亚硝酸钠增强自噬减轻肝脂肪变性的细胞脂肪蓄积

发布时间：2019-08-15 20:21

【摘要】：最近发现抑制自噬会加重肝细胞内脂肪蓄积,亚硝酸盐可以降低高脂饲养小鼠体内甘油三酯含量。因此,本实验目的在于观察亚硝酸盐对单纯性肝细胞脂肪变性的影响和自噬在其中的可能作用。首先用1.2 mmol·L-1油酸诱导人肝细胞系L-02细胞24 h建立肝细胞脂肪变性模型,再用0.2 mmol·L-1亚硝酸钠加或不加三甲基腺嘌呤(3-MA)或氯喹处理24 h。肝细胞脂质蓄积评价采用油红O染色和细胞内甘油三酯含量测定。1.2 mmol·L-1油酸诱导人肝细胞系L-02细胞24 h可以导致肝细胞明显的脂肪变性。用0.2 mmol·L-1亚硝酸钠处理L-02肝脂肪变性的细胞,可以明显减少肝细胞脂质蓄积,而3-MA可以减弱亚硝酸钠的这种效应;LC3-II免疫荧光、Western blot结果显示,亚硝酸钠促进L-02脂肪变性细胞LC3-II荧光颗粒和蛋白表达增加,氯喹同样可以增加这种现象;相差显微镜下显示细胞内空泡增多,溶酶体示踪剂荧光显微镜下发现,细胞溶酶体数量增多;电镜显示自噬溶酶体数量增加,氯喹可以减弱这种现象。这些结果表明,亚硝酸钠能够促进肝脂肪变性的L-02细胞自噬,促进脂质分解,减轻肝脂肪变性细胞脂质蓄积。
[Abstract]:Recently, it has been found that inhibition of autophagy can aggravate the accumulation of fat in hepatocytes, and nitrate can reduce the content of TG in mice fed with high fat. Therefore, the purpose of this study was to observe the effect of nitrate on simple hepatic steatosis and the possible role of autophagy in it. The hepatocytic steatosis model was established by using 1.2 mmol 路L-1 oleic acid for 24 h, and then treated with 0.2 mmol 路L-1 sodium nitroso with or without trimethyladenine (3-MA) or chloroquine for 24 h, and then treated with 0.2 mmol 路L-1 sodium nitroso or without trimethyladenine (3-MA) or chloroquine for 24 h. The lipid accumulation of hepatocytes was evaluated by oil red O staining and intracellular TG content. 1.2 mmol 路L-1 oleic acid induced obvious steatosis of human liver cell line L 鈮，

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