中文摘要：

目的探讨炎症作用下NF-κB通路对人牙龈间充质干细胞（GMSCs）分泌炎性因子能力的影响。方法体外分离纯化培养正常组织来源的GMSCs。在相同培养条件下,GMSCs被分成3组。应用梯度浓度TNF-α（0、10、20 ng/ml）诱导12 h。Annexin V-FITC/PI双染检测3组间GMSCs凋亡率差异。应用Western blot检测NLK、I-κBα、P-IκBα在NF-κB激活通路中的表达,Real-Time PCR检测TNF-α、IL-1β在GMSCs加入抑制剂BAY 11-7082后基因表达变化。结果凋亡检测显示,GMSCs凋亡率与TNF-α浓度呈正相关（P〈0.05）;Western blot结果显示,与对照组相比,NLK、I-κBα表达随TNF-α浓度增加而降低（P〈0.05）,而P-IκBα表达量呈显著增高趋势（P〈0.01）。PCR检测结果显示,随外源性TNF-α浓度升高,10 ng/ml组与20 ng/ml组GMSCs的TNF-α与IL-1β基因表达量也显著升高（P〈0.01）。TNF-α诱导组加入NF-κB抑制剂BAY 11-7082后检测GMSCs,10 ng/ml组与20 ng/ml组IL-1β与TNF-α基因表达量下降,与对照组比较差异无统计学意义（P〉0.05）。结论 GMSCs中NLK在炎性环境下表达量降低,炎性因子可以通过NF-κB通路增强牙龈间充质干细胞中TNF-α和IL-1β的表达。

英文摘要：

Objective To investigate the effects of NF-kappa B pathway on the secretion ability of inflammatory cytokines in human gingival mesenchymal stem cells（GMSCs）. Methods GMSCs derived from normal tissues were isolated and purified in vitro.GMSCs were divided into three groups under the same culture conditions and induced by TNF-α（0,10 ng/ml,and 20 ng/ml）for 12 h.The difference of apoptosis rate between the three groups was detected by Annexin V-FITC/PI double staining. Western blot analysis was used to detect the expression of NLK, I-κBα, and P-IκBα in the activation pathway of NF-κB. The gene expression changes of TNF-α and IL-1β in GMSCs after adding the inhibitor of BA Y-11-7082 were detected by Real-Time PCR. Results Apoptosis detection showed that the apoptosis rate of GMSCs was positively correlated with the concentration of TNF-α. Western blot results showed that compared with the expression in the control group, the expression of NLK and I-κBα decreased along with the increase of TNF-α concentration（P 0.05）. P-IκBα expression had the obvious tendency of increase（P 0.01）. The PCR results indicated that along with the increase of exogenous concentration, the TNF-α and IL-1β gene expression significantly increased in 10 ng/ml group and 20 ng/ml group（P 0.01）. After the adding of inhibitor BAY 11-7082, the expression of IL-1β and TNF-α decreased in 10 ng/ml group and 20 ng/ml group, but the differences between those groups and the control group were not significant（P 0.05）.Conclusion NLK expression decreases in inflammatory environment. The inflammatory factors can enhance the expression of TNF-αand IL-1β in gingival mesenchymal stem cells through the NF-κB pathway.