BET抑制剂(+)-JQ1通过调节NF-KB信号通路保护小鼠急性肝衰竭实验研究*

doi:10.3969/j.issn.1672-5069.2020.06.006

摘要/Abstract

摘要： 目的探讨溴结构域和额外末端结构域(BET)抑制剂(+)-JQ1对脂多糖/D-氨基半乳糖(LPS/D-Gal)诱导的小鼠急性肝衰竭(ALF)的保护作用及其作用机制。方法将47只C57BL/6小鼠随机分为对照组(n=15)、模型组(n=17)和(+)-JQ1干预组(n=15),采用腹腔注射LPS/D-Gal联合诱导ALF模型,其中在干预组造模前2 h腹腔注射(+)-JQ1。在造模4 h后每组处死5只小鼠,其余小鼠继续喂养至72 h,观察存活率。采用ELISA法检测血清TNF-α,采用RT-qPCR法和Western bloting法分别检测mRNA和蛋白表达。结果模型组72 h小鼠存活率为16.7%(2/12),(+)-JQ1干预组为60.0%(6/10,P>0.05);ALF组血清ALT水平为(2779.0±200.6)U/L,显著高于对照组【(44.9±2.5)U/L,P＜0.05】,血清AST水平为(2885.0±143.6)U/L,显著高于对照组【76.0±5.7) U/L,P＜0.05】,而(+)-JQ1干预组血清ALT和AST水平较模型组显著降低【分别为(948.7±46.3)U/L和(1704.0±42.1)U/L,P＜0.05】;ALF组肝组织TNF-αmRNA水平为(103.7±23.0),显著高于对照组【(1.2±0.2),P＜0.05】,IL-6 mRNA水平为(73.4±15.8) ,显著高于对照组【(1.1±0.1),P＜0.05】,IL-1B mRNA水平为(13.5±2.7) ,显著高于对照组【(1.0±0.1),P＜0.05】,而经(+)-JQ1干预后,三种细胞因子mRNA水平均较模型组显著降低【分别为(12.8±1.2)、 ( 11.7±0.7) 和 (1.5±0.3),P＜0.05】;ALF组血清TNF-α水平(631.6±57.5)U/L,显著高于对照组【(2.5±0.5)U/L,P＜0.05】,而(+)-JQ1干预组血清TNF-α水平为(139.8±8.1)U/L,显著低于ALF组(P＜0.05);ALF组肝组织P65和IKB蛋白表达显著增强,而(+)-JQ1干预后显著减弱。结论 (+)-JQ1对ALI小鼠肝脏具有保护作用,可能通过调节NF-KB信号通路下调促炎细胞因子的表达,从而减轻肝组织炎症反应。

关键词: 急性肝损伤, (+)-JQ1, 细胞因子, NF-KB信号通路, 小鼠

Abstract: Objective The aim of this experiment was to investigate the protective effect of bromo- and extra-terminal domain (BET) inhibitor, (+)-JQ1 on lipopolysaccharide / D-galactose (LPS / D-Gal) -induced acute liver failure (ALF) in mice and its mechanism. Methods 47 C57BL / 6 mice were randomly divided into control ((n=17), ALF model (n=15) and (+)-JQ1 intervention group (n=15). The ALF model were established by intraperitoneal injection of LPS / D-Gal, and mice in (+)-JQ1 intervention group were injected intraperitoneally with (+)-JQ1 2 hours before the induction of ALF. Four hours after LPS/D-Gal injection , 5 mice were sacrificed in each group and the remaining mice were fed for 72 hours to observe 72-hour survival rate. Serum TNF-α was detected by ELISA, and hepatic protein and gene mRNA were assayed by RT-qPCR and Western bloting, respectively. Results The 72-hour survival rate in model group was 16.7%(2/12), not significantly different compared to 60.0% (6/10,P>0.05) in (+)-JQ1-intervened group; serum ALT level in mice with ALF was (2779.0±200.6)U/L and serum AST level was (2885.0±143.6)U/L, both significantly higher than 【(44.9±2.5)U/L and (76.0±5.7) U/L,P＜0.05】 in control, while they decreased significantly to 【(948.7±46.3)U/L and (1704.0±42.1)U/L, respectively, P＜0.05】 in (+)-JQ1-intervened group; the hepatic tissue TNF-αmRNA was (103.7±23.0), significantly higher than 【(1.2±0.2),P＜0.05】, the IL-6 mRNA was (73.4±15.8) , much higher than 【(1.1±0.1), P＜0.05】, and the IL-1B mRNA was (13.5±2.7) , significantly higher than 【(1.0±0.1), P＜0.05】 in the control, while they decreased greatly to 【(12.8±1.2), ( 11.7±0.7) and (1.5±0.3), respectively, P＜0.05】 in (+)-JQ1-intervened group; serum TNF-α level was (631.6±57.5)U/L, much higher than 【(2.5±0.5)U/L, P＜0.05】 in the control, while it decreased to 【(139.8±8.1)U/L, P＜0.05】 in (+)-JQ1-intervened group; the expression of P65 and IKB in hepatic tissues in model intensified obviously, while they weaken greatly in (+)-JQ1-intervened group. Conclusion The bromo- and extra-terminal domain (BET) inhibitor, (+)-JQ1 has a protective effect on the liver in mice with LPS / D-Gal-induced ALF, which might be related to the regulation of NF-KB signaling pathway to reduce the expression of proinflammatory cytokines.

Key words: Acute liver failure, (+)-JQ1, Proinflammatory cytokine, NF-Kβ signaling pathway, Mice

黄鹤鸣, 刘彦君, 付荣, 石催催, 范建高, 张元元, 罗成, 李光明. BET抑制剂(+)-JQ1通过调节NF-KB信号通路保护小鼠急性肝衰竭实验研究^*[J]. 实用肝脏病杂志, 2020, 23(6): 781-784.

Huang Heming, Liu Yanjun, Fu Rong, et al. Protective effect of bromo- and extra-terminal domain inhibitor (+), JQ1 on LPS / D-Gal-induced acute liver failure in mice[J]. Journal of Practical Hepatology, 2020, 23(6): 781-784.

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BET抑制剂(+)-JQ1通过调节NF-KB信号通路保护小鼠急性肝衰竭实验研究^*

Protective effect of bromo- and extra-terminal domain inhibitor (+), JQ1 on LPS / D-Gal-induced acute liver failure in mice

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