中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2011年
4期
289-293
,共5页
刘佳明%屠燕烨%郭雅君%丁卉%楼永良
劉佳明%屠燕燁%郭雅君%丁卉%樓永良
류가명%도연엽%곽아군%정훼%루영량
乳杆菌细胞壁成分%人阴道上皮细胞%人β-防御素2%p38MAPK%NF-κB
乳桿菌細胞壁成分%人陰道上皮細胞%人β-防禦素2%p38MAPK%NF-κB
유간균세포벽성분%인음도상피세포%인β-방어소2%p38MAPK%NF-κB
Lactobacillus cell wall extract%Human vaginal epithelial cell%Human-β-defensin-2%p38 MAPK%NF-κB
目的 探讨乳杆菌细胞肇成分(Lactobacillus cell wall extract,LCWE)对人阴道上皮细胞β-防御素-2的诱导作用及细胞内信号转导机制.方法 采用实时定量PCR和Western blot方法检测LCWE处理对人阴道上皮细胞(WZV-1)β-防御素-2表达的影响;Western blot方法检测LCWE处理后WZV-1细胞内NF-κB和p38MAPK信号通路活化情况;实时定量PCR和Western blot方法检测NF-κB抑制剂和p38MAPK抑制剂预处理WZV-1细胞对LCWE诱导β-防御素-2表达的影响.结果 LCWE可诱导WZV-1细胞β-防御素-2表达且存在着明显的时间效应和剂量依赖关系,50μg/mlLCWE处理细胞8 h对β-防御素-2的上调幅度最大,刺激0.5 h后可引起细胞内p38MAPK蛋白的磷酸化显著增加,1 h达到顶峰;刺激0.5 h后可引起细胞核内NF-κB含量显著增加(P<0.05),2 h达到顶峰.NF-κB抑制剂PDTC、p38MAPK抑制剂SB20380预处理WZV-1细胞,可明显抑制LCWE诱导WZV-1细胞β-防御素-2的表达.结论 LCWE具有激活NF-κB和p38MAPK信号通路诱导阴道-上皮细胞β-防御素-2的作用.
目的 探討乳桿菌細胞肇成分(Lactobacillus cell wall extract,LCWE)對人陰道上皮細胞β-防禦素-2的誘導作用及細胞內信號轉導機製.方法 採用實時定量PCR和Western blot方法檢測LCWE處理對人陰道上皮細胞(WZV-1)β-防禦素-2錶達的影響;Western blot方法檢測LCWE處理後WZV-1細胞內NF-κB和p38MAPK信號通路活化情況;實時定量PCR和Western blot方法檢測NF-κB抑製劑和p38MAPK抑製劑預處理WZV-1細胞對LCWE誘導β-防禦素-2錶達的影響.結果 LCWE可誘導WZV-1細胞β-防禦素-2錶達且存在著明顯的時間效應和劑量依賴關繫,50μg/mlLCWE處理細胞8 h對β-防禦素-2的上調幅度最大,刺激0.5 h後可引起細胞內p38MAPK蛋白的燐痠化顯著增加,1 h達到頂峰;刺激0.5 h後可引起細胞覈內NF-κB含量顯著增加(P<0.05),2 h達到頂峰.NF-κB抑製劑PDTC、p38MAPK抑製劑SB20380預處理WZV-1細胞,可明顯抑製LCWE誘導WZV-1細胞β-防禦素-2的錶達.結論 LCWE具有激活NF-κB和p38MAPK信號通路誘導陰道-上皮細胞β-防禦素-2的作用.
목적 탐토유간균세포조성분(Lactobacillus cell wall extract,LCWE)대인음도상피세포β-방어소-2적유도작용급세포내신호전도궤제.방법 채용실시정량PCR화Western blot방법검측LCWE처리대인음도상피세포(WZV-1)β-방어소-2표체적영향;Western blot방법검측LCWE처리후WZV-1세포내NF-κB화p38MAPK신호통로활화정황;실시정량PCR화Western blot방법검측NF-κB억제제화p38MAPK억제제예처리WZV-1세포대LCWE유도β-방어소-2표체적영향.결과 LCWE가유도WZV-1세포β-방어소-2표체차존재착명현적시간효응화제량의뢰관계,50μg/mlLCWE처리세포8 h대β-방어소-2적상조폭도최대,자격0.5 h후가인기세포내p38MAPK단백적린산화현저증가,1 h체도정봉;자격0.5 h후가인기세포핵내NF-κB함량현저증가(P<0.05),2 h체도정봉.NF-κB억제제PDTC、p38MAPK억제제SB20380예처리WZV-1세포,가명현억제LCWE유도WZV-1세포β-방어소-2적표체.결론 LCWE구유격활NF-κB화p38MAPK신호통로유도음도-상피세포β-방어소-2적작용.
Objective To investigate the molecular and cell signal transduction mechanisms by Lactobacillus cell wall extract(LCWE)inducing human-β-defensin-2(hBD-2)expression in human vaginal epithelial cells.Methods The induction of hBD-2 in human vaginal epithelial cells(WZV-1)by LCWE was observed using real-time PCR and Western blot.After stimulating WZV-1.the activation of NF-κB and p38MAPK signaling pathways were determined by Western blot.The induction of hBD-2 in WZV-1 cells by LCWE was observed with signaling pathways inhibitors of NF-κB and p38MAPK using real-time PCR and Western blot.Results The results showed that LCWE significantly upregulated hBD-2 expression in the time and dose-dependent manner.The maximal stimulatory effect of LCWE on the expression of hBD-2mRNA in WZV-1 cells were observed at the concentration of 50μg/ml after treatment for 8 h.After stimulation by 50μg/ml LCWE,Western blot analysis demonstrated that the phosphorylation of p38MAPK increased at 0.5 h significantly,peaked at 1 h,moreover the concentration of NF-κB in nucleus increased at 0.5 h significantly(P<0.05),peaked at 2 h.Blocking with inhibitor of NF-κB and(or)p38MAPK pathways results in decreased levels of HBD-2 expression.Conclusion These findings suggest that p38MAPK and NF-κB pathways play the important roles in induction of hBD-2 expression by LCWE in human vagihal epithelial cells.
