天然免疫細胞在酒精性肝病中的作用機制研究
發(fā)布時間：2018-11-02 10:28
【摘要】：長期大量飲酒可導致酒精性脂肪肝的形成,進而發(fā)展成為酒精性肝炎、肝纖維化、肝硬化乃至肝癌等一系列酒精性肝病(ALD)。酒精性肝病的形成機制是綜合而且多因素的,涉及到包括多種細胞、細胞因子、補體成分及轉(zhuǎn)錄因子等在內(nèi)的許多成分和信號分子之間的相互作用和反饋交談。肝臟天然免疫不僅在抵抗病原體及腫瘤轉(zhuǎn)化的天然防御中起到重要作用,而且參與調(diào)節(jié)肝損傷、修復肝纖維化。在酒精性肝病的發(fā)展過程中,天然免疫細胞,尤其是kuffer細胞及NK細胞在此過程中發(fā)揮著重要作用。酒精可以通過促進肝臟kuffer細胞產(chǎn)生ROS及促炎性因子TNF-a,同時抑制NK/IFN-γ的抗纖維化活性,來促進肝臟炎癥及纖維化的發(fā)生。然而,在肝臟天然免疫中占據(jù)重要地位并且作為聯(lián)系天然免疫與獲得性免疫橋梁的.NKT細胞,在酒精性肝病發(fā)生發(fā)展過程中扮演的角色尚不清楚。本研究通過建立chronic-binge酒精飼喂小鼠模型,初步探究了NKT細胞對酒精性肝病發(fā)生發(fā)展的重要影響,及其與Kupffer細胞、NK細胞間的相互作用機制。 取得的結(jié)果主要有以下兩個方面： 1.Kupffer細胞通過產(chǎn)生IL-1β招募并活化iNKT細胞從而促進酒精性肝病的發(fā)生發(fā)展。 我們發(fā)現(xiàn),在chronic-binge模型中,小鼠肝臟呈現(xiàn)顯著脂肪變及病理損傷的同時,伴隨著iNKT細胞比例及絕對數(shù)的大幅升高,缺失iNKT細胞的Jα18KO小鼠肝臟損傷及脂肪變程度則明顯減輕。采用IL-1Ra處理、氯磷酸鹽脂質(zhì)體清除Kupffer細胞以及采用納米顆粒包被的siRNA特異性干擾Kupffer細胞中IL-1β的表達,均可以有效抑制NKT細胞向肝臟的聚集及其活化,同時減輕小鼠肝臟脂肪變及病理損傷。而給小鼠肝實質(zhì)細胞中過表達IL-1β,則可以從一定程度上補償Kupffer細胞清除所帶來的影響。進一步研究發(fā)現(xiàn),酒精飼喂可以上調(diào)小鼠Kupffer細胞中成熟形式的IL-1β的表達,同時伴隨著NLRP3炎性小體的組分,包括NLRP3、ASC及caspase-1表達的升高,而NLRP3的缺失也具有與Kupffer細胞清除同樣的效果,即抑制NKT細胞向肝臟的聚集和減輕小鼠肝臟脂肪化及病理損傷。 結(jié)論：酒精誘導的Kupffer細胞中NLRP3炎性小體的活化導致下游IL-1β的產(chǎn)生,招募并活化肝臟iNKT細胞,進而誘導酒精性肝損傷。 2. iNKT細胞通過產(chǎn)生IL-10抑制NK細胞脫顆粒及IFN-y的分泌促進酒精性肝病的發(fā)生發(fā)展。 在chronic-binge模型中,小鼠肝臟NK細胞的數(shù)量隨灌胃后時間的延長逐漸下降,與NKT細胞呈現(xiàn)相反的動力學變化。iNKT細胞缺失的Jα18KO小鼠中,灌胃后9小時肝臟NK細胞數(shù)量明顯增加,脫顆粒及EFN-γ分泌能力顯著增強,同時伴隨著減輕的肝臟脂肪化及損傷,而采用AsGM1清除Jα18KO小鼠的NK細胞則可以加重其肝臟脂肪變及損傷。通過轉(zhuǎn)輸iNKT細胞給Jα18KO鼠,以及分別轉(zhuǎn)輸WT小鼠及Jα18KO小鼠的MNCs給Rag1KO鼠,進一步確認了iNKT細胞通過抑制NK細胞的數(shù)量和功能發(fā)揮促進小鼠肝臟脂肪變及病理損傷的重要作用。另外,機制研究表明,iNKT細胞分泌的IL-10是抑制NK細胞功能的主要介質(zhì)。使用IL-10處理Jα18KO鼠,NK細胞的數(shù)量和功能受到抑制,而在IL-10KO鼠中,被抑制的NK細胞的數(shù)量和功能得到恢復。 結(jié)論：酒精飼喂導致的肝臟中聚集并活化的iNKT細胞,可以通過產(chǎn)生抑制性細胞因子IL-10,抑制NK細胞的數(shù)量及功能,進而促進酒精性脂肪肝炎的發(fā)病進程。 綜上所述,酒精攝入導致Kupffer細胞中NLRP3炎性小體的活化,其下游產(chǎn)物IL-1β進而招募并活化肝臟iNKT細胞。肝臟中大量聚集的iNKT細胞通過產(chǎn)生IL-10,進一步抑制NK細胞的數(shù)量及功能,從而促進酒精性脂肪變及肝損傷的發(fā)生。
[Abstract]:Long-term drinking can lead to the formation of alcoholic fatty liver, and further develop a series of alcoholic liver diseases (ALD), such as alcoholic hepatitis, liver fibrosis, liver cirrhosis and liver cancer. The mechanism of formation of alcoholic liver disease is comprehensive and multi-factor involving interaction and feedback between many components and signal molecules, including multiple cells, cytokines, complement components, and transcription factors. The liver natural immunity plays an important role not only in resisting pathogens and natural defense of tumor transformation, but also participates in regulating liver injury and repairing liver fibrosis. During the development of alcoholic liver disease, natural immune cells, especially kuffer cells and NK cells play an important role in this process. Alcohol can promote liver inflammation and fibrosis by promoting the production of ROS and pro-inflammatory factors, TNF-a, in the liver, kupffer cells, while inhibiting the anti-fibrotic activity of NK/ IFN-KIT. However, it plays an important role in the innate immunity of the liver and serves as a bridge between natural immunity and natural immunity. The