【摘要】：目的研究轉(zhuǎn)染窖蛋白1（Caveolin-1，CAV-1）基因?qū)χ嗵牵↙ipopolysaccharide，LPS）誘導(dǎo)的人支氣管上皮細(xì)胞（Humanbronchialepithelialcells，HBE）Toll樣受體4（Toll-likereceptor4，TLR-4）表達(dá)的影響，以及大黃素(Emodin)對LPS誘導(dǎo)的HBE細(xì)胞CAV-1和TLR-4表達(dá)的干預(yù)效應(yīng)。 方法本實驗分為兩部分：第一部分，對LPS誘導(dǎo)的HBE細(xì)胞給予不同濃度的Emodin干預(yù)，并設(shè)立正常對照組、單純LPS干預(yù)組、陽性對照組（Atorvastatin,阿托伐他汀）和陰性對照組（溶媒DMSO），用免疫印記法（Westernblot）檢測并比較各組細(xì)胞CAV-1的表達(dá)水平；第二部分，構(gòu)建Cav-EGFP重組質(zhì)粒，瞬時轉(zhuǎn)染HBE細(xì)胞，設(shè)立正常對照組和空質(zhì)粒轉(zhuǎn)染組，Westernblot檢測并比較三組細(xì)胞CAV-1的表達(dá)；于質(zhì)粒轉(zhuǎn)染HBE細(xì)胞48h后給予LPS刺激，Westernblot檢測并比較CAV-1過表達(dá)組與空質(zhì)粒轉(zhuǎn)染組TLR-4的表達(dá)，并給予Emodin干預(yù)轉(zhuǎn)染后LPS誘導(dǎo)的HBE細(xì)胞，Westernblot檢測比較各組細(xì)胞TLR-4的表達(dá)。 結(jié)果第一部分：與正常對照組相比，單純LPS干預(yù)組HBE細(xì)胞經(jīng)LPS刺激后，CAV-1蛋白的表達(dá)量明顯上調(diào)（P0.05)；中、高劑量Emodin干預(yù)組、陽性對照組與單純LPS干預(yù)組相比,CAV-1蛋白的表達(dá)量均明顯下降（P0.05）；第二部分：CAV-1-EGFP重組質(zhì)粒轉(zhuǎn)染組與正常對照組、空質(zhì)粒轉(zhuǎn)染組比較，CAV-1的表達(dá)量明顯上調(diào)（P0.05）,而空質(zhì)粒轉(zhuǎn)染組與正常對照組相比，，CAV-1的表達(dá)量無統(tǒng)計差異（P0.05）；與空質(zhì)粒轉(zhuǎn)染組相比，CAV-1過表達(dá)組能上調(diào)LPS誘導(dǎo)的HBE細(xì)胞TLR-4的表達(dá),Emodin能抑制該效應(yīng)(P0.05)。結(jié)論Emodin能抑制LPS誘導(dǎo)的HBE細(xì)胞CAV-1的表達(dá)；轉(zhuǎn)染CAV-1基因能 上調(diào)LPS誘導(dǎo)的HBE細(xì)胞TLR-4的表達(dá)，Emodin能抑制該效應(yīng)。
[Abstract]:Objective to study the effect of cellar protein 1 (Caveolin-1,CAV-1) gene transfection on the expression of Toll like receptor 4 (Toll-likereceptor4,TLR-4) in human bronchial epithelial cells (Humanbronchialepithelialcells,HBE) induced by lipopolysaccharide (Lipopolysaccharide,LPS) and the effect of emodin (Emodin) on LPS induced CAV-1 and TLR-4 expression in HBE cells. Methods the experiment was divided into two parts: in the first part, the HBE cells induced by LPS were treated with different concentrations of Emodin, and the normal control group was set up, and the control group was treated with LPS alone. The positive control group (Atorvastatin, Atto vastatin) and the negative control group (the solvent DMSO), was used to detect and compare the expression of CAV-1 by (Westernblot). The expression of CAV-1 was detected by Western blot in the normal control group and the empty plasmid transfection group, and the expression of TLR-4 in the CAV-1 overexpression group was detected and compared with that in the empty plasmid transfection group after 48 hours of transfection of the plasmid into HBE cells. The expression of TLR-4 was detected by Western blot in HBE cells induced by LPS after Emodin intervention. Results: the first part: compared with the normal control group, the expression of CAV-1 protein in HBE cells stimulated by LPS was significantly up-regulated in the LPS intervention group (P0.05), while in the middle and high dose Emodin intervention group, the expression of CAV-1 protein was significantly increased in the HBE cells treated with LPS (P0.05). The expression of CAV-1 protein in the positive control group was significantly lower than that in the control group (P0.05). The expression of CAV-1 was significantly up-regulated in the empty plasmid transfection group (P0.05), but there was no statistical difference between the empty plasmid transfection group and the normal control group (P0.05). Compared with the empty plasmid transfection group, the expression of TLR-4 in HBE cells induced by LPS could be upregulated in the overexpression group of CAV-1. Emodin could inhibit this effect (P0.05). Conclusion Emodin can inhibit the expression of CAV-1 in HBE cells induced by LPS, and transfection of CAV-1 gene can up-regulate the expression of TLR-4 in HBE cells induced by LPS.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R563.8

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