糖原合酶激酶3β抑制剂TDZD-8对急性肝衰竭小鼠肝组织炎症的保护作用研究*

doi:10.3969/j.issn.1672-5069.2023.03.006

摘要/Abstract

摘要： 目的探索糖原合酶激酶3β(GSK3β)抑制剂TDZD-8对急性肝衰竭(ALF)小鼠的保护作用。方法将24只小鼠随机分为对照组、模型组和TDZD-8处理组,给予D-Gal和LPS腹腔注射,制备ALF模型,分别给予生理盐水或TDZD-8干预。采用ELISA法检测肝组织匀浆肿瘤坏死因子-α(TNF-α )和白介素-1β(IL-1β)水平,采用蛋白印迹法检测肝组织IL-18、IL-1β、UNC－51样激酶1(ULK1)、Beclin 1和P62蛋白表达。结果模型组血清ALT、AST和TBIL水平分别为(3460.8±105.2)U/L、(2710.4±84.3)U/L和(93.8±1.1)μmol/L,显著高于对照组【分别为(28.1±4.2)U/L、(25.78±2.5)U/L和(3.4±0.4)μmol/L,P<0.05】,而TDZD-8处理组各指标均显著降低【分别为(370.1±7.1)U/L、(277.3±20.3)U/L和(35.1±0.8)μmol/L,P<0.05】;模型组肝组织匀浆TNF-α、IL-1β和血清无细胞游离(cf)-DNA水平分别为(3004.6±239.2)pg/mL、(469.6±56.7)pg/mL和(772.2±192.2)ng/mL,经TDZD-8处理后,各指标均显著降低【分别为(2353.3±284.7)pg/mL、(339.2±48.7)pg/mL和(180.0±15.4)ng/mL,P<0.05】;模型组肝组织IL-18和IL-1β相对表达量较对照组显著增强(P<0.05),而TDZD-8处理组小鼠蛋白相对表达量较模型组显著减弱(P<0.05);模型组肝组织ULK1和Beclin1相对表达量均显著低于对照组(P<0.05),而P62蛋白表达显著高于对照组(P<0.05),经TDZD-8处理,肝组织ULK1和Beclin1相对表达量显著高于模型组,而P62蛋白表现显著降低(P<0.05)。结论在急性肝衰竭小鼠,抑制GSK3β活性能够上调自噬水平,改善肝组织炎症因子的释放,从而起到保护肝脏作用。

关键词: 急性肝衰竭, 自噬, 糖原合酶激酶3β, 细胞因子

Abstract: Objective The aim of this study was to investigate the protective effect of glycogen synthase kinase 3β (GSK3β) inhibitor TDZD-8 on liver injuries in mice with D-Gal/LPS-induced acute liver failure(ALF). Methods Twenty-four male mice were randomly divided into control, model and TDZD-8-intervened group, with eight in each. The model of ALF was established by intraperitoneal injection of D-Gal/LPS. Serum free DNA (cf-DNA) was detected by fluorescent dye, and liver homogenate tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were assayed by ELISA. The hepatic expression of L-18, IL-1β, UNC-51-like kinase 1 (ULK1), Beclin 1 and P62 were detected by Western blotting. Results Serum ALT, AST and total bilirubin levels in model group were (3460.8±105.2)U/L, (2710.4±84.3)U/L and (93.8±1.1)μmol/L, all significantly higher than [(28.1±4.2)U/L,(25.78±2.5)U/L and (3.4±0.4)μmol/L, respectively, P<0.05] in the control, while they all greatly decreased in TDZD-8-intervened group [(370.1±7.1)U/L, (277.3±20.3)U/L and (35.1±0.8)μmol/L, respectively, P<0.05]; the liver homogenate TNF-α, IL-1β and serum cf-DNA levels in the model were (3004.6±239.2)pg/mL, (469.6±56.7)pg/mL and (772.2±192.2) ng/mL, while they all greatly decreased in TDZD-8-intervened group [(2353.3±284.7)pg/mL, (339.2±48.7)pg/mL and (180.0±15.4)ng/mL, respectively, P<0.05]; the hepatic expression of IL-18 and IL-1β in the model intensified greatly as compared to in the control (P<0.05), while they both became weaker in TDZD-8-intervened group (P<0.05); the relative hepatic expression of ULK1 and Beclin1 obviously decreased(P<0.05), and the hepatic expression of P62 protein increased greatly(P<0.05)compared to in the control, while the ULK1 and Beclin1 expression greatly intensified, and the P62 expression became weaker (P<0.05) in TDZD-8-intervened mice. Conclusion The inhibition of GSK3β activity in a mouse model of acute liver failure could protect the liver injuries, which might be related to the up-regulation of autophagy and the inhibition of inflammatory cytokine release.

Key words: Acute liver failure, Glycogen synthase kinase3β, Autophagy, Cytokines

张丹眉, 石春霞, 陈倩, 张璐懿, 张清奇, 邹旭晨, 龚作炯. 糖原合酶激酶3β抑制剂TDZD-8对急性肝衰竭小鼠肝组织炎症的保护作用研究^*[J]. 实用肝脏病杂志, 2023, 26(3): 324-327.

Zhang Danmei , Shi Chunxia , Chen Qian, et al.. Protective effect of glycogen synthase kinase 3β inhibitor TDZD-8 on liver injuries in mice with D-Gal/LPS-induced acute liver failure[J]. Journal of Practical Hepatology, 2023, 26(3): 324-327.

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糖原合酶激酶3β抑制剂TDZD-8对急性肝衰竭小鼠肝组织炎症的保护作用研究^*

Protective effect of glycogen synthase kinase 3β inhibitor TDZD-8 on liver injuries in mice with D-Gal/LPS-induced acute liver failure

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