天然免疫细胞在酒精性肝病中的作用机制研究
发布时间：2018-11-02 10:28
【摘要】：长期大量饮酒可导致酒精性脂肪肝的形成,进而发展成为酒精性肝炎、肝纤维化、肝硬化乃至肝癌等一系列酒精性肝病(ALD)。酒精性肝病的形成机制是综合而且多因素的,涉及到包括多种细胞、细胞因子、补体成分及转录因子等在内的许多成分和信号分子之间的相互作用和反馈交谈。肝脏天然免疫不仅在抵抗病原体及肿瘤转化的天然防御中起到重要作用,而且参与调节肝损伤、修复肝纤维化。在酒精性肝病的发展过程中,天然免疫细胞,尤其是kuffer细胞及NK细胞在此过程中发挥着重要作用。酒精可以通过促进肝脏kuffer细胞产生ROS及促炎性因子TNF-a,同时抑制NK/IFN-γ的抗纤维化活性,来促进肝脏炎症及纤维化的发生。然而,在肝脏天然免疫中占据重要地位并且作为联系天然免疫与获得性免疫桥梁的.NKT细胞,在酒精性肝病发生发展过程中扮演的角色尚不清楚。本研究通过建立chronic-binge酒精饲喂小鼠模型,初步探究了NKT细胞对酒精性肝病发生发展的重要影响,及其与Kupffer细胞、NK细胞间的相互作用机制。 取得的结果主要有以下两个方面： 1.Kupffer细胞通过产生IL-1β招募并活化iNKT细胞从而促进酒精性肝病的发生发展。 我们发现,在chronic-binge模型中,小鼠肝脏呈现显著脂肪变及病理损伤的同时,伴随着iNKT细胞比例及绝对数的大幅升高,缺失iNKT细胞的Jα18KO小鼠肝脏损伤及脂肪变程度则明显减轻。采用IL-1Ra处理、氯磷酸盐脂质体清除Kupffer细胞以及采用纳米颗粒包被的siRNA特异性干扰Kupffer细胞中IL-1β的表达,均可以有效抑制NKT细胞向肝脏的聚集及其活化,同时减轻小鼠肝脏脂肪变及病理损伤。而给小鼠肝实质细胞中过表达IL-1β,则可以从一定程度上补偿Kupffer细胞清除所带来的影响。进一步研究发现,酒精饲喂可以上调小鼠Kupffer细胞中成熟形式的IL-1β的表达,同时伴随着NLRP3炎性小体的组分,包括NLRP3、ASC及caspase-1表达的升高,而NLRP3的缺失也具有与Kupffer细胞清除同样的效果,即抑制NKT细胞向肝脏的聚集和减轻小鼠肝脏脂肪化及病理损伤。 结论：酒精诱导的Kupffer细胞中NLRP3炎性小体的活化导致下游IL-1β的产生,招募并活化肝脏iNKT细胞,进而诱导酒精性肝损伤。 2. iNKT细胞通过产生IL-10抑制NK细胞脱颗粒及IFN-y的分泌促进酒精性肝病的发生发展。 在chronic-binge模型中,小鼠肝脏NK细胞的数量随灌胃后时间的延长逐渐下降,与NKT细胞呈现相反的动力学变化。iNKT细胞缺失的Jα18KO小鼠中,灌胃后9小时肝脏NK细胞数量明显增加,脱颗粒及EFN-γ分泌能力显著增强,同时伴随着减轻的肝脏脂肪化及损伤,而采用AsGM1清除Jα18KO小鼠的NK细胞则可以加重其肝脏脂肪变及损伤。通过转输iNKT细胞给Jα18KO鼠,以及分别转输WT小鼠及Jα18KO小鼠的MNCs给Rag1KO鼠,进一步确认了iNKT细胞通过抑制NK细胞的数量和功能发挥促进小鼠肝脏脂肪变及病理损伤的重要作用。另外,机制研究表明,iNKT细胞分泌的IL-10是抑制NK细胞功能的主要介质。使用IL-10处理Jα18KO鼠,NK细胞的数量和功能受到抑制,而在IL-10KO鼠中,被抑制的NK细胞的数量和功能得到恢复。 结论：酒精饲喂导致的肝脏中聚集并活化的iNKT细胞,可以通过产生抑制性细胞因子IL-10,抑制NK细胞的数量及功能,进而促进酒精性脂肪肝炎的发病进程。 综上所述,酒精摄入导致Kupffer细胞中NLRP3炎性小体的活化,其下游产物IL-1β进而招募并活化肝脏iNKT细胞。肝脏中大量聚集的iNKT细胞通过产生IL-10,进一步抑制NK细胞的数量及功能,从而促进酒精性脂肪变及肝损伤的发生。
[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.
【学位授予单位】：中国科学技术大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R575
本文编号：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.
【学位授予单位】：中国科学技术大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R575
【参考文献】
相关期刊论文 前3条
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
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