本文選題：京尼平 + 炎性體�。� 參考：《吉林大學(xué)》2015年碩士論文

【摘要】：炎性體是天然免疫模式識別受體（PPPs）識別病原相關(guān)分子模式（PAMPs）或損傷相關(guān)分子模式（DAMPs）后在胞漿內(nèi)形成的一種多蛋白復(fù)合物。炎性體主要由模式識別受體、ASC和caspase-1構(gòu)成，在病原感染、炎癥以及自身免疫性疾病等過程中發(fā)揮重要作用�，F(xiàn)已發(fā)現(xiàn)多種炎性體，尤以對NLRC4、NLRP3、AIM2等炎性體的研究較為深入，，但對其具體的活化和負調(diào)控機制仍不十分清楚，亟待進一步闡明。 京尼平（Genipin）是梔子苷經(jīng)β-葡萄糖苷酶水解后的產(chǎn)物，廣泛存在于梔子、杜仲等植物中。已有研究表明，京尼平在抗感染、抗炎以及治療肝臟疾病和Ⅱ型糖尿病等自身免疫性疾病過程中具有一定的效果。盡管對于京尼平的功效及機制有一定認識，但其調(diào)節(jié)炎癥反應(yīng)的機制并不十分清楚，因而限制了京尼平在疾病治療中的應(yīng)用。京尼平是否參與調(diào)控炎性體信號，進而對其所介導(dǎo)的炎癥具有調(diào)節(jié)作用，是本課題熱切關(guān)注的科學(xué)問題。 有研究表明，在綠膿假單胞桿菌、軍團菌、肺炎克雷伯菌等病原感染過程中通過活化NLRC4炎性體，誘導(dǎo)肺炎的發(fā)生。因此，本課題在體實驗以沙門氏菌重組鞭毛蛋白誘導(dǎo)的小鼠肺炎模型為研究對象，探究京尼平對NLRC4炎性體介導(dǎo)的炎癥結(jié)局的影響。用ELISA和免疫組織化學(xué)方法檢測肺部炎性因子分泌和炎性細胞浸潤，并評價京尼平對細菌鞭毛蛋白引起的肺組織病理變化的作用。結(jié)果表明，京尼平可以顯著抑制肺組織中IL-1β與KC的分泌和中性粒細胞的浸潤，同時緩解鞭毛蛋白導(dǎo)致的肺泡出血，肺泡間質(zhì)增厚。由此表明京尼平能夠顯著抑制細菌鞭毛蛋白引起的小鼠肺炎。 體外實驗以小鼠原代巨噬細胞為研究模型，探究京尼平對鼠傷寒沙門氏菌及沙門氏菌重組鞭毛蛋白誘導(dǎo)的NLRC4炎性體活化的負調(diào)控機制。應(yīng)用免疫蛋白印跡的方法檢測NLRC4、UCP2等蛋白的表達以及Caspase-1、IL-1β的活化片段和ASC寡聚化；流式細胞術(shù)檢測細胞死亡及ROS的產(chǎn)生；免疫熒光分析ASC斑點的形成和LC3蛋白的表達；ELISA檢測IL-1β和TNFα細胞因子的分泌。結(jié)果表明，京尼平顯著抑制鼠傷寒沙門氏菌及沙門氏菌重組鞭毛蛋白誘導(dǎo)的Caspase-1剪切成熟和IL-1β分泌，并且通過抑制細胞自噬以及NLRC4蛋白的表達，實現(xiàn)對NLRC4炎性體的負調(diào)控。 綜上所述，本研究確證京尼平通過抑制NLRC4蛋白的表達和細胞自噬抑制NLRC4炎性體的活化，進而有效緩解和治療沙門氏菌重組鞭毛蛋白引起的小鼠肺炎。
[Abstract]:Inflammatory body is a polyprotein complex formed in cytoplasm by innate immune pattern recognition receptors (PPPs) which recognize pathogen-associated molecular patterns (PAMPs) or damage related molecular patterns (DAMPs). Inflammatory bodies are mainly composed of pattern recognition receptors ASC and caspase-1, which play an important role in pathogenic infection, inflammation and autoimmune diseases. Many kinds of inflammatory bodies have been found, especially NLRC4NLRP3AIM2, but their specific activation and negative regulation mechanisms are still unclear and need to be further elucidated. Genipin is a product of geniposide hydrolyzed by 尾-glucosidase. It is widely found in Gardenia jasminoides and Eucommia ulmoides. Studies have shown that geniapine has a certain effect in the process of anti-infection, anti-inflammation and the treatment of autoimmune diseases such as liver disease and type II diabetes. Although there is a certain understanding of the efficacy and mechanism of geniapine, its mechanism of regulating inflammatory response is not very clear, which limits the application of genipin in the treatment of diseases. Whether geniapine is involved in regulating inflammatory signal and then regulating inflammation mediated by genistein is a scientific issue which has been paid close attention to. Some studies have shown that in the process of infection of Pseudomonas aeruginosa, Legionella pneumoniae and Klebsiella pneumoniae, pneumonia is induced by activating the inflammatory body of NLRC4. Therefore, in vivo, we investigated the effect of geniapine on inflammatory outcome mediated by NLRC4 in mice pneumonia model induced by recombinant flagellin of Salmonella. Lung inflammatory factor secretion and inflammatory cell infiltration were detected by Elisa and immunohistochemistry, and the effect of geniapine on pathological changes of lung tissue induced by bacterial flagellin was evaluated. The results showed that geniapine could significantly inhibit the secretion of IL-1 尾 and KC and the infiltration of neutrophils in lung tissue, and alleviate alveolar hemorrhage caused by flagellin and the thickening of alveolar interstitial. It is suggested that geniapine can significantly inhibit bacterial flagellin-induced pneumonia in mice. In vitro, the mouse primary macrophages were used to investigate the negative regulation mechanism of geniapine on the activation of NLRC4 inflammatory body induced by recombinant flagellin of Salmonella typhimurium and Salmonella typhimurium. Western blot was used to detect the expression of NLRC4UCP2, the activated fragment of Caspase-1 and IL-1 尾 and ASC oligodepolymerization; flow cytometry was used to detect cell death and Ros production; immunofluorescence was used to detect the formation of ASC spots and the expression of LC3 protein. The secretion of IL-1 尾 and TNF 偽 cytokines was detected by Elisa. The results showed that geniapine significantly inhibited Caspase-1 shearing maturation and IL-1 尾 secretion induced by recombinant flagellin of Salmonella typhimurium and Salmonella typhimurium, and negatively regulated the inflammatory body of NLRC4 by inhibiting autophagy and the expression of NLRC4 protein. In conclusion, this study confirmed that geniapine can effectively relieve and treat pneumonia induced by recombinant flagellin of Salmonella by inhibiting the expression of NLRC4 protein and inhibiting the activation of NLRC4 inflammatory body by autophagy.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S853.7

【參考文獻】

相關(guān)期刊論文 前1條

1 Tai-Xiang Liu;Xin Luo;Yu-Wei Gu;Bin Yang;Zheng Wang;;Correlation of discoloration and biomechanical properties in porcine sclera induced by genipin[J];International Journal of Ophthalmology;2014年04期

本文編號：2105955