摘要
目的观察低氧是否促进人单核细胞炎症因子的分泌,以及探讨其是否通过PI3K/Akt和低氧诱导因子(HIF-1α)信号通路而发挥作用。方法 1体外低氧(1%O2)培养人源单核细胞系THP-1,24 h;2分别加入PI3K抑制剂(LY294002)和HIF-1α抑制剂(KC7F2)预处理THP-1细胞后再进行低氧干预;3在常氧下,分别加入Akt激动剂(胰岛素)和HIF-1α激动剂(二甲氧乙二酰甘氨酸,DMOG)对THP-1细胞进行干预。Western blot检测p-Akt和HIF-1α蛋白表达的水平,ELISA法检测IL-1β和TNF-α分泌的变化。结果与对照组(常氧培养)比较,低氧干预后THP-1细胞p-Akt、HIF-1α的蛋白表达增加,IL-1β、TNF-α分泌增加,差异有统计学意义(P<0.05);与低氧组(低氧培养)比较,PI3K抑制剂预处理能显著降低p-Akt、HIF-1α的蛋白表达,HIF-1α抑制剂预处理能降低HIF-1α的蛋白表达,但不影响p-Akt表达水平,两种药物预处理都能减少IL-1β和TNF-α分泌,差异有统计学意义(P<0.05);与对照组比较,Akt激动剂能增加p-Akt、HIF-1α的蛋白表达,HIF-1α激动剂能增加HIF-1α的蛋白表达,但不影响p-Akt表达水平,两种药物预处理都能增加IL-1β和TNF-α分泌,差异有统计学意义(P<0.05)。结论低氧可能是通过激活PI3K/Akt/HIF-1α信号通路,促进THP-1细胞炎症因子IL-1β和TNF-α的分泌,从而促进慢性炎症的发展。
Objective To investigate whether hypoxia promotes the secretion of inflammation cytokines,and whether viaPI3K/Akt and hypoxia inducible factor-1（HIF-1α）pathways in human monocytic cell line THP-1. Methods 1THP-1cells were exposed to hypoxic condition（1% O2）for 24 h;2The cells were pretreated with PI3 K inhibitor（LY294002）and HIF-1α inhibitor（KC7F2）respectively,and then exposed to hypoxia condition; 3 After the cells were treated with Akt activator（insulin）and HIF-1α activator（dimethyloxalyl glycine,DMOG）respectively,the cells were cultured under normoxia. Westernblotting was used to detect the expression levels of p-Akt and HIF-1α in above mentioned groups of cells. The concentrationsof IL-1β and TNF-α in the supernatant were determined by ELISA. Results Hypoxic exposure resulted in significantlyincreased expression levels of p- Akt and HIF- 1α,and obviously elevated concentrations of IL- 1β and TNF- α in thesupernatant compared with the cells under normoxia（P〈0.05）. However,LY294002 significantly inhibited the expression of p-Akt and HIF-1α in the cells after hypoxic exposure,while KC7F2 only decreased the protein level of HIF-1α,but had noeffect on that of p-Akt. Both LY294002 and KC7F2 pretreatment could significantly decrease the secretion of IL-1β and TNF-α after hypoxia（P〈0.05）. Compared with the control group,insulin stimulation increased the protein levels of p-Akt and HIF-1α,but DMOG enhanced HIF- 1α protein level,and exerted no effect on p- Akt expression. Both insulin and DMOGsignificantly increased the concentrations of IL-1β and TNF-α（P〈0.05）. Conclusion Hypoxia may enhance the secretionof inflammatory cytokines IL-1β and TNF-α in THP-1 cells,through activating PI3K/Akt/HIF-1α pathway,and thus promotethe development of chronic inflammation.
出处
《中国热带医学》
CAS
2016年第5期451-454,共4页
China Tropical Medicine
基金
国家自然科学基金(No.81372977)
