目的 探讨花生四烯酸（ARA）经细胞色素P450（CYPs）及环氧合酶2（COX-2）代谢途径在非酒精性脂肪肝（NAFLD）相关肝纤维化中的作用，并从肝星形细胞（HSC）活化的角度探讨其机制。 方法 采用雄性C57BL/6J小鼠为研究对象，高糖高脂饲养（HFD）诱导小鼠NAFLD相关肝纤维化模型。给予小鼠每天皮下注射COX-2/可溶性环氧化物水解酶（sEH）双抑制剂PTUPB（5 mg/kg）。12周后观察COX-2/sEH双抑制对小鼠NAFLD相关肝纤维化的影响。记录小鼠体重变化；检测小鼠葡萄糖耐受性；HE染色观察小鼠肝组织病理损伤；油红O染色观察小鼠肝组织脂质堆积；Masson染色及Western blot法检测肝组织胶原含量；Real-time PCR法检测小鼠肝组织基质金属蛋白酶抑制因子相关基因表达；Western blot法检测PTUPB对棕榈酸（PA）诱导的肝星形细胞JS1活化的影响。 结果 PTUPB可显著降低HFD小鼠的体重，提高小鼠葡萄糖耐受性，减轻肝组织病理损伤、脂质堆积和胶原沉积；PTUPB亦可降低HFD小鼠肝组织基质金属蛋白酶抑制因子Timp1及Timp2的基因表达；在细胞水平观察到PTUPB可降低PA诱导的JS1细胞的活化。 结论 抑制ARA的COX-2/sEH代谢可减轻HFD诱导的小鼠NAFLD相关肝纤维化，其机制与抑制HSC的活化有关。
Objective To investigate the role of arachidonic acid (ARA) in liver fibrosis associated with non-alcoholic fatty liver disease (NAFLD) through cytochrome P450 (CYPs) and cyclooxygenase-2 (COX-2) metabolic pathways, and to explore the mechanism from the perspective of activation of hepatic astrocytes (HSC). Methods Male C57BL/6J mice were used as research subjects, and the mice model of NAFLD-related liver fibrosis was induced by high sugar and high-fat feeding (HFD). Mice were given a daily subcutaneous injection of COX-2/soluble epoxide hydrolase (sEH) dual inhibitor PTUPB (5 mg/kg). After 12 weeks, the effect of COX-2/sEH double inhibition on NAFLD-related liver fibrosis in mice was observed. The bodyweight of mice was recorded. Glucose tolerance of mice was detected. HE staining was used to observe pathological damage of liver tissue, and oil red O staining was used to observe lipid accumulation in liver tissue of mice. Masson staining and Western blot were used to detect collagen content in liver tissue. The expression of matrix metalloproteinase-inhibitor-related genes in mouse liver tissue was detected by Real- time PCR. Western blot was used to detect the effect of PTUPB on the activation of JS1 induced by palmitic acid (PA). Results PTUPB could significantly reduce the bodyweight of HFD mice, improve the glucose tolerance of mice, reduce the pathological damage of liver tissue, lipid accumulation, and collagen deposition. PTUPB could also decrease the expression of matrix metalloproteinase inhibitors Timp1 mRNA and Timp2 mRNA in liver tissues of HFD mice. PTUPB was observed to reduce PA-induced activation of JS1 cells at the cellular level. Conclusions Inhibition of ARA COX-2 /sEH metabolism can reduce HFD-induced NAFLD-related liver fibrosis in mice, and the mechanism is related to inhibition of HSC activation.
[1] Francque S, Szabo G, Abdelmalek MF, et al. Nonalcoholic steatohepatitis: the role of peroxisome proliferator-activated receptors [J]. Nat Rev Gastroenterol Hepatol, 2021, 18(1): 24-39. DOI: 10.1038/s41575-020-00366-5.
[2] Abdelmalek MF. Nonalcoholic fatty liver disease: another leap forward [J]. Nat Rev Gastroenterol Hepatol, 2021, 18(2): 85-86. DOI: 10.1038/s41575-020-00406-0.
[3] Funk CD. Prostaglandins and leukotrienes: advances in eicosanoid biology [J]. Science, 2001, 294(5548): 1871-1875. DOI: 10.1126/science.294.5548.1871.
[4] Parola M, Pinzani M. Liver fibrosis: Pathophysiology, pathogenetic targets and clinical issues [J]. Mol Aspects Med, 2019, 65: 37-55. DOI:10.1016/j.mam.2018.09.002.
[5] Mederacke I, Hsu CC, Troeger JS, et al. Fate tracing reveals hepatic stellate cells as dominant contributors to liver fibrosis independent of its aetiology [J]. Nat Commun, 2013, 4: 2823. DOI: 10.1038/ncomms 3823.
[6] Wagner K, Inceoglu B, Hammock BD. Soluble epoxide hydrolase inhibition, epoxygenated fatty acids and nociception [J]. Prostaglandins Other Lipid Mediat, 2011, 96(1-4): 76-83. DOI: 10.1016/j.prostag landins.2011.08.001.
[7] Zhou Y, Liu T, Duan JX, et al. Soluble epoxide hydrolase inhibitor attenuates lipopolysaccharide-induced acute lung injury and improves survival in mice [J]. Shock, 2017, 47(5): 638-645. DOI: 10.1097/SHK.0000000000000767.
