二乙基二硫代氨基甲酸通过Insr/Akt通路改善代谢相关脂肪性肝炎体内和体外实验研究*

doi:10.3969/j.issn.1672-5069.2025.04.006

摘要/Abstract

摘要： 目的探讨二乙基二硫代氨基甲酸(DDC)通过调控Insr/Akt信号通路改善代谢相关脂肪性肝炎(MASH)的分子机制。方法将C57BL/6小鼠随机分为对照组、MASH组和DDC干预组,每组6只。采用胆碱缺乏、L-氨基酸替代蛋白(CDAA)饲料喂养建立MASH模型或在造模同时给予DDC灌胃干预。采用RT-PCR和蛋白免疫印迹法检测肝组织胰岛素受体(Insr)和促凋亡分子Bax表达。采用棕榈酸(PA)诱导建立小鼠AML12细胞脂毒性模型,同时给予DDC干预。采用蛋白免疫印迹法检测AML12细胞Insr、Akt、pAkt、Bcl2和Bax蛋白表达。结果 MASH组小鼠肝组织Insr基因水平为(0.38±0.13),较对照组显著降低[(1.03±0.27),P<0.05];MASH组小鼠肝组织Insr蛋白表达水平为(0.61±0.11),较对照组显著降低[(1.68±0.58),P<0.05],而DDC干预组小鼠肝组织Insr蛋白表达水平为(1.03±0.11),较MASH组显著升高(P<0.05);MASH组小鼠肝组织Bax蛋白表达水平为(1.14±0.39),较对照组显著增强[(0.47±0.26),P<0.05],而DDC干预组小鼠肝组织Bax蛋白表达水平为(0.66±0.19),较MASH组显著降低(P<0.05);PA组AML12细胞Insr蛋白表达水平为(0.38±0.13),较对照组显著降低[(0.74±0.21),P<0.05],而DDC干预组AML12细胞Insr蛋白表达水平为(0.71±0.20),较PA组显著升高(P<0.05);PA组AML12细胞pAkt与Akt蛋白表达水平比值为(0.28±0.07),较对照组显著降低[(0.69±0.06),P<0.05];25 μM或50 μM DDC干预组AML12细胞pAkt与Akt蛋白表达水平比值分别为(0.85±0.02)和(0.97±0.04),较PA组显著升高(P<0.05); PA组AML12细胞Bcl2与Bax蛋白表达水平比值为(1.28±0.29),较对照组显著降低[(1.74±0.10),P<0.05],而DDC干预组AML12细胞Bcl2与Bax蛋白表达水平比值为(2.30±0.78),比PA组显著升高(P<0.05)。结论 CDAA饮食诱导的MASH小鼠肝组织Insr/Akt通路被抑制,细胞凋亡增加。DDC可能通过上调MASH小鼠肝组织Insr/Akt信号通路,抑制肝细胞凋亡,进而改善MASH。

Abstract: Objective The aim of this experiment was to investigate molecular mechanism of diethyldithiocarbamate (DDC) in improving metabolic dysfunction associated steatohepatitis (MASH) by regulating Insr/Akt signaling pathway. Methods 18 male C57BL/6 mice were randomly divided into control, MASH and DDC-intervened group (n=6 in each) and MASH model was established by feeding a choline-deficient, L-amino acid-defined (CDAA) diet, and the intervention was carried out by DDC gavage simultaneously. The expression of insulin receptor (Insr) and pro-apoptotic molecule, e.g., Bax in mouse liver tissue were detected by real-time PCR and Western blot. Model of lipotoxicity was established by induction of palmitic acid (PA) on AML12 cells, and was treated with DDC simultaneously. The expression of Insr, Akt, pAkt, Bax and Bcl2 were detected by Western blot. Results Insr mRNA level in liver tissue in MASH group was (0.38±0.13), significantly lower than [(1.03±0.27), P<0.05] in the control; the expression of Insr protein in liver tissues in MASH group was (0.61±0.11), significantly lower than [(1.68±0.58),P<0.05] in the control, while the expression of Insr protein in liver tissues in DDC-intervened group was (1.03±0.11), significantly higher than that in MASH group (P<0.05); the expression of Bax protein in liver tissue in MASH group was (1.14±0.39), significantly higher than [(0.47±0.26), P<0.05] in the control; the expression of Bax protein in liver tissues in DDC-intervened group was (0.66±0.19), significantly lower than that in MASH group (P<0.05); the expression of Insr protein in AML12 cells in PA group was (0.38±0.13), significantly lower than [(0.74±0.21), P<0.05] in the control, while the expression of Insr protein in AML12 cells in DDC-intervened group was (0.71±0.20), significantly higher than that in PA group (P<0.05); the ratio of pAkt/Akt protein in AML12 cells in PA group was (0.28±0.07), significantly lower than [(0.69±0.06), P<0.05] in the control, while the ratio of pAkt/Akt protein in AML12 cells in 25 μM or in 50 μM DDC-intervened groups were (0.85±0.02) and (0.97±0.04), both significantly increased than in PA group (P<0.05); the ratio of Bcl2/Bax in AML12 cells in PA group was (1.28±0.29), much lower than [(1.74±0.10),P<0.05] in the control, while the ratio of Bcl2/Bax in AML12 cells in DDC-intervened group was (2.30±0.78), much higher than that in PA group (P<0.05). Conclusion The Insr/Akt pathway is inhibited and apoptosis is increased in liver tissues in mice with CDAA diet-induced MASH, and DDC might inhibit hepatocyte apoptosis by stimulating Insr/Akt signaling pathway, thereby improving MASH progression.

Key words: Metabolic dysfunction associated steatohepatitis, Diethyldithiocarbamate, Insulin receptor, AML12 cells, Mices

齐一菲, 于庆红, 白世锦, 刘琳, 范旭, 王萍, 丛敏, 刘天会. 二乙基二硫代氨基甲酸通过Insr/Akt通路改善代谢相关脂肪性肝炎体内和体外实验研究^*[J]. 实用肝脏病杂志, 2025, 28(4): 501-504.

Qi Yifei, Yu Qinghong, Bai Shijin, et al. Diethyldithiocarbamate improves metabolic dysfunction associated steatohepatitis through regulating Insr/Akt pathway in mice and in vitro[J]. Journal of Practical Hepatology, 2025, 28(4): 501-504.

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二乙基二硫代氨基甲酸通过Insr/Akt通路改善代谢相关脂肪性肝炎体内和体外实验研究^*

Diethyldithiocarbamate improves metabolic dysfunction associated steatohepatitis through regulating Insr/Akt pathway in mice and in vitro

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