全科口腔医学电子杂志
全科口腔醫學電子雜誌
전과구강의학전자잡지
2014年
1期
13-17,41
,共6页
杜星颜%陈莉莉%陈洁%刘加荣
杜星顏%陳莉莉%陳潔%劉加榮
두성안%진리리%진길%류가영
人牙龈上皮细胞%IL-1β%TNF-α%β-防御素%信号通路
人牙齦上皮細胞%IL-1β%TNF-α%β-防禦素%信號通路
인아간상피세포%IL-1β%TNF-α%β-방어소%신호통로
Human gingival epithelial cells%IL-1β%TNF-α%β-defensins%Signaling pathways
目的：研究牙龈上皮细胞在炎症因子IL-1β，TNF-α诱导下分泌β-防御素1，2，3的信号通路。方法取30岁以下因拔除第三磨牙或助萌需切除的牙龈，原代培养人牙龈上皮细胞，并用10μmol/L MAPK抑制剂SB203580或NF-κB抑制剂BAY 11-7082孵育2 h，而后用150 ng/ml IL-1β或TNF-α诱导抑制剂孵育过的细胞24 h，实时荧光定量PCR检测β-防御素1，2，3的基因表达水平。结果在MAPK或NF-κB抑制剂存在的情况下，人牙龈上皮细胞经炎症因子IL-1β，TNF-α诱导后，HBD-1的表达量无明显变化；但HBD-2的表达量明显降低，MAPK抑制剂使牙龈上皮细胞在IL-1β和TNF-α诱导下分泌HBD-2的量分别下降81%和76%，NF-κB抑制剂使牙龈上皮细胞在IL-1β和TNF-α诱导下分泌HBD-2的量分别下降93%和95%；两种抑制剂对HBD-3的表达量表现出不同效应， MAPK抑制剂使牙龈上皮细胞在IL-1β和TNF-α诱导下分泌HBD-3的量分别下降65%和66%，而NF-κB抑制剂对HBD-3的表达量无明显影响。结论人牙龈上皮细胞较稳定的分泌HBD-1，不受外界炎症因子及信号通路抑制剂的影响。MAPK和NF-κB信号通路在HBD-2的诱导分泌中起了非常重要的作用；本研究中MAPK信号通路与HBD-3的诱导表达密切相关。
目的：研究牙齦上皮細胞在炎癥因子IL-1β，TNF-α誘導下分泌β-防禦素1，2，3的信號通路。方法取30歲以下因拔除第三磨牙或助萌需切除的牙齦，原代培養人牙齦上皮細胞，併用10μmol/L MAPK抑製劑SB203580或NF-κB抑製劑BAY 11-7082孵育2 h，而後用150 ng/ml IL-1β或TNF-α誘導抑製劑孵育過的細胞24 h，實時熒光定量PCR檢測β-防禦素1，2，3的基因錶達水平。結果在MAPK或NF-κB抑製劑存在的情況下，人牙齦上皮細胞經炎癥因子IL-1β，TNF-α誘導後，HBD-1的錶達量無明顯變化；但HBD-2的錶達量明顯降低，MAPK抑製劑使牙齦上皮細胞在IL-1β和TNF-α誘導下分泌HBD-2的量分彆下降81%和76%，NF-κB抑製劑使牙齦上皮細胞在IL-1β和TNF-α誘導下分泌HBD-2的量分彆下降93%和95%；兩種抑製劑對HBD-3的錶達量錶現齣不同效應， MAPK抑製劑使牙齦上皮細胞在IL-1β和TNF-α誘導下分泌HBD-3的量分彆下降65%和66%，而NF-κB抑製劑對HBD-3的錶達量無明顯影響。結論人牙齦上皮細胞較穩定的分泌HBD-1，不受外界炎癥因子及信號通路抑製劑的影響。MAPK和NF-κB信號通路在HBD-2的誘導分泌中起瞭非常重要的作用；本研究中MAPK信號通路與HBD-3的誘導錶達密切相關。
목적：연구아간상피세포재염증인자IL-1β，TNF-α유도하분비β-방어소1，2，3적신호통로。방법취30세이하인발제제삼마아혹조맹수절제적아간，원대배양인아간상피세포，병용10μmol/L MAPK억제제SB203580혹NF-κB억제제BAY 11-7082부육2 h，이후용150 ng/ml IL-1β혹TNF-α유도억제제부육과적세포24 h，실시형광정량PCR검측β-방어소1，2，3적기인표체수평。결과재MAPK혹NF-κB억제제존재적정황하，인아간상피세포경염증인자IL-1β，TNF-α유도후，HBD-1적표체량무명현변화；단HBD-2적표체량명현강저，MAPK억제제사아간상피세포재IL-1β화TNF-α유도하분비HBD-2적량분별하강81%화76%，NF-κB억제제사아간상피세포재IL-1β화TNF-α유도하분비HBD-2적량분별하강93%화95%；량충억제제대HBD-3적표체량표현출불동효응， MAPK억제제사아간상피세포재IL-1β화TNF-α유도하분비HBD-3적량분별하강65%화66%，이NF-κB억제제대HBD-3적표체량무명현영향。결론인아간상피세포교은정적분비HBD-1，불수외계염증인자급신호통로억제제적영향。MAPK화NF-κB신호통로재HBD-2적유도분비중기료비상중요적작용；본연구중MAPK신호통로여HBD-3적유도표체밀절상관。
Objective The purpose of this study is to investigate the signaling pathways involved in humanβ-defensins (HBD-1,2,3) induction in response to IL-1βand TNF-αin human gingival epithelial cells. Methods Healthy gingival samples were obtained from patients less than 30 years old undergoing the third-molar extraction or impacted teeth exposure. Primary human epithelial cells were cultured, then the cells were pretreated with 10μmol/L MAPK or NF-κB inhibitor for 2 hours and cultured further with 150 ng/ml IL-1β or TNF-α. Quantitative real-time PCR was utilized to quantify the HBD-1,2,3 mRNA expression. Results HBD-1 induction by IL-1βwas not blocked in the presence of MAPK or NF-κB inhibitor. The HBD-1 induction by TNF-αwas similar to that of IL-1β. In contrast, there was a notable decrease in HBD-2 induction by IL-1βand the reduction was 81%by MAPK inhibitor and 93%by NF-κB inhibitor. Similarly, the HBD-2 induction by TNF-α was inhibited by 76% by MAPK inhibitor and 95% by NF-κB inhibitor. HBD-3 induction was different from HBD-2. The HBD-3 expression level induced by IL-1βwas decreased by 65%in the presence of MAPK inhibitor and induced expression by TNF-α was decreased 66%. Whereas the NF-κB inhibitor showed no inhibition on HBD-3 induction both by IL-1β and TNF-α. Conclusion Our results suggested that different signaling pathways were involved in the induction of humanβ-defensins in human gingival epithelial cells stimulated by IL-1βand TNF-α. For induced HBD-1 expression by IL-1βand TNF-α, neither MAPK nor NF-κB was essential or sufficient. Both MAPK and NF-κB signaling pathways played an important role in the HBD-2 induction by IL-1βand TNF-α. Whereas the NF-κB inhibitor showed no inhibition on HBD-3 induction both by IL-1βand TNF-α.