role of NKT cells in the development of alcoholic liver disease is unknown. This study was conducted to study the important effect of NKT cells on the development of alcoholic liver disease and its interaction mechanism with Kupffer cells and NK cells. The results obtained mainly include the following two Aspect: 1. Kupffer cells promote alcoholic liver disease by producing IL-1 gene and activating iNKT cells We found that in the ic-binge model, the liver of mice showed significant changes in fat and pathological damage, accompanied by a significant increase in the proportion and absolute number of iNKT cells, the damage and fat of the liver of J-type 18KO mice lacking iNKT cells. The levels of IL-1Ra and IL-1 in Kupffer cells were significantly reduced by IL-1Ra treatment. The expression of IL-1 gene in Kupffer cells by using nano-particle-coated siRNA could effectively inhibit the aggregation and activation of NKT cells to the liver while reducing the liver of mice. The expression of IL-1 in liver parenchyma cells of mice can compensate Kupffer cells to some extent. Further study found that alcohol feeding could upregulate the expression of IL-1 gene in the mature form of Kupffer cells in mice, and also accompanied with the increase of the expression of NLRP3, ASC and caspase-1, while the deletion of NLRP3 was similar to that of Kupffer. The same effect of cell removal is to inhibit the aggregation of NKT cells to the liver and to relieve the liver of mice Conclusion: The activation of NLRP3 inflammatory small body induced by alcohol in Kupffer cells induced by alcohol leads to the generation, recruitment and activation of iNKT cells in the liver. in ord to induce alcoholic liver injury. iNKT cells inhibit NK cell degranulation and IFN-y by producing IL-10. To promote the development of alcoholic liver disease, the number of NK cells in the liver of mouse liver gradually decreased with the increase of time after intragastric administration. In the absence of iNKT cells, the number of NK cells increased significantly in 9 hours after intragastric administration. AsGM1 was used to remove the N of J-type 18KO mice. K cells can increase the liver fat and damage of the cells, and further confirm that the iNKT cells are promoted by inhibiting the quantity and function of NK cells by transferring the iNKT cells to the J-type 18KO mice and MNCs respectively transferred to WT mice and J-type 18KO mice to the Rag1KO mice. In addition, the mechanism study showed that iNKT cells secrete IL -10 is the main medium for inhibiting the function of NK cells. The number and function of NK cells are inhibited using IL-10 and the number and function of NK cells are inhibited, and in the IL-10KO mice, Conclusion: The proliferation and activation of iNKT cells in the liver caused by alcohol feeding can inhibit the number of NK cells by producing inhibitory cytokine IL-10. In conclusion, alcohol intake leads to the activation of NLRP3 inflammatory small body in Kupffer cells, A large number of iNKT cells in the liver can further inhibit NK cells by producing IL-10.
