外源性硫化氢对小鼠原代肝细胞内脂质自噬分解的影响

发布时间：2018-04-11 09:07

  本文选题：硫化氢 + 脂解　； 参考：《中国病理生理杂志》2017年10期

【摘要】：目的:研究硫化氢(H2S)供体GYY4137释放的外源性H2S对小鼠原代肝细胞内脂质自噬分解的影响。方法:采用2步原位灌流法分离C57BL/6小鼠原代肝细胞并分为4组:用正常培养基培养正常组细胞;模型组用含1.2 mmol/L油酸(溶于10%的BSA)的培养基培养48 h;H2S组和炔丙基甘氨酸(PAG)组则用1.2 mmol/L油酸(溶于10%的BSA)的培养液培养48 h,然后换无血清无酚红的RPMI-1640培养液(同时分别给予1 mmol/L GYY4137和200μmol/L PAG)处理6 h。收集各组细胞做LC3免疫荧光染色,拍摄荧光和相差图片;Western blot检测肝细胞中LC3-Ⅰ/Ⅱ的蛋白表达量;透射电镜观察肝细胞超微结构。结果:和模型组相比,H2S组肝细胞内LC3荧光颗粒和蛋白表达量增加,自噬溶酶体数量增加,细胞内空泡增多。结论:外源性H2S可促进脂肪变性肝细胞内脂质自噬分解。
[Abstract]:Aim: to study the effect of exogenous H _ 2S released by hydrogen sulfide (H _ 2S) donor GYY4137 on lipid autophagy in primary hepatocytes of mice.Methods: the primary hepatocytes of C57BL/6 mice were isolated by two-step in situ perfusion and divided into 4 groups: normal cells were cultured in normal culture medium;The model group was cultured with 1.2 mmol/L oleic acid (10% BSA-soluble medium) for 48 h in H2S group and propargyl glycine group (PAG) in 1.2 mmol/L oleic acid (10% BSA-soluble medium) for 48 h, and then replaced with serum-free RPMI-1640 medium (at the same time).They were treated with 1 mmol/L GYY4137 and 200 渭 mol/L PAG for 6 h, respectively.The expression of LC3- 鈪，

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