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  本文選題：海水　切入點：肺泡上皮細胞　出處：《中國人民解放軍醫(yī)學院》2013年碩士論文　論文類型：學位論文

【摘要】：目的: 建立海水淹溺肺損傷的細胞模型，通過觀察海水干預后肺泡上皮細胞HIF-1α蛋白的表達變化及其與細胞損傷程度、炎性因子表達的相互關系，，初步探討HIF-1α在SWD-ALI中的可能作用。 方法: 肺泡上皮細胞來源的A549細胞系，分為正常對照組(C)和海水處理組(S)。C組用新鮮培養(yǎng)基常規(guī)培養(yǎng)，S組經滅菌配方海水孵育0.5h、1h、2h、4h、8h。 1、光鏡觀察瑞氏-吉姆薩染色后細胞形態(tài)變化 2、MTT檢測細胞增殖 3、蛋白免疫印記法檢測各組細胞HIF-1α蛋白的表達 4、放射免疫法檢測細胞培養(yǎng)上清液中TNF-α、IL-6的含量 結果： 1、海水處理后A549細胞失去正常長梭形或多角形形態(tài)，細胞變圓、胞體變小，部分細胞出現核濃縮、深染、核碎裂表現。 2、與對照組相比，海水處理組A549細胞的生長曲線發(fā)生變化，海水干預0.5h、1h組細胞生長抑制率較對照組比較無統(tǒng)計學差異(P0.05)，海水干預2h、4h、8h組細胞生長抑制率較對照組比較有統(tǒng)計學差異(P0.01)；細胞生長至48h和72h時，海水干預4h組細胞生長抑制率較2h組有統(tǒng)計學差異(P0.05)，海水干預8h組細胞生長抑制率較4h組有統(tǒng)計學差異(P0.01)。 3、與對照組相比，HIF-1α在海水干預1h后開始升高，2h時升高明顯(P0.01)，4h達高峰(P＜0.01)，此后HIF-1α蛋白表達逐漸下降，但仍顯著高于對照組(P＜0.01)。 4、TNF-α與IL-6在海水干預1h后開始升高，2h達高峰(P＜0.01)，4h及8h組較對照組比較無統(tǒng)計學差異。 結論: 1、海水干預導致肺泡上皮細胞發(fā)生了不同程度的損傷，海水長時間干預可導致肺泡上皮細胞生長受抑。 2、海水干預可誘導肺泡上皮細胞TNF-α、IL-6的表達，海水干預后肺泡上皮細胞發(fā)生了炎癥反應。 3、海水干預可誘導肺泡上皮細胞HIF-1α蛋白表達；HIF-1α及相關基因表達可能參與了海水干預所致肺泡上皮細胞損傷及炎癥反應的過程。
[Abstract]:Objective:. A cell model of lung injury after seawater drowning was established. By observing the changes of HIF-1 偽 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 偽 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 偽 protein in cells of each group was detected by protein imprinting assay. 4. Radioimmunoassay was used to detect the content of TNF- 偽 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 偽 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 偽 protein decreased gradually, but it was still significantly higher than that of the control group (P < 0.01). 4TNF- 偽 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- 偽 and IL-6 in alveolar epithelial cells, and inflammatory reaction occurred in alveolar epithelial cells after seawater intervention. 3. The expression of HIF-1 偽 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.
【學位授予單位】：中國人民解放軍醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R563.9

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