PEDF对低氧状态下人支气管上皮细胞表达VEGF的影响

发布时间：2019-04-03 16:17

【摘要】：目的:研究色素上皮衍生因子(PEDF)对低氧状态下人支气管上皮细胞(HBE)分泌血管内皮生长因子(VEGF)的影响,并探讨该作用与缺氧诱导因子1α(HIF-1α)的相关性。方法:体外培养的HBE细胞取第3~7代用于实验,氯化钴(CoCl2)100μmol/L模拟缺氧状态。实验分为4组:正常对照组(A组)、氯化钴干预组(B组)、氯化钴+PEDF 50 ng/ml干预组(C组)及氯化钴+PEDF 200 ng/ml干预组(D组)。RT-PCR法检测各组细胞HIF-1α及VEGF在mRNA水平上的表达差异;ELISA及Western blot法检测各组细胞VEGF蛋白表达水平;细胞免疫荧光染色法(IF)检测各组细胞HIF-1α蛋白在细胞质和细胞核中的表达情况。结果:①B组VEGF的mRNA水平较A组显著升高[为A组的(8.56±0.67)倍,P0.01],C组与D组的VEGF mRNA水平较B组明显下降(P0.01);B组HIF-1αmRNA的表达水平较A组明显升高(P0.01),B、C、D 3组间HIF-1αmRNA无统计学差异;②细胞培养基上清中B组VEGF表达量[(1 370.10±42.98)pg/ml]较A组[(670.00±23.35)pg/ml]明显升高(P0.01),且B组与C组[(816.19±37.05)pg/ml]及D组[(646.47±22.70)pg/ml]比较差异有统计学意义(P0.01)。③Western blot结果显示B组VEGF的蛋白表达量为A组的(3.99±0.37)倍,差异有统计学意义(P0.01);C组与D组VEGF的蛋白表达水平较B组明显下降,差异有统计学意义(P0.05)。④IF结果显示,B组HIF-1α的表达水平及核内转移较A组和D组明显升高。结论:PEDF对低氧状态下HBE细胞高表达的VEGF具有一定抑制作用,且该抑制作用可能与PEDF调控HIF-1α有关。
[Abstract]:Aim: to study the effect of pigment epithelium derived factor (PEDF) on the secretion of vascular endothelial growth factor (VEGF) (VEGF) by human bronchial epithelial cells (HBE) in hypoxic condition, and to explore the correlation between the effect and hypoxia inducible factor 1 伪 (HIF-1 伪). Methods: the third to seventh passages of HBE cells were cultured in vitro. Cobalt chloride (CoCl2) 100 渭 mol / L was used to simulate hypoxia. The experiment was divided into four groups: normal control group (group A), cobalt chloride intervention group (group B). The expression of HIF-1 伪 and VEGF in the cells of the cobalt chloride PEDF 50 ng/ml intervention group (group C) and cobalt chloride PEDF 200 ng/ml intervention group (group D) were detected by RT-PCR method. The expression of HIF-1 伪 and VEGF in the cells of each group were measured at the level of mRNA. The expression of VEGF protein was detected by ELISA and Western blot, and the expression of HIF-1 伪 protein in cytoplasm and nucleus was detected by immunofluorescence staining (IF). Results: the mRNA level of VEGF in group 1B was significantly higher than that in group A [(8.56 卤0.67) times than that in group A, P0.01], and the level of VEGF mRNA in groups C and D was significantly lower than that in group B (P0.01). The expression of HIF- 1 伪 mRNA in group B was significantly higher than that in group A (P0.01). There was no significant difference in HIF-1 伪 mRNA between B, C and D groups. 2 the expression of VEGF in group B [(1 370.10 卤42.98) pg/ml] was significantly higher than that in group A [(670.00 卤23.35) pg/ml] (P0.01). The expression of VEGF protein in group B was significantly higher than that in group C [(816.19 卤37.05) pg/ml] and group D [(646.47 卤22.70) pg/ml] (P0.01). The results of 3Western blot showed that the expression of VEGF in group B was higher than that in group A. 3.99 卤0.37 times, The difference was statistically significant (P0.01); The expression level of VEGF protein in group C and group D was significantly lower than that in group B (P0.05) .4if results showed that the expression level of HIF-1 伪 and nuclear metastasis in group B were significantly higher than those in group A and group D. Conclusion: PEDF can inhibit the high expression of VEGF in HBE cells under hypoxia, which may be related to the regulation of HIF-1 伪 by PEDF.
【作者单位】：南京医科大学第一附属医院呼吸内科;
【基金】：江苏高校优势学科建设工程项目
【分类号】：R562.25

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