本文選題：kupffer細(xì)胞 + 肝X受體α��； 參考：《重慶醫(yī)科大學(xué)》2009年碩士論文

【摘要】： 目的:通過(guò)觀察LXRα激動(dòng)劑T0901317對(duì)LPS刺激后kupffer細(xì)胞(KCs)中IRF3及GRIP1、LXRα表達(dá)的影響,探討LXRα負(fù)性調(diào)控LPS的KCs激活所到的炎癥反應(yīng)的相關(guān)機(jī)制。 方法:采用“膠原酶原位灌注法”分離和培養(yǎng)雄性KM小鼠肝臟中的KCs,所得細(xì)胞在含20%小牛血清和1%青霉素/鏈霉素的1640培養(yǎng)基中培養(yǎng)。將分離的KCs隨機(jī)分為4組:1.空白對(duì)照組;2.TLR4配體激活劑LPS(1ug/ml)組;3.LXRα配體激動(dòng)劑T0901317(5ug/ml)組;4.LPS和T0901317聯(lián)合處理組。收集培養(yǎng)細(xì)胞,采用Western bolt法檢測(cè)KCs的LXRα、GRIP1及IRF3蛋白表達(dá)水平。酶聯(lián)免疫吸附法(ELISA)檢測(cè)KCs培養(yǎng)上清液中IFNβ、TNF-α和IL-1β含量。 結(jié)果:LXRα蛋白質(zhì)表達(dá)水平在T0901317處理組(0.654±0.004)最高,LPS處理組(0.008±0.003)最低。聯(lián)合處理組(0.123±0.011)中,LXRα表達(dá)明顯低于LXRα激動(dòng)劑組(P0.05),但又顯著高于其它兩組(P0.05)。IRF3及GRIP1蛋白表達(dá)量在LPS組最高(分別為0.977±0.019, 0.761±0.005 ),聯(lián)合處理組表達(dá)明顯降低(分別為0.908±0.011, 0.710±0.003),兩組間均數(shù)差異均有顯著意義(P0.05);在LPS組及聯(lián)合處理組中,IRF3及GRIP1蛋白表達(dá)量均高于對(duì)照組(分別為0.719±0.064、0.492±0.023)及LXRα激動(dòng)劑組(分別為0.468±0.019、0.425±0.002),差別具有統(tǒng)計(jì)學(xué)意義(P0.05)。IFNβ在LPS處理組的含量(329.5±35)較對(duì)照組(129.6±17)及LXRα激動(dòng)劑組(112.8±24)有明顯增高,具有顯著性差異(P0.05);聯(lián)合處理組IFNβ含量(224.4±33)比LPS處理組明顯降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05);IFNβ表達(dá)LXRα激動(dòng)劑組表達(dá)最低。TNF-α在LPS處理組的含量(15.45±0.59)較其它三組明顯增高,具有顯著性差異(P0.05);對(duì)照組(5.05±0.17)、LXR激動(dòng)劑T0901317處理組(5.67±0.25)和聯(lián)合處理組(6.62±0.38)水平較低,且他們?nèi)M之間沒(méi)有明顯差別(P0.05)。IL-1β的表達(dá)趨勢(shì)與TNF-α相同。 結(jié)論:預(yù)防性應(yīng)用LXRα激動(dòng)劑T0901317,能明顯抑制LPS誘導(dǎo)的KCs的IRF3及GRIP1表達(dá),通過(guò)抑制IRF3途徑而發(fā)揮抗炎效應(yīng),從而抑制LPS所誘導(dǎo)的KCs活化。
[Abstract]:Aim: to investigate the effect of LXR 偽 agonist T0901317 on the expression of IRF3 and GRIP1pLXR 偽 in kupffer cells stimulated by LPS, and to explore the mechanism of LXR 偽 negatively regulating the KCs activation of LPS. Methods: KCs in the liver of male km mice were isolated and cultured by collagenase in situ perfusion method. The cells were cultured in 1640 medium containing 20% calf serum and 1% penicillin / streptomycin. The separated KCs was randomly divided into 4 groups: 1. LXR 偽 ligand agonist T09013175ugp / ml group was treated with LPS and T0901317. The expression of LXR 偽 GRIP1 and IRF3 protein in KCs were detected by Western bolt assay. The content of IFN 尾 -TNF- 偽 and IL-1 尾 in supernatant of KCs culture was determined by Elisa. Results the protein expression level of 1: LXR 偽 was the lowest in T0901317 treatment group (0.654 鹵0.004) and LPS treatment group (0.008 鹵0.003). The expression of LXR 偽 in the combined treatment group was significantly lower than that in the LXR 偽 agonist group, but it was significantly higher than that in the other two groups (0.977 鹵0.019, 0.761 鹵0.005, 0.908 鹵0.011, 0.710 鹵0.003, respectively) in the LPS group, and the highest in the LPS group (0.977 鹵0.019, 0.761 鹵0.005, respectively). The expression of LXR 偽 in the combined treatment group was significantly lower than that in the other two groups (0.908 鹵0.011, 0.710 鹵0.003, respectively). The expression of IRF3 and GRIP1 protein in LPS group and combined treatment group were significantly higher than those in control group (0.719 鹵0.064 鹵0.492 鹵0.023) and LXR 偽 agonist group (0.468 鹵0.019 鹵0.425 鹵0.002, respectively). Compared with the control group (129.6 鹵17) and the LXR 偽 agonist group (112.8 鹵24), it was significantly higher than that of the control group. The content of IFN 尾 in the combined treatment group was significantly lower than that in the LPS group, and the lowest expression of LXR 偽 agonist group in the combined treatment group was 15.45 鹵0.59, which was significantly higher than that in the other three groups. There was a significant difference in P0.05, and the levels of T0901317 agonist T0901317 in the control group and in the combined treatment group were 5.67 鹵0.25 and 6.62 鹵0.38, respectively, and there was no significant difference between the three groups in the expression trend of P0.05U. IL-1 尾 and that of TNF- 偽 in the control group (5.05 鹵0.17) and the combined treatment group (6.62 鹵0.38). Conclusion: prophylactic application of LXR 偽 agonist T0901317 can significantly inhibit the expression of IRF3 and GRIP1 in KCs induced by LPS, play an anti-inflammatory effect by inhibiting IRF3 pathway, and thus inhibit KCs activation induced by LPS.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R362

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 ;In vitro expression of CD14 protein and its gene in Kupffer cells induced by lipopolysaccharide[J];Hepatobiliary & Pancreatic Diseases International;2003年04期

2 ;The U937 cell line induced to express CD14 protein by 1,25-dihydroxyvitamin D3 and be sensitive to endotoxin stimulation[J];Hepatobiliary & Pancreatic Diseases International;2005年01期

3 ;Glycine blunts transplantative liver ischemia-reperfusion injury by downregulating interleukin 1 receptor associated kinase-4[J];Acta Pharmacologica Sinica;2006年11期

4 ;Intestinal damage mediated by Kupffer cells in rats with endotoxemia[J];World Journal of Gastroenterology;2002年05期

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本文編號(hào)：1938085