抑制TRAF6基因表达对自身免疫性肝炎大鼠肝组织NF-κB信号通路相关蛋白表达的影响

doi:10.3969/j.issn.1672-5069.2021.04.004

摘要/Abstract

摘要： 目的探讨抑制TRAF6基因表达对自身免疫性肝炎(AIH)大鼠肝组织NF-κB信号通路相关蛋白表达的影响。方法随机将40只SD大鼠分为对照组、模型组、空载体组和TRAF6 shRNA干扰组,每组10只,采用特异性抗原S-100诱导建立大鼠AIH模型,分别给予生理盐水、转染空载体和转染TRAF6 shRNA处理。采用免疫组化法检测肝组织TRAF6蛋白表达,采用qRT-PCR法检测肝组织TRAF6 mRNA水平,采用ELISA法检测肝组织匀浆白介素1β(IL-1β)和白介素6(IL-6)水平,采用Western blot检测肝组织NF-κB信号通路相关基因蛋白表达。结果模型组大鼠肝组织TRAF6 mRNA水平为(6.2±0.4),显著高于正常组(1.0±0.2,P＜0.05),而TRAF6 shRNA干预组大鼠肝组织TRAF6 mRNA水平为(1.5±0.2),较模型组显著降低(P＜0.05);模型组大鼠肝组织匀浆IL-1β水平为(60.3±8.1)pg/mL,IL-6水平为(41.5±5.9)pg/mL,均显著高于正常组[(8.9±0.6)pg/mL和(15.6±0.6)pg/mL, P＜0.05];模型组大鼠肝组织IκBα、NF-κB p65、NF-κB p50、p-NF-κB p65和p-NF-κB p50蛋白表达相对水平分别为(1.3±0.1)、(1.6±0.1)、(1.3±0.1)、(1.3±0.1)和(1.1±0.1),均显著高于正常组[分别为(0.3±0.03)、(0.3±0.02)、(0.3±0.03)、(0.3±0.03)和(0.3±0.03),P＜0.05],而抑制了TRAF6表达组大鼠肝组织IL-1β为(31.5±4.0)pg/mL,IL-6为(27.4±4.2)pg/mL,IκBα、NF-κB p65、NF-κB p50、p-NF-κB p65和p-NF-κB p50蛋白表达分别为(1.0±0.1)、(0.9±0.05)、(0.7±0.1)、(0.8±0.1)和(0.5±0.05),均显著低于模型组(均P＜0.05)。结论抑制TRAF6基因表达可降低AIH大鼠肝内炎症水平,从而能改善肝组织损伤,其机制可能与调节了NF-κB信号通路的活化有关。

关键词: 自身免疫性肝炎, 肿瘤坏死因子受体相关因子6, NF-κB信号通路, 大鼠

Abstract: Objective The aim of this study was to investigate the impact of inhibition of tumor necrosis factor receptor associated factor 6 (TRAF6) gene expression on hepatic nuclear factor kappa-B (NF-κB)-related protein expression in rats with autoimmune hepatitis (AIH). Methods 40 rats were randomly divided into four groups, with 10 in each. The model of AIH was established in SD rats by specific antigen S-100, and the model rats were transfected with empty vector or TRAF6 shRNA vector. The expression of TRAF6-related proteins were detected by immunohistochemistry. The level of TRAF6 mRNA was detected by qRT-PCR. The IL-1β and IL-6 levels in liver homogenates were detected by ELISA. The expression of NF-κB signaling pathway related proteins in liver tissues was detected by Western blot. Results The hepatic TRAF6 mRNA level in model was (6.2±0.4), significantly higher than (1.0±0.2, P＜0.05) in control, while that in TRAF6 shRNA-intervened group decreased greatly as compared to in the model [(1.5±0.2), P＜0.05]; the liver homogenate IL-1βand IL-6 levels in the model were (60.3±8.1)pg/mL and (41.5±5.9)pg/mL, both significantly higher than [(8.9±0.6)pg/mL and (15.6±0.6)pg/mL, respectively, P＜0.05] in the control; the relative hepatic expression of IκBα, NF-κB p65, NF-κB p50, p-NF-κB p65 and p-NF-κB p50 in the model were (1.3±0.1), (1.6±0.1), (1.3±0.1), (1.3±0.1) and (1.1±0.1), all significantly higher than [(0.3±0.03), (0.3±0.02), (0.3±0.03), (0.3±0.03) and (0.3±0.03), P＜0.05] in the control, while they all significantly decreased in TRAF6-intervened group [(1.0±0.1),(0.9±0.05),(0.7±0.1),(0.8±0.1) and (0.5±0.05), P＜0.05] as compared to those in the model. Conclusion Inhibition TRAF6 gene expression could down-regulate the NF-κB signaling pathway, which might reduce liver inflammation of rats with AIH.

Key words: Autoimmune hepatitis, tumor necrosis factor receptor associated factor 6, Nuclear factor kappa-B signaling pathway, Rats

刘娜, 张华敏, 邓巧娟. 抑制TRAF6基因表达对自身免疫性肝炎大鼠肝组织NF-κB信号通路相关蛋白表达的影响[J]. 实用肝脏病杂志, 2021, 24(4): 468-471.

Liu Na, Zhang Huamin, Deng Qiaojuan. Impact of inhibition of TRAF6 gene expression on hepatic NF-κB-related protein expression in rats with autoimmune hepatitis[J]. Journal of Practical Hepatology, 2021, 24(4): 468-471.

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