王磊, 郑敏, 张蔓, 吴智君, 赵文锦, 程娟. 内质网应激在二甲基甲酰胺致小鼠肝脏损伤中的作用[J]. 环境与职业医学, 2018, 35(12): 1123-1128. DOI: 10.13213/j.cnki.jeom.2017.18558
引用本文: 王磊, 郑敏, 张蔓, 吴智君, 赵文锦, 程娟. 内质网应激在二甲基甲酰胺致小鼠肝脏损伤中的作用[J]. 环境与职业医学, 2018, 35(12): 1123-1128. DOI: 10.13213/j.cnki.jeom.2017.18558
WANG Lei, ZHENG Min, ZHANG Man, WU Zhi-jun, ZHAO Wen-jin, CHENG Juan. Role of endoplasmic reticulum stress in mice liver injury induced by N, N-dimethylformamide[J]. Journal of Environmental and Occupational Medicine, 2018, 35(12): 1123-1128. DOI: 10.13213/j.cnki.jeom.2017.18558
Citation: WANG Lei, ZHENG Min, ZHANG Man, WU Zhi-jun, ZHAO Wen-jin, CHENG Juan. Role of endoplasmic reticulum stress in mice liver injury induced by N, N-dimethylformamide[J]. Journal of Environmental and Occupational Medicine, 2018, 35(12): 1123-1128. DOI: 10.13213/j.cnki.jeom.2017.18558

内质网应激在二甲基甲酰胺致小鼠肝脏损伤中的作用

Role of endoplasmic reticulum stress in mice liver injury induced by N, N-dimethylformamide

  • 摘要: 目的 研究内质网应激在二甲基甲酰胺(DMF)亚急性染毒致小鼠肝脏损伤中的作用及机制。

    方法 8周龄雄性C57BL/6小鼠20只,随机分为对照组、低剂量组、中剂量组、高剂量组,各5只。DMF灌胃剂量分别为150 mg/kg·bw(低)、450 mg/kg·bw(中)、1 350 mg/kg·bw(高),对照组给予同等体积生理盐水。连续灌胃28 d,1次/d。末次染毒结束后,次日解剖。生化分析仪检测小鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)的水平,HE染色观察肝脏组织病理学改变,Western blot法检测内质网应激标志性蛋白GRP94、GRP78及DMF代谢酶CYP2E1的表达水平。

    结果 DMF各染毒组小鼠体重增长受到抑制,与对照组相比差异均有统计学意义(P < 0.05),且随染毒剂量增加,抑制程度加重。与对照组相比,中、高剂量组小鼠肝脏系数增大,差异均有统计学意义(P < 0.05)。中、高剂量组血清ALT水平升高,与对照组相比,差异有统计学意义(P < 0.05),且高剂量组ALT水平较中剂量组高,差异有统计学意义(P < 0.05)。DMF染毒组AST水平也表现出升高,但低、中剂量组与对照组相比,差异均无统计学意义(P > 0.05),高剂量组与其他组相比,差异均有统计学意义(P < 0.05)。肝脏病理学检测显示,低剂量组小鼠肝小叶汇管区周围可见细胞水肿,细胞质疏松浅染,出现空泡;中剂量组出现大面积细胞水样变性,细胞肿胀,细胞核皱缩,肝窦扩张;高剂量组中央静脉周围出现大量细胞嗜酸性变,胞浆浓缩,细胞核固缩,出现嗜酸小体;对照组小鼠未见异常改变。Western blot结果显示,低、中、高剂量组的GRP94、CYP2E1表达水平升高,与对照组相比差异均有统计学意义(P < 0.05),且GRP94的表达水平随染毒剂量的升高呈剂量-效应关系。中、高剂量组GRP78表达水平与对照组相比,差异均有统计学意义(P < 0.05)。

    结论 DMF亚急性染毒可导致小鼠肝脏损伤,其机制可能与内质网应激有关。

     

    Abstract: Objective To study the role and mechanism of endoplasmic reticulum stress in subacute N, N-dimethylformamide (DMF) exposure induced liver injury in mice.

    Methods Twenty male C57BL/6 mice (8 weeks old) were randomly divided into control group (n=5), low-dose group (n=5), middle-dose group (n=5), and high-dose group (n=5). The mice in the exposure groups were administrated by gavage with DMF at 150 mg/kg·bw (low), 450 mg/kg·bw (middle), 1 350 mg/kg·bw (high), respectively, once a day for 28 days; the control group was administrated by gavage with normal saline at the same volume. All the animals were sacrificed the next day after last exposure. The levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured with automatic biochemistry analyzer. Histopathological changes of liver were observed after HE staining. Expression levels of endoplasmic reticulum stress markers (GRP94 and GRP78) and DMF metabolic enzyme (CYP2E1) were detected by Western blot.

    Results The body weight gain was significantly suppressed after repeated DMF administrations at various doses compared with the control group (P < 0.05), and the suppression was aggravated with the increase of exposure dose. Compared with the control group, the liver coefficients of mice in the middle-and high-dose groups were significantly increased (P < 0.05). The levels of serum ALT in the middle-and high-dose groups were significantly higher than that in the control group (P < 0.05), and the level of ALT in the high-dose group was higher than that in the middle-dose group (P < 0.05). The level of AST in the high-dose DMF exposure group was also increased compared with the control group (P < 0.05), but there were no significant differences between the low-or middle-dose groups and the control group (P > 0.05). The pathological examination results of liver showed hepatocellular edema and vacuolization in the low-dose group; a large area of vacuolar degeneration of cells, nucleus contraction, and sinus dilatation were observed in the middle-dose group; amounts of eosinophilic cells with cytoplasm concentration, nucleus contraction, and eosinophilic bodies appeared in centrilobular hepatocytes in the high-dose group; the mice in the control group did not show any abnormal change. The Western blot results showed that the expression levels of GRP94 and CYP2E1 in the low-, middle-, and high-dose groups were significantly higher than those in the control group (P < 0.05), and the expression level of GRP94 was increased in a dose-dependent manner. In addition, the expression levels of GRP78 in the middle-and high-dose groups were different from that in the control group (P < 0.05).

    Conclusion Subacute exposure to DMF could induce liver injury in mice, and the mechanism may be associated with endoplasmic reticulum stress.

     

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