核转录因子-κB信号途径对鼻息肉细胞中缺氧诱导因子-1α和血管内皮生长因子表达的影响

摘要

目的 观察核转录因子-κB (NF-κB)信号途径对低氧培养条件下鼻息肉细胞中缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)表达的影响,探讨NF-κB信号途径与鼻息肉发生、发展的关系.方法 收集2012年1月至2014年12月佳木斯大学附属第一医院耳鼻喉手术切除的鼻息肉及下鼻甲组织标本,取鼻息肉和下鼻甲组织,获得鼻息肉细胞和下鼻甲细胞,进行细胞原代培养,待细胞生长至90％时进行低氧培养;另取体外培养生长至90％的细胞,加入NF-κB抑制剂BAY11-7082(抑制剂处理组),不加抑制剂的细胞作为对照(未加抑制剂组),然后进行低氧培养;分别收取培养0、3、6、9h的细胞,采用Western blot法检测细胞中HIF-1α、VEGF、NF-κB p65蛋白的表达.结果 与0h时比较,低氧培养3、6、9h时鼻息肉细胞中HIF-1α、VEGF、NF-κB p65蛋白表达均显著增加(P＜0.05),但下鼻甲细胞中HIF-1α、VEGF、NF-κB p65蛋白表达差异均无统计学意义(P＞0.05).低氧培养6h时鼻息肉细胞中HIF-1α、VEGF、NF-κB p65蛋白表达显著高于低氧培养3、9h时(P＜0.05),但低氧培养3h与9h时鼻息肉细胞中HIF-1α、VEGF、NF-κB p65蛋白表达比较差异均无统计学意义(P＞0.05).与0h时比较,低氧培养3、6、9h时未加抑制剂组鼻息肉细胞中HIF-1α、VEGF蛋白表达均显著增加(P＜0.05);低氧培养6h时未加抑制剂组鼻息肉细胞中HIF-1α、VEGF蛋白表达显著高于低氧培养3、9h时(P＜0.05),但低氧培养3h与9h时未加抑制剂组鼻息肉细胞中HIF-1α、VEGF蛋白表达比较差异均无统计学意义(P＞0.05).低氧培养0、3、6、9h时抑制剂处理组鼻息肉细胞中HIF-1α、VEGF蛋白表达比较差异均无统计学意义(P＞0.05).低氧培养0h时,未加抑制剂组与抑制剂处理组鼻息肉细胞中HIF-1α和VEGF蛋白表达比较差异均无统计学意义(P＞0.05).低氧培养3、6、9h时,抑制剂处理组鼻息肉细胞中HIF-1α和VEGF蛋白表达均显著低于未加抑制剂组(P＜0.05).结论 在低氧条件下,鼻息肉细胞中HIF-1α、VEGF、NF-κB p65蛋白表达增加;NF-κB信号通路可能介导了低氧诱导HIF-1α和VEGF蛋白表达的过程,进而参与鼻息肉的发生和发展.%Objective To observe the effect of nuclear factor-κB (NF-κB) signaling pathway on the expression of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in nasal polyp cells under hypoxic cultivation,and to investigate the relationship between NF-κB signaling pathway and the development of nasal polyp.Methods The nasal polyp and inferior turbinate tissue specimens were collected in the First Affiliated Hospital of Jiamusi University from January 2012 to December 2014.The nasal polyp and inferior turbinate tissues were taken to obtain nasal polyp cells and inferior turbinate cells,then the cells were cultured in primary culture,and the cells were cultured under hypoxia when they grew to 90％.When the cells were cultured in vitro to 90％,the NF-κB inhibitor BAY11-7082 was added (inhibitor intervention group),the other cells without inhibitor were used as controls (no inhibitor group),then the cells in the two groups were cultured under hypoxia.The cells were collected when they were cultured for 0,3,6 and 9 hours,respectively;and the expression of HIF-1α,VEGF and NF-κB p65 protein in the cells were detected by Western blot.Results Compared with 0 hour,the expression of HIF-1α,VEGF and NF-κB p65 protein in nasal polyp cells increased significantly after 3,6 and 9 hours of hypoxic cultivation (P ＜ 0.05);however,the expression of HIF-1α,VEGF and NF-κB p65 protein in inferior turbinate cells was not statistically significant (P ＞ 0.05).The expression of HIF-1α,VEGF and NF-κB p65 protein in nasal polyposis cells after 6 hours of hypoxic cultivation was significantly higher than that after 3 and 9 hours of hypoxic cultivation (P ＜ 0.05);but there was no significant difference in the expression of HIF-1α,VEGF and NF-κB p65 protein in nasal polyp cells between 3 and 9 hours of hypoxic cultivation (P ＞ 0.05).Compared with 0 hour,the expression of HIF-1α and VEGF protein in nasal polyp cells of no inhibitor group increased significantly after 3,6 and 9 hours of hypoxic cultivation (P ＜ 0.05);and the expression of HIF-1α and VEGF protein in nasal polyp cells after 6 hours of hypoxic cultivation was significantly higher than that after 3 and 9 hours of hypoxic cultivation in no inhibitor group (P ＜ 0.05).But there was no significant difference in the expression of HIF-1α and VEGF protein in nasal polyp cells of no inhibitor group between 3 and 9 hours of hypoxic cultivation (P ＞ 0.05).There was no significant difference in the expression of HIF-1 α and VEGF protein in nasal polyp cells of the inhibitor intervention group among 0,3,6 and 9 hours of hypoxic cultivation (P ＞ 0.05).There was no significant difference in the expression of HIF-1α and VEGF protein in nasal polyp cells between no inhibitor group and inhibitor intervention group at 0 hour of hypoxic cultivation (P ＞0.05).The expression of HIF-1α and VEGF protein in nasal polyp cells of inhibitor intervention group was significantly lower than that of no inhibitor group after 3,6 and 9 hours of hypoxic cultivation (P ＜ 0.05).Conclusion The expression of HIF-1α,VEGF and NF-κB p65 protein increased in nasal polyp cells under hypoxia condition.NF-κB signaling pathway may mediate hypoxia-induced HIF-1α and VEGF protein expression,and participate in the occurrence and development of nasal polyp.