(EdU)assay and Western blotting were performed to examine the effects of IL-8/CXCR2 axis on the proliferation and differentiation of HaCaT cells treated with P.acnes,the IL-8 neutralizing antibody,the CXCR2 antagonist(SB225002),or the CXCR1/CXCR2 antagonist(G31P).Western blotting,nuclear and cytoplasmic separation,CCK-8 assay,and EdU assay were employed to determine the downstream pathway of CXCR2 after P.acnes-stimulated HaCaT cells were treated with the CXCR2 antagonist,the protein kinase B(AKT)antagonist(AZD5363),or the constitutively active forkhead box O1(FOXO1)mutant.Finally,autophagy markers were measured in HaCaT cells following the transfection of the FOXO1 mutant or treatment with the autophagy inhibitor 3-methyladenine(3-MA).Results The expression levels of CXCR1 and CXCR2 were significantly increased on the membrane of HaCaT cells following P.acnes stimulation.The IL-8/CXCR2 axis predominantly promoted the proliferation and differentiation of P.acnes-induced HaCaT cells by activating AKT/FOXO1/autophagy signaling.In brief,IL-8 bound to its receptor CXCR2 on the membrane of keratinocytes to activate the AKT/FOXO1 axis.Subsequently,phosphorylated FOXO1 facilitated autophagy to promote the proliferation and differentiation of P.acnes-induced keratinocytes.Conclusion This study demonstrated the novel autocrine effect of IL-8 on the proliferation and differentiation of P.acnes-induced keratinocytes,suggesting a potential therapeutic target for AV.
文摘AIM:To detect the quantitative expression levels of the pro-inflammatory interleukin-8(IL8),antimicrobial peptides human beta defense-2(HBD2),and human beta defense-3(HBD3)genes in bacterial conjunctivitis.METHODS:The human conjunctival epithelial cells were obtained using the impression cytology technique from healthy controls and patients.The genes expression levels were determined utilizing a reverse transcription quantitative polymerase chain reaction(RT-q PCR).The contribution of causative agent type,the number of isolates and severity of clinical features,in the increase of genes expression was also determined.RESULTS:The RT-q PCR showed that IL8,HBD2,and HBD3 expression increased in bacterial conjunctivitis as compared to healthy control(P<0.001).In gram-negative bacterial conjunctivitis,HBD2 was highly up-regulated(P<0.001)compared to other types of bacterial conjunctivitis.In mixed bacterial conjunctivitis,a direct correlation between HBD2 up-regulation and HBD3 up-regulation was observed(P<0.05).The severity of clinical features was related to the up-regulation of IL8 and HBD2(P<0.05).CONCLUSION:IL8,HBD2,and HBD3 are immuneeffectors in infectious conjunctivitis.HBD2 is active during different bacterial conjunctivitis but is more released with gram-negative bacteria compared to gram-positive bacteria.HBD3 is an obvious defender in different bacterial conjunctivitis.