[8] Zhou Y, Yang J, Sun GY, et al. Soluble epoxide hydrolase inhibitor 1-trifluoromethoxyphenyl-3- (1-propionylpiperidin-4-yl) urea attenuates bleomycin-induced pulmonary fibrosis in mice [J]. Cell Tissue Res, 2016, 363(2): 399-409. DOI: 10.1007/s00441-015-2262-0.
[9] Zarriello S, Tuazon JP, Corey S, et al. Humble beginnings with big goals: small molecule soluble epoxide hydrolase inhibitors for treating CNS disorders [J]. Prog Neurobiol, 2019, 172: 23-39. DOI: 10.1016/j.pneurobio.2018.11.001.
[10]Narumiya S. Physiology and pathophysiology of prostanoid receptors [J]. Proc Jpn Acad Ser B Phys Biol Sci, 2007, 83(9-10): 296-319. DOI: 10.2183/pjab/83.296.
[11]高枫, 王亚萍. 帕瑞昔布可缓解梗阻大鼠肾的纤维化 [J]. 基础医学与临床, 2007, 27(10): 1170-1171.
[12]P JJ, Manju SL, Ethiraj KR, et al. Safer anti-inflammatory therapy through dual COX-2/5-LOX inhibitors: A structure-based approach [J]. Eur J Pharm Sci, 2018, 121: 356-381. DOI: 10.1016/j.ejps. 2018. 06.003.
[13]Hye Khan MA, Hwang SH, Sharma A, et al. A dual COX-2/sEH inhibitor improves the metabolic profile and reduces kidney injury in Zucker diabetic fatty rat [J]. Prostaglandins Other Lipid Mediat, 2016, 125: 40-47. DOI: 10.1016/j.prostaglandins.2016.07.003.
[14]Li J, Zhou Y, Wang H, et al. COX-2/sEH dual inhibitor PTUPB suppresses glioblastoma growth by targeting epidermal growth factor receptor and hyaluronan mediated motility receptor [J]. Oncotarget, 2017, 8(50): 87353-87363. DOI:10.18632/oncotarget.20928.
[15]Zhang CY, Duan JX, Yang HH, et al. COX-2/sEH dual inhibitor PTUPB alleviates bleomycin-induced pulmonary fibrosis in mice via inhibiting senescence [J]. FEBS J, 2019. DOI: 10.1111/febs.15105.
[16]张君, 张晨宇, 孙晨晨, et al. COX-2/sEH双抑制剂PTUPB通过抑制内质网应激改善小鼠非酒精性脂肪性肝病 [J]. 中国临床解剖学杂志, 2020, 38(5): 562-567+573. DOI: 10.13418/j.issn.1001-165x. 2020. 05.014.
[17]Sun CC, Zhang CY, Duan JX, et al. PTUPB ameliorates high-fat diet-induced non-alcoholic fatty liver disease via inhibiting NLRP3 inflammasome activation in mice [J]. Biochem Biophys Res Commun, 2020, 523(4): 1020-1026. DOI: 10.1016/j.bbrc.2019.12.131.
[18]Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis [J]. Adv Drug Deliv Rev, 2017, 121: 27-42. DOI: 10.1016/j.addr.2017.05.007.
[19]Trautwein C, Friedman SL, Schuppan D, et al. Hepatic fibrosis: Concept to treatment [J]. J Hepatol, 2015, 62(1 Suppl): S15-24. DOI: 10.1016/j.jhep.2015.02.039.
[20]Skibba M, Hye Khan MA, Kolb LL, et al. Epoxyeicosatrienoic acid analog decreases renal fibrosis by reducing epithelial-to-mesenchymal transition [J]. Front Pharmacol, 2017, 8: 406. DOI: 10.3389/fphar.2017.00406.
[21]Li X, Chu G, Zhu F, et al. Epoxyeicosatrienoic acid prevents maladaptive remodeling in pressure overload by targeting calcineurin/NFAT and Smad-7 [J]. Exp Cell Res, 2020, 386(1): 111716. DOI: 10.1016/j.yexcr.2019.111716.
[22]Kuo YM, Hsu PC, Hung CC, et al. Soluble epoxide hydrolase inhibition attenuates excitotoxicity involving 14,15-epoxyeicosatrienoic acid-mediated astrocytic survival and plasticity to preserve glutamate homeostasis [J]. Mol Neurobiol, 2019, 56(12): 8451-8474. DOI: 10.1007/s12035-019-01669-8.
[23]唐保东, 徐雅, 刘思纯, 等. 选择性COX-2抑制剂塞来昔布对肝星状细胞增殖及活化的影响 [J]. 胃肠病学和肝病学杂志, 2007,16(5): 468-470.
[24]Chen L, Ji X, Wang M, et al. Involvement of TLR4 signaling regulated-COX2/PGE2 axis in liver fibrosis induced by Schistosoma japonicum infection [J]. Parasit Vectors, 2021, 14(1): 279. DOI: 10.1186/s13071-021-04790-7
[25]Castro RE, Diehl AM. Towards a definite mouse model of NAFLD [J]. J Hepatol, 2018, 69(2): 272-274. DOI: 10.1016/j.jhep.2018.05.002
[26]张丹, 吕俊衍, 邓溢, 等. 三种不同饮食干预建立非酒精性脂肪肝模型 [J]. 昆明医科大学学报, 2020, 41(1): 11-17. DOI: CNKI:SUN:KMYX.0.2020-01-003
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