【學位授予單位】：中國科學技術(shù)大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R575
本文編號：2305758
[Abstract]:Long-term drinking can lead to the formation of alcoholic fatty liver, and further develop a series of alcoholic liver diseases (ALD), such as alcoholic hepatitis, liver fibrosis, liver cirrhosis and liver cancer. The mechanism of formation of alcoholic liver disease is comprehensive and multi-factor involving interaction and feedback between many components and signal molecules, including multiple cells, cytokines, complement components, and transcription factors. The liver natural immunity plays an important role not only in resisting pathogens and natural defense of tumor transformation, but also participates in regulating liver injury and repairing liver fibrosis. During the development of alcoholic liver disease, natural immune cells, especially kuffer cells and NK cells play an important role in this process. Alcohol can promote liver inflammation and fibrosis by promoting the production of ROS and pro-inflammatory factors, TNF-a, in the liver, kupffer cells, while inhibiting the anti-fibrotic activity of NK/ IFN-KIT. However, it plays an important role in the innate immunity of the liver and serves as a bridge between natural immunity and natural immunity. The role of NKT cells in the development of alcoholic liver disease is unknown. This study was conducted to study the important effect of NKT cells on the development of alcoholic liver disease and its interaction mechanism with Kupffer cells and NK cells. The results obtained mainly include the following two Aspect: 1. Kupffer cells promote alcoholic liver disease by producing IL-1 gene and activating iNKT cells We found that in the ic-binge model, the liver of mice showed significant changes in fat and pathological damage, accompanied by a significant increase in the proportion and absolute number of iNKT cells, the damage and fat of the liver of J-type 18KO mice lacking iNKT cells. The levels of IL-1Ra and IL-1 in Kupffer cells were significantly reduced by IL-1Ra treatment. The expression of IL-1 gene in Kupffer cells by using nano-particle-coated siRNA could effectively inhibit the aggregation and activation of NKT cells to the liver while reducing the liver of mice. The expression of IL-1 in liver parenchyma cells of mice can compensate Kupffer cells to some extent. Further study found that alcohol feeding could upregulate the expression of IL-1 gene in the mature form of Kupffer cells in mice, and also accompanied with the increase of the expression of NLRP3, ASC and caspase-1, while the deletion of NLRP3 was similar to that of Kupffer. The same effect of cell removal is to inhibit the aggregation of NKT cells to the liver and to relieve the liver of mice Conclusion: The activation of NLRP3 inflammatory small body induced by alcohol in Kupffer cells induced by alcohol leads to the generation, recruitment and activation of iNKT cells in the liver. in ord to induce alcoholic liver injury. iNKT cells inhibit NK cell degranulation and IFN-y by producing IL-10. To promote the development of alcoholic liver disease, the number of NK cells in the liver of mouse liver gradually decreased with the increase of time after intragastric administration. In the absence of iNKT cells, the number of NK cells increased significantly in 9 hours after intragastric administration. AsGM1 was used to remove the N of J-type 18KO mice. K cells can increase the liver fat and damage of the cells, and further confirm that the iNKT cells are promoted by inhibiting the quantity and function of NK cells by transferring the iNKT cells to the J-type 18KO mice and MNCs respectively transferred to WT mice and J-type 18KO mice to the Rag1KO mice. In addition, the mechanism study showed that iNKT cells secrete IL -10 is the main medium for inhibiting the function of NK cells. The number and function of NK cells are inhibited using IL-10 and the number and function of NK cells are inhibited, and in the IL-10KO mice, Conclusion: The proliferation and activation of iNKT cells in the liver caused by alcohol feeding can inhibit the number of NK cells by producing inhibitory cytokine IL-10. In conclusion, alcohol intake leads to the activation of NLRP3 inflammatory small body in Kupffer cells, A large number of iNKT cells in the liver can further inhibit NK cells by producing IL-10.
【學位授予單位】：中國科學技術(shù)大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R575
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相關期刊論文 前3條
1 ;The Roles of Innate Immune Cells in Liver Injury and Regeneration[J];Cellular & Molecular Immunology;2007年04期
2 Natalia A Osna;;Hepatitis C virus and ethanol alter antigen presentation in liver cells[J];World Journal of Gastroenterology;2009年10期
3 Sebastian Mueller;Gunda Millonig;Helmut K Seitz;;Alcoholic liver disease and hepatitis C:A frequently underestimated combination[J];World Journal of Gastroenterology;2009年28期
，本文編號：2305758
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