摘要
目的探讨JAK2/STAT3信号通路是否影响低氧诱导的人单核细胞THP-1的炎症因子表达。方法体外低氧(1%O2)培养THP-1细胞24 h构建细胞氧化损伤的实验模型。ELISA法检测IL-1β和TNF-α的分泌水平,Westernblot检测p-JAK2、p-STAT3、JAK2、STAT3和细胞核内NF-κB p65的蛋白表达,实时荧光定量PCR检测炎症因子IL-1β、TNF-α的m RNA表达水平。结果与空白对照组相比,低氧诱导THP-1细胞24 h后,IL-1β、TNF-α的分泌和m RNA表达水平增加,差异均有统计学意义(P<0.05);细胞内p-JAK2、p-STAT3和胞核NF-κB p65蛋白表达增加,差异具有统计学意义(P<0.05)。使用JAK2抑制剂AG490能明显抑制低氧诱导的THP-1细胞的p-JAK2、p-STAT3和胞核NF-κBp65的蛋白表达,以及能够降低IL-1β、TNF-α的分泌和m RNA表达水平,差异均具有统计学意义(P<0.05)。结论低氧可诱导细胞炎症因子的表达增加,其机制可能与激活JAK2/STAT3信号通路有关。
Objective To investigate whether the JAK2/STAT3 signaling pathway affects the expression of inflammatorycytokines in human monocytes THP-1 induced by hypoxia. Methods THP-1 cells were cultured under hypoxia（1% O2） for24 h to build a cellular oxidative damage model. The concentrations of interleutin-1 beta（IL-1β） and tumor necrosis factor-α（TNF-α） in the supernatant were determined by ELISA. The protein levels of p-JAK2,p-STAT3,JAK2,STAT3 and nuclearNF-κB p65 expression were detected by Western blotting. The m RNA expression of IL-1β and TNF-α was evaluated by q RT-PCR. Results Compared with the blank control cells,the inducement of hypoxic condition for 24 h resulted in the significantincreases of the secretion and the m RNA levels of IL- 1β and TNF-α（P〈0.05）,and obvious elevations in the intracellularexpression of p-JAK2 and p-STAT3 and in the nuclear level of NF-κB p65 in THP-1 cells（P〈0.05）. However,JAK2 inhibitorAG490 could significantly inhibit the protein levels of p-JAK2,p-STAT3 and nuclear NF- κB p65,the secretion and them RNA levels of IL- 1β and TNF- α in THP- 1 cells induced by hypoxia（P〈0.05）. Conclusion Hypoxia can induce theexpression of inflammatory cytokines,which may be related to the activation of JAK2/STAT3 signaling pathway.
出处
《中国热带医学》
CAS
2017年第2期118-122,共5页
China Tropical Medicine
基金
国家自然科学基金(No.81372977)
