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

doi:10.13418/j.issn.1001-165x.2020.05.014

中国临床解剖学杂志 ›› 2020, Vol. 38 ›› Issue (5) : 562-567. DOI: 10.13418/j.issn.1001-165x.2020.05.014

张君¹，　张晨宇²，　孙晨晨²，　田清¹，杨慧慧²，刘昱镳²，杨金桐²，周勇²，刘绍坤³

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COX-2/sEH dual inhibitor PTUPB alleviates non-alcoholic fatty liver disease in mice by inhibiting endoplasmic reticulum stress

ZHANG Jun¹, ZHANG Chen-yu², SUN Chen-chen², TIAN Qing¹, YANG Hui-hui², LIU Yu-biao², YANG Jin-tong², ZHOU Yong², LIU Shao-kun³

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摘要

目的　探讨环氧合酶2（COX-2）和可溶性环氧化物水解酶（sEH）的双抑制剂PTUPB对非酒精性脂肪性肝病（NAFLD）小鼠肝脏的保护作用及机制。方法　将32只C57BL/6雄性小鼠随机分为对照组（Control）、双抑制剂对照组（PTUPB）、高糖高脂饲料组（HFD）和双抑制剂治疗组（HFD+PTUPB）。其中PTUPB组和HFD+PTUPB组每天皮下注射COX-2/sEH双抑制剂PTUPB（5 mg/kg），Control组和HFD组皮下注射PTUPB的溶剂PEG400。12周后，观察各组小鼠的体重变化；HE染色、油红O染色和Masson染色分别观察小鼠肝组织病理改变、脂质堆积及胶原沉积；Western blot法检测小鼠肝组织内质网应激相关蛋白IRE-1α和XBP-1s的表达；Real-time PCR检测棕榈酸（PA）处理的小鼠肝细胞（AML-12）内质网应激相关基因（Perk、Ire-1α、Xbp-1s和Aft-6）的表达。结果　与HFD组小鼠相比，HFD+PTUPB组小鼠体重及体重变化率显著降低，肝组织病理损伤、脂质堆积和胶原沉积明显改善，IRE-1α和XBP-1s的蛋白表达显著下调；进一步在细胞水平观察到，PTUPB预处理可降低PA诱导的AML-12细胞内质网相关基因的表达。结论　抑制COX-2和sEH可减轻HFD诱导的小鼠NAFLD，其机制与抑制肝细胞内质网应激有关。

Abstract

Objective　To investigate the protective role and mechanism of cyclooxygenase-2 (COX-2) and soluble epoxide hydrolase (sEH) dual inhibitor PTUPB in non-alcoholic fatty liver disease (NALFD) in mice. Methods Thirty-two male C57BL/6 mice were randomly divided into the following groups: a control group, a PTUPB group, a HFD group, and a HFD + PTUPB group. Mice in the PTUPB group and the HFD + PTUPB group were subcutaneously injected with COX-2/sEH dual inhibitor PTUPB (5 mg/kg/d). Mice in the control group and the HFD group were subcutaneously injected with PEG400. After 12 weeks, the weight change of mice was observed. HE, Oil red O, and Masson staining were used to observe liver tissue pathology morphologic changes, lipid accumulation, and collagen deposition, respectively. The protein expression of IRE-1α and XBP-1s in the liver of NAFLD mice was detected by Western blot. The expression of genes related to endoplasmic reticulum stress (Perk，Ire-1α，Xbp-1s and Aft-6) in AML-12 treated with palmitic acid (PA) was detected by Real-time PCR. Results Compared with the HFD group, the body weight and weight change rate of mice in the HFD + PTUPB group significantly reduced. The liver pathological damage, lipid accumulation, and collagen deposition significantly ameliorated, and the protein expressions of IRE-1α and XBP-1s significantly down-regulated in the HFD + PTUPB group. In vitro, PTUPB pretreatment could reduce the expression levels of genes related to endoplasmic reticulum stress in AML-12 cells PA-induced. Conclusions Dual inhibition of COX-2 and sEH attenuates HFD-induced NALFD by suppressing endoplasmic reticulum stress in mice.

导出引用

张君, 张晨宇, 孙晨晨, 田清, 杨慧慧, 刘昱镳, 杨金桐, 周勇, 刘绍坤. 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 https://doi.org/10.13418/j.issn.1001-165x.2020.05.014

ZHANG Jun, ZHANG Chen-yu, SUN Chen-chen, TIAN Qing, YANG Hui-hui, LIU Yu-biao, YANG Jin-tong, ZHOU Yong, LIU Shao-kun. COX-2/sEH dual inhibitor PTUPB alleviates non-alcoholic fatty liver disease in mice by inhibiting endoplasmic reticulum stress[J]. Chinese Journal of Clinical Anatomy. 2020, 38(5): 562-567 https://doi.org/10.13418/j.issn.1001-165x.2020.05.014

中图分类号： R333.4

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