海水干预诱导人肺泡上皮细胞低氧诱导因子-1α表达的研究

发布时间：2018-03-06 23:13

本文选题：海水　切入点：肺泡上皮细胞　出处：《中国人民解放军医学院》2013年硕士论文　论文类型：学位论文

【摘要】：目的: 建立海水淹溺肺损伤的细胞模型，通过观察海水干预后肺泡上皮细胞HIF-1α蛋白的表达变化及其与细胞损伤程度、炎性因子表达的相互关系，，初步探讨HIF-1α在SWD-ALI中的可能作用。方法: 肺泡上皮细胞来源的A549细胞系，分为正常对照组(C)和海水处理组(S)。C组用新鲜培养基常规培养，S组经灭菌配方海水孵育0.5h、1h、2h、4h、8h。 1、光镜观察瑞氏-吉姆萨染色后细胞形态变化 2、MTT检测细胞增殖 3、蛋白免疫印记法检测各组细胞HIF-1α蛋白的表达 4、放射免疫法检测细胞培养上清液中TNF-α、IL-6的含量结果： 1、海水处理后A549细胞失去正常长梭形或多角形形态，细胞变圆、胞体变小，部分细胞出现核浓缩、深染、核碎裂表现。 2、与对照组相比，海水处理组A549细胞的生长曲线发生变化，海水干预0.5h、1h组细胞生长抑制率较对照组比较无统计学差异(P0.05)，海水干预2h、4h、8h组细胞生长抑制率较对照组比较有统计学差异(P0.01)；细胞生长至48h和72h时，海水干预4h组细胞生长抑制率较2h组有统计学差异(P0.05)，海水干预8h组细胞生长抑制率较4h组有统计学差异(P0.01)。 3、与对照组相比，HIF-1α在海水干预1h后开始升高，2h时升高明显(P0.01)，4h达高峰(P＜0.01)，此后HIF-1α蛋白表达逐渐下降，但仍显著高于对照组(P＜0.01)。 4、TNF-α与IL-6在海水干预1h后开始升高，2h达高峰(P＜0.01)，4h及8h组较对照组比较无统计学差异。结论: 1、海水干预导致肺泡上皮细胞发生了不同程度的损伤，海水长时间干预可导致肺泡上皮细胞生长受抑。 2、海水干预可诱导肺泡上皮细胞TNF-α、IL-6的表达，海水干预后肺泡上皮细胞发生了炎症反应。 3、海水干预可诱导肺泡上皮细胞HIF-1α蛋白表达；HIF-1α及相关基因表达可能参与了海水干预所致肺泡上皮细胞损伤及炎症反应的过程。
[Abstract]:Objective:. A cell model of lung injury after seawater drowning was established. By observing the changes of HIF-1 伪 protein expression in alveolar epithelial cells after seawater intervention and their relationship with the degree of cell injury and the expression of inflammatory factors, the possible role of HIF-1 伪 in SWD-ALI was preliminarily investigated. Methods:. The A549 cell line derived from alveolar epithelial cells was divided into two groups: normal control group (C) and seawater treatment group (SX 路C), which were incubated with sterilizing formula seawater for 0.5h, 1h, 2h, 4h and 8h. 1. The morphological changes of cells were observed under light microscope after Ricker-Gimsa staining. MTT assay of cell proliferation. Expression of HIF-1 伪 protein in cells of each group was detected by protein imprinting assay. 4. Radioimmunoassay was used to detect the content of TNF- 伪 and IL-6 in the supernatant of cell culture. Results:. 1. After seawater treatment, A549 cells lost the normal fusiform or polygonal shape, the cells became round, the cell bodies became smaller, and some of the cells showed nuclear condensation, deep staining and nuclear fragmentation. 2Compared with the control group, the growth curve of A549 cells in seawater treatment group was changed. There was no significant difference in the inhibition rate of cell growth between the 1 h group and the control group (P 0.05), but there was a significant difference in the inhibition rate of cell growth in the 2 h ~ 4 h ~ 8 h group compared with the control group, and the cell growth rate at 48 h and 72 h after seawater treatment was significantly higher than that of the control group. The inhibition rate of cell growth in seawater group for 4 h was significantly higher than that in group 2 (P 0.05), and the inhibition rate of cell growth in group 8 h after seawater intervention was significantly different from that in group 4 h (P 0.01). 3Compared with the control group, HIF-1 伪 increased significantly after 1 h of seawater treatment and reached the peak at 4 h after seawater treatment (P < 0.01). After that, the expression of HIF-1 伪 protein decreased gradually, but it was still significantly higher than that of the control group (P < 0.01). 4TNF- 伪 and IL-6 began to rise to the peak at 2 h after seawater intervention (P < 0.01) and 8h respectively. There was no significant difference between the two groups in comparison with the control group. Conclusion:. 1. Seawater intervention resulted in different degrees of injury of alveolar epithelial cells, and the growth of alveolar epithelial cells was inhibited by seawater intervention for a long time. 2. Seawater intervention could induce the expression of TNF- 伪 and IL-6 in alveolar epithelial cells, and inflammatory reaction occurred in alveolar epithelial cells after seawater intervention. 3. The expression of HIF-1 伪 and related genes in alveolar epithelial cells induced by seawater may be involved in the process of injury and inflammation of alveolar epithelial cells induced by seawater.
【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.9

【参考文献】