论文部分内容阅读
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 observe whether hypoxia can promote the secretion of human monocyte inflammatory cytokines and explore whether it plays a role through the PI3K / Akt and hypoxia inducible factor (HIF-1α) signaling pathways. Methods 1 Human hypoxic (THP-1) cells were cultured in hypoxia (1% O2) for 24 h. 2 THP-1 cells were pretreated with PI3K inhibitor (LY294002) and HIF-1α inhibitor (KC7F2) Hypoxia intervention was performed. 3 THP-1 cells were intervened by adding Akt agonist (insulin) and HIF-1α agonist (dimethoxyglycylglycine, DMOG) under normoxia respectively. The levels of p-Akt and HIF-1αprotein were detected by Western blot. The changes of IL-1β and TNF-αlevels were detected by ELISA. Results Compared with the control group (normoxic culture), the expression of p-Akt and HIF-1α in THP-1 cells increased and the secretion of IL-1β and TNF-α increased ). Compared with hypoxia group (hypoxia group), pretreatment with PI3K inhibitor significantly decreased the expression of p-Akt and HIF-1α, while HIF-1α inhibitor pretreatment decreased HIF-1α protein expression (P <0.05). Compared with the control group, Akt agonist can increase the expression of p-Akt, HIF -1α protein expression, HIF-1α agonist can increase the protein expression of HIF-1α, but does not affect the expression of p-Akt, both drugs can increase the secretion of IL-1β and TNF-α, the difference was statistically significant Significance (P <0.05). Conclusion Hypoxia may promote the development of chronic inflammation by activating the PI3K / Akt / HIF-1α signaling pathway and promoting the secretion of inflammatory cytokines IL-1β and TNF-α in THP-1 cells.