SEC62 通过介导自噬在对乙酰氨基酚诱导的小鼠肝损伤模型中的作用

摘要/Abstract

摘要： 目的探讨 SEC62 在对乙酰氨基酚(APAP)诱导的肝损伤小鼠模型中的作用机制。方法将24只 SPF 级 C57BL/6 品系的小鼠随机分为空白对照组(n=8)、 SEC62-Hep^ko APAP 组(n=8)及SEC62-Hep^flpox APAP 组(n=8)。SEC62-Hep^ko及SEC62-Hep^flpox小鼠腹腔注射APAP,模拟APAP诱导肝损伤进程(AILI)。空白对照组小鼠腹腔注射等量0.9% NaCl溶液,24 h后将上述各组小鼠处死,留取肝组织样本,进行 HE、免疫组织化学染色及肝组织流式细胞检测。结果 SEC62-Hep^flpox APAP 组小鼠肝脏病理显示多个大小不等的炎症坏死灶;而 SEC62-Hep^ko APAP 组小鼠肝组织中炎症坏死灶的数量及面积减少。免疫组化油红染色显示,SEC62-Hep^flpox APAP 组小鼠肝脏油红染色程度升高;而 SEC62-Hep^ko APAP 组小鼠肝组织油红染色程度减少。蛋白免疫印迹检测显示,SEC62-Hep^flpox APAP 组小鼠肝脏中自噬相关的蛋白 P62 的表达水平高于对照组(P=0.0037);与线粒体裂解相关的蛋白DRP-1及FIS-1的表述水平和与炎症反应相关的蛋白IL-1β的表达水平显著升高(均P<0.05)。与SEC62-Hep^flpox APAP 组相比,SEC62-Hep^ko APAP 组小鼠肝组织中自噬相关的蛋白 P62 的表达水平显著降低、与线粒体裂解相关的蛋白 DRP-1 及 FIS-1 的表述水平显著降低、与炎症反应相关的蛋白 IL-1β 的表达水平显著降低(均P<0.05)。流式细胞术检测显示,与空白对照组相比,在 SEC62-Hep^flpox APAP 组中,小鼠肝脏中 ROS 的表达水平显著升高(P<0.001)。而在 SEC62-Hep^ko APAP 组小鼠肝组织中,相比SEC62-Hep^flpox APAP 组,小鼠肝脏中 ROS 的表达水平显著降低(P<0.001)。结论 SEC62 通过介导自噬水平影响肝脏线粒体稳态及炎症反应,肝细胞中敲低 SEC62 在APAP 诱导的小鼠肝损伤模型中起到改善和保护作用。

关键词: SEC62, 线粒体稳态, APAP, 炎症反应, 药物性肝损伤, 自噬

Abstract: Objective To investigate the mechanism of SEC62 in mice model of liver injury induced by Acetaminophen (APAP). Methods 24 mice of SPF grade C57BL/6 strain were randomly divided into blank control group (n=8), SEC62-Hepko APAP group (n=8) and SEC62-Hepflpox APAP group (n=8). Eight SEC62-Hepko and SEC62-Hepflpox mice of SPF C57BL/6 strain were injected with APAP to establish APAP Induced Liver Injury (AILI). Mice in the blank control group were injected with normal saline intraperitoneally, and the liver tissue was collected 24 hours later, and pathological, immunohistochemical staining and flow cytometry of liver tissue were performed. Results Compared with blank control group, liver pathology of mice in SEC62-Hepflpox APAP group exhibited multiple inflammatory and necrotic area of different sizes. In the SEC62-Hepko APAP group, the number and area of inflammatory necrotic lesion in liver tissue decreased. Immunohistochemical oil red staining showed that the degree of oil red staining in liver of mice in SEC62-Hepflpox APAP group was higher than that in blank control group. The degree of oil red staining in liver tissue of SEC62-Hepko APAP group decreased. Western blot analysis showed that compared with control group, the expression level of autophagy related protein P62 in liver of SEC62-Hepflpox APAP group was significantly increased (P=0.0037). The expression levels of the proteins DRP-1 and FIS-1 associated with mitochondrial cleavage and the expression level of IL-1β, a protein associated with inflammation, was significantly increased (all P<0.05). Compared with those in SEC62-Hepflpox APAP group, the expression level of autophagy related protein P62 in liver of mice in SEC62-Hepko APAP group was significantly decreased , the expression levels of the proteins DRP-1 and FIS-1 associated with mitochondrial cleavage were significantly decreased ,and the expression level of IL-1β, a protein associated with inflammation, was significantly decreased (all P<0.05). Flow cytometry showed that ROS level in liver of mice in SEC62-Hepflpox APAP group was significantly increased compared with blank control group (P<0.001). In the liver tissues of mice in SEC62-Hepko APAP group, the level of ROS in the liver of mice was significantly decreased compared with that in SEC62-Hepflpox APAP group (P<0.001). Conclusion SEC62 can affect mitochondrial homeostasis and inflammation in liver by mediating autophagy level. SEC62 knockout in hepatocytes can improve and protect the liver injury induced by APAP.

