中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2009年
6期
409-412
,共4页
陆小年%张勇%韩凌%祝禄川%郑志忠
陸小年%張勇%韓凌%祝祿川%鄭誌忠
륙소년%장용%한릉%축록천%정지충
ToIl样受体2%角蛋白细胞%细胞增殖%NF-KB%转化生长因子α
ToIl樣受體2%角蛋白細胞%細胞增殖%NF-KB%轉化生長因子α
ToIl양수체2%각단백세포%세포증식%NF-KB%전화생장인자α
Toll-like receptor 2%Keratinocytes%Cell proliferation%NF-kappa B:Transforming growth factor alpha
目的 探讨Toll样受体2(TLR2)对体外培养人角质形成细胞增殖的影响.方法 天然配体肽聚糖(PGN)体外活化人角质形成细胞TLR2,用噻唑蓝法检测PGN对角质形成细胞体外增殖的影响,确定最适作用浓度;用实时荧光定量PCR法和Western印迹法分别检测角质形成细胞Ki67、TLR2、核因子(NF)-KBp65及转化生长因子(TGF)-α mRNA和蛋白的表达水平;采用抗体封闭实验分析封闭TLR2对PGN诱导角质形成细胞Ki67、TLR2、NF-KBp65及TGF-α表达的影响.结果 不同浓度的PGN处理角质形成细胞后24 h,在1.25,2.5,5 μg/mL的浓度下细胞较对照组出现明显增殖(P＜0.05). 1.25、2.5和5μg/mL PGN作用于角质形成细胞后24 h,其中1.25和2.5 μg/mL浓度下Ki67 mRNA的表达明显增加,各组Ki67蛋白合成均有增加;各组TLR2 mRNA和蛋白的表达均增加;1.25μg/mL浓度下NF-KBp65 mRNA的表达明显增加,各组细胞核内NF-KBp65蛋白合成均增加;其中1.25和5 μg/mL浓度下TGF-α mRNA和蛋白表达均明显增加(P＜0.05).以抗人TLR2单克隆阻断性抗体封闭TLR2后,PGN诱导的角质形成细胞Ki67、TI,R2、NF-KBp65及TGF-α mRNA和蛋白表达的上调均受到明显抑制(P＜0.05).结论 角质形成细胞TLR2经PGN诱导活化后,可能通过促进NF-KB活化及TGF-α表达而导致角质形成细胞的异常增殖.
目的 探討Toll樣受體2(TLR2)對體外培養人角質形成細胞增殖的影響.方法 天然配體肽聚糖(PGN)體外活化人角質形成細胞TLR2,用噻唑藍法檢測PGN對角質形成細胞體外增殖的影響,確定最適作用濃度;用實時熒光定量PCR法和Western印跡法分彆檢測角質形成細胞Ki67、TLR2、覈因子(NF)-KBp65及轉化生長因子(TGF)-α mRNA和蛋白的錶達水平;採用抗體封閉實驗分析封閉TLR2對PGN誘導角質形成細胞Ki67、TLR2、NF-KBp65及TGF-α錶達的影響.結果 不同濃度的PGN處理角質形成細胞後24 h,在1.25,2.5,5 μg/mL的濃度下細胞較對照組齣現明顯增殖(P＜0.05). 1.25、2.5和5μg/mL PGN作用于角質形成細胞後24 h,其中1.25和2.5 μg/mL濃度下Ki67 mRNA的錶達明顯增加,各組Ki67蛋白閤成均有增加;各組TLR2 mRNA和蛋白的錶達均增加;1.25μg/mL濃度下NF-KBp65 mRNA的錶達明顯增加,各組細胞覈內NF-KBp65蛋白閤成均增加;其中1.25和5 μg/mL濃度下TGF-α mRNA和蛋白錶達均明顯增加(P＜0.05).以抗人TLR2單剋隆阻斷性抗體封閉TLR2後,PGN誘導的角質形成細胞Ki67、TI,R2、NF-KBp65及TGF-α mRNA和蛋白錶達的上調均受到明顯抑製(P＜0.05).結論 角質形成細胞TLR2經PGN誘導活化後,可能通過促進NF-KB活化及TGF-α錶達而導緻角質形成細胞的異常增殖.
목적 탐토Toll양수체2(TLR2)대체외배양인각질형성세포증식적영향.방법 천연배체태취당(PGN)체외활화인각질형성세포TLR2,용새서람법검측PGN대각질형성세포체외증식적영향,학정최괄작용농도;용실시형광정량PCR법화Western인적법분별검측각질형성세포Ki67、TLR2、핵인자(NF)-KBp65급전화생장인자(TGF)-α mRNA화단백적표체수평;채용항체봉폐실험분석봉폐TLR2대PGN유도각질형성세포Ki67、TLR2、NF-KBp65급TGF-α표체적영향.결과 불동농도적PGN처리각질형성세포후24 h,재1.25,2.5,5 μg/mL적농도하세포교대조조출현명현증식(P＜0.05). 1.25、2.5화5μg/mL PGN작용우각질형성세포후24 h,기중1.25화2.5 μg/mL농도하Ki67 mRNA적표체명현증가,각조Ki67단백합성균유증가;각조TLR2 mRNA화단백적표체균증가;1.25μg/mL농도하NF-KBp65 mRNA적표체명현증가,각조세포핵내NF-KBp65단백합성균증가;기중1.25화5 μg/mL농도하TGF-α mRNA화단백표체균명현증가(P＜0.05).이항인TLR2단극륭조단성항체봉폐TLR2후,PGN유도적각질형성세포Ki67、TI,R2、NF-KBp65급TGF-α mRNA화단백표체적상조균수도명현억제(P＜0.05).결론 각질형성세포TLR2경PGN유도활화후,가능통과촉진NF-KB활화급TGF-α표체이도치각질형성세포적이상증식.
Objective To investigate the effect of Toll-like receptor 2(TLR2)on the proliferation of human keratinocytes.Methods Keratinocytes were isolated from the foreskin of children,and subjected to primary culture.Atier 3-5 passages.the kemtinocytes were incubated with various concentrations of peptidoglycan(PGN).a TLR2 agonist.Cell proliferation was detected by MTT colorimetric assay and the suitable concentrations of PGN were determined.The mRNA and protein expressions of Ki67.TLR2.NF-kB p65 and TGF-α were detected by real-time quantitative PCR and Western blot.respectively,in keratinocytes treated witll PGN of 0,1.25,2.5 and 5 μg/mL.Antibody blocking test was utilized to evaluate the effect of blocking TLR2 with specific anti-TLR2 neutralizing monoclonal antibody before incubation with PGN on the expressions of Ki67,TLR2,NF-KB p65 and TGF-α by keratinocytes.Results The proliferation of kemtinocytes was significantly promoted by the incubation with PGN of 1.25,2.5 and 5μg/mL for 24 hours (all P＜0.05),which also increased the expression of Ki67 protein,TLR2 mRNA and protein,and NF-KB p65 protein.Further more,the mRNA expression of Ki67 in keratinocytes was elevated bv PGN of 1.25 and 2.5μg/mL,the mRNA expression of NF-KB p65 elevated by PGN of 1.25μg/mL,and the expressions of TGF-αprotein and mRNA elevated by PGN of 1.25 and 5μg/mL (P＜0.05).The mRNA and protein expressions of Ki67,TLR2,NF-kB p65 and TGF-αwere all inhibited by the blocking of TLR2 before incubation with PGN (a11 P＜0.05).Conclusion Activation of TLR2 bv PGN could induce the over-proliferation of human keratinocytes,likely through promoting NF-rB activation and TGF-α expression.
