本文選題：海水　切入點(diǎn)：肺泡上皮細(xì)胞　出處：《中國(guó)人民解放軍醫(yī)學(xué)院》2013年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的: 建立海水淹溺肺損傷的細(xì)胞模型，通過(guò)觀察海水干預(yù)后肺泡上皮細(xì)胞HIF-1α蛋白的表達(dá)變化及其與細(xì)胞損傷程度、炎性因子表達(dá)的相互關(guān)系，，初步探討HIF-1α在SWD-ALI中的可能作用。 方法: 肺泡上皮細(xì)胞來(lái)源的A549細(xì)胞系，分為正常對(duì)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(jīng)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(xì)胞形態(tài)變化 2、MTT檢測(cè)細(xì)胞增殖 3、蛋白免疫印記法檢測(cè)各組細(xì)胞HIF-1α蛋白的表達(dá) 4、放射免疫法檢測(cè)細(xì)胞培養(yǎng)上清液中TNF-α、IL-6的含量 結(jié)果： 1、海水處理后A549細(xì)胞失去正常長(zhǎng)梭形或多角形形態(tài)，細(xì)胞變圓、胞體變小，部分細(xì)胞出現(xiàn)核濃縮、深染、核碎裂表現(xiàn)。 2、與對(duì)照組相比，海水處理組A549細(xì)胞的生長(zhǎng)曲線發(fā)生變化，海水干預(yù)0.5h、1h組細(xì)胞生長(zhǎng)抑制率較對(duì)照組比較無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)，海水干預(yù)2h、4h、8h組細(xì)胞生長(zhǎng)抑制率較對(duì)照組比較有統(tǒng)計(jì)學(xué)差異(P0.01)；細(xì)胞生長(zhǎng)至48h和72h時(shí)，海水干預(yù)4h組細(xì)胞生長(zhǎng)抑制率較2h組有統(tǒng)計(jì)學(xué)差異(P0.05)，海水干預(yù)8h組細(xì)胞生長(zhǎng)抑制率較4h組有統(tǒng)計(jì)學(xué)差異(P0.01)。 3、與對(duì)照組相比，HIF-1α在海水干預(yù)1h后開(kāi)始升高，2h時(shí)升高明顯(P0.01)，4h達(dá)高峰(P＜0.01)，此后HIF-1α蛋白表達(dá)逐漸下降，但仍顯著高于對(duì)照組(P＜0.01)。 4、TNF-α與IL-6在海水干預(yù)1h后開(kāi)始升高，2h達(dá)高峰(P＜0.01)，4h及8h組較對(duì)照組比較無(wú)統(tǒng)計(jì)學(xué)差異。 結(jié)論: 1、海水干預(yù)導(dǎo)致肺泡上皮細(xì)胞發(fā)生了不同程度的損傷，海水長(zhǎng)時(shí)間干預(yù)可導(dǎo)致肺泡上皮細(xì)胞生長(zhǎng)受抑。 2、海水干預(yù)可誘導(dǎo)肺泡上皮細(xì)胞TNF-α、IL-6的表達(dá)，海水干預(yù)后肺泡上皮細(xì)胞發(fā)生了炎癥反應(yīng)。 3、海水干預(yù)可誘導(dǎo)肺泡上皮細(xì)胞HIF-1α蛋白表達(dá)；HIF-1α及相關(guān)基因表達(dá)可能參與了海水干預(yù)所致肺泡上皮細(xì)胞損傷及炎癥反應(yīng)的過(guò)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(xué)位授予單位】：中國(guó)人民解放軍醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R563.9

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