Key words: SEC62, Mitochondrial homeostasis, APAP, Inflammatory response, Drug-induced liver injury, Autophagy

林俊超, 刘俞彤, 周威, 蒋炜, 何杰. SEC62 通过介导自噬在对乙酰氨基酚诱导的小鼠肝损伤模型中的作用[J]. 肝脏, 2025, 30(8): 1151-1155.

LIN Jun-chao, LIU Yu-tong, ZHOU wei, JIANG Wei, HE Jie. SEC62 exacerbates hepatocyte inflammation through its involvement in autophagy regulation in a murine model of paracetamol-induced liver injury[J]. Chinese Hepatolgy, 2025, 30(8): 1151-1155.

/ 推荐

参考文献

[1] Patel S J, Milwid J M, King K R, et al. Gap junction inhibition prevents drug-induced liver toxicity and fulminant hepatic failure[J]. Nat Biotechnol, 2012, 30(2):179-183.
[2] Shen T, Liu Y, Shang J, et al. Incidence and etiology of drug-induced liver injury in Mainland China[J]. Gastroenterology, 2019, Jun156(8):2230-2241.e11.
[3] Weaver R J, Blomme E A, Chadwick A E, et al. Managing the challenge of drug-induced liver injury: a roadmap for the development and deployment of preclinical predictive models[J]. Nat Rev Drug Discov, 2020, 19(2):131-148.
[4] Hunt C M, Papay J I, Stanulovic V, et al. Drug rechallenge following drug-induced liver injury[J]. Hepatology, 2017, 66(2):646-654.
[5] Su S, Shi Y T, Chu Y, et al.Sec62 promotes gastric cancer metastasis through mediating UPR-induced autophagy activation. Cellular and molecular life sciences[J]. CMLS, 2022, 79:133.
[6] Jin Y, Han Y, Yang S, et al. Endoplasmic reticulum- resident protein Sec62 drives colorectal cancer metastasis via MAPK/ATF2/UCA1 axis[J]. Cellproliferation, 2022, 55:e13253.
[7] Linxweiler M, Schick B, Zimmermann R. Let′s talk about Secs: Sec61,Sec62 and Sec63 in signal transduction, oncology and personalized medicine[J]. Signal transduction and targeted therapy, 2017, 2:17002.
[8] Fumagalli F, Noack J, Bergmann T J, et al. Translocon component Sec62 acts in endoplasmic reticulum turnover during stress recovery[J]. Nature cell biology, 2016, 18:1173- 1184.
[9] Yan M Z, Huo Y Z, Yin S T, et al. Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions[J]. Redox Biol, 2018, 17:274-283.
[10] Dunn W, Shah V H.Pathogenesis of alcoholic liver disease[J]. Clin Liver Dis, 2016, 20:445-56.
[11] Leise M D, Poterucha J J, Talwalkar J A. Drug-induced liver injury[J]. Mayo Clin Proc, 2014, 89:95-106.
[12] Jaeschke H, Ramachandran A. Acetaminophen hepatotoxicity: paradigm for understanding mechanisms of drug-induced liver injury[J]. Annu Rev Pathol, 2024, 19:453-478.
[13] Marchi S, Guilbaud E, Tait S W G, et al. Mitochondrial control of inflammation[J]. Nat Rev Immunol, 2023, 23(3):159-173.
[14] Chao X, Wang S, Zhao K, et al. Impaired TFEB-mediated lysosome biogenesis and autophagy promote chronic ethanol-induced liver injury and steatosis in mice[J]. Gastroenterology, 2018, 155(3):865-879.e12.
[15] Peng Y, Shapiro S L, Banduseela V C, et al. Increased transport of acetyl-CoA into the endoplasmic reticulum causes a progeria-like phenotype[J]. Aging cell, 2018, 17:e12820.