LXRα通过IRF3途径负性调控LPS诱导的Kupffer细胞激活的机制研究

发布时间：2018-05-26 16:52

本文选题：kupffer细胞 + 肝X受体α　；参考：《重庆医科大学》2009年硕士论文

【摘要】： 目的:通过观察LXRα激动剂T0901317对LPS刺激后kupffer细胞(KCs)中IRF3及GRIP1、LXRα表达的影响,探讨LXRα负性调控LPS的KCs激活所到的炎症反应的相关机制。方法:采用“胶原酶原位灌注法”分离和培养雄性KM小鼠肝脏中的KCs,所得细胞在含20%小牛血清和1%青霉素/链霉素的1640培养基中培养。将分离的KCs随机分为4组:1.空白对照组;2.TLR4配体激活剂LPS(1ug/ml)组;3.LXRα配体激动剂T0901317(5ug/ml)组;4.LPS和T0901317联合处理组。收集培养细胞,采用Western bolt法检测KCs的LXRα、GRIP1及IRF3蛋白表达水平。酶联免疫吸附法(ELISA)检测KCs培养上清液中IFNβ、TNF-α和IL-1β含量。结果:LXRα蛋白质表达水平在T0901317处理组(0.654±0.004)最高,LPS处理组(0.008±0.003)最低。联合处理组(0.123±0.011)中,LXRα表达明显低于LXRα激动剂组(P0.05),但又显著高于其它两组(P0.05)。IRF3及GRIP1蛋白表达量在LPS组最高(分别为0.977±0.019, 0.761±0.005 ),联合处理组表达明显降低(分别为0.908±0.011, 0.710±0.003),两组间均数差异均有显著意义(P0.05);在LPS组及联合处理组中,IRF3及GRIP1蛋白表达量均高于对照组(分别为0.719±0.064、0.492±0.023)及LXRα激动剂组(分别为0.468±0.019、0.425±0.002),差别具有统计学意义(P0.05)。IFNβ在LPS处理组的含量(329.5±35)较对照组(129.6±17)及LXRα激动剂组(112.8±24)有明显增高,具有显著性差异(P0.05);联合处理组IFNβ含量(224.4±33)比LPS处理组明显降低,差异有统计学意义(P0.05);IFNβ表达LXRα激动剂组表达最低。TNF-α在LPS处理组的含量(15.45±0.59)较其它三组明显增高,具有显著性差异(P0.05);对照组(5.05±0.17)、LXR激动剂T0901317处理组(5.67±0.25)和联合处理组(6.62±0.38)水平较低,且他们三组之间没有明显差别(P0.05)。IL-1β的表达趋势与TNF-α相同。结论:预防性应用LXRα激动剂T0901317,能明显抑制LPS诱导的KCs的IRF3及GRIP1表达,通过抑制IRF3途径而发挥抗炎效应,从而抑制LPS所诱导的KCs活化。
[Abstract]:Aim: to investigate the effect of LXR 伪 agonist T0901317 on the expression of IRF3 and GRIP1pLXR 伪 in kupffer cells stimulated by LPS, and to explore the mechanism of LXR 伪 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 伪 ligand agonist T09013175ugp / ml group was treated with LPS and T0901317. The expression of LXR 伪 GRIP1 and IRF3 protein in KCs were detected by Western bolt assay. The content of IFN 尾 -TNF- 伪 and IL-1 尾 in supernatant of KCs culture was determined by Elisa. Results the protein expression level of 1: LXR 伪 was the lowest in T0901317 treatment group (0.654 卤0.004) and LPS treatment group (0.008 卤0.003). The expression of LXR 伪 in the combined treatment group was significantly lower than that in the LXR 伪 agonist group, but it was significantly higher than that in the other two groups (0.977 卤0.019, 0.761 卤0.005, 0.908 卤0.011, 0.710 卤0.003, respectively) in the LPS group, and the highest in the LPS group (0.977 卤0.019, 0.761 卤0.005, respectively). The expression of LXR 伪 in the combined treatment group was significantly lower than that in the other two groups (0.908 卤0.011, 0.710 卤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 卤0.064 卤0.492 卤0.023) and LXR 伪 agonist group (0.468 卤0.019 卤0.425 卤0.002, respectively). Compared with the control group (129.6 卤17) and the LXR 伪 agonist group (112.8 卤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 伪 agonist group in the combined treatment group was 15.45 卤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 卤0.25 and 6.62 卤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- 伪 in the control group (5.05 卤0.17) and the combined treatment group (6.62 卤0.38). Conclusion: prophylactic application of LXR 伪 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.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R362

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