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400038 重庆,第三军医大学军事预防医学院营养与食品安全研究中心,重庆市营养与食品安全重点实验室,重庆市医学营养研究中心
[张乾勇,E-mail: zqianyong@sina.com Zhang Qianyong, E-mail: zqianyong@sina.com
收稿:2016-09-26,
修回:2016-10-20,
纸质出版:2017-06
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黄娟, 曾祥龙, 张乾勇, 等. 白藜芦醇通过上调脂肪变性HepG2细胞Sirt3表达改善线粒体功能[J]. 第三军医大学学报, 2017,39(6):536-540.
Juan Huang, Xianglong Zeng, Qianyong Zhang, et al. Resveratrol improves mitochondrial function
黄娟, 曾祥龙, 张乾勇, 等. 白藜芦醇通过上调脂肪变性HepG2细胞Sirt3表达改善线粒体功能[J]. 第三军医大学学报, 2017,39(6):536-540. DOI: 10.16016/j.1000-5404.201609164.
Juan Huang, Xianglong Zeng, Qianyong Zhang, et al. Resveratrol improves mitochondrial function
目的
2
探讨白藜芦醇 (resveratrol,RSV) 对脂肪变性肝细胞线粒体功能的影响,以及沉默信息调节因子3(sirtuin 3,Sirt3) 在其中的重要作用。
方法
2
用0.2 mmol/L棕榈酸 (palmitic acid
PA) 处理HepG2细胞24 h建立肝细胞脂肪变性模型,再用40 μmol/L RSV和/或50 μmol/L Sirt3抑制剂3-TYP处理24 h。实验分为对照组、PA组、PA+RSV组、PA+RSV+3-TYP组、PA+3-TYP组。用相关试剂盒检测甘油三酯 (TG) 含量、沉默调节蛋白3(sirtuin 3,Sirt3) 活性、ATP合酶活性、ATP生成量以及线粒体基础呼吸率,使用DCFH-DA荧光探针检测细胞活性氧 (reactive oxygen species,ROS) 含量,Western blot检测Sirt3蛋白表达。Sirt3 siRNA处理HepG2细胞后再用PA和/或RSV处理,分析ROS和甘油三酯含量。
结果
2
相对于PA组,PA+RSV组HepG2细胞的甘油三酯、ROS含量明显降低 (
P
<
0.05),ATP合酶活性、ATP生成量和基础呼吸率明显提高 (
P
<
0.05)。而3-TYP或Sirt3 siRNA处理后,RSV的上述作用被削弱 (
P
<
0.05)。
结论
2
RSV通过上调脂肪变性肝细胞Sirt3的表达与活性而改善其线粒体功能。
Objective
2
To explore the influence of resveratrol (RSV) on mitochondrial function in induced steatosis in hepatic cells and investigate the role of sirtuin 3 (Sirt3) in it.
Methods
2
HepG2 cells were pretreated with palmitic acid (PA
0.2 mmol/L) for 24 h to establish hepatic steatosis cell model. Then
these hepatic steatosis cells were treated with 40 μmol/L RSV and/or 50 μmol/L 3-TYP (Sirt3 inhibitor) for 24 h. Then the cells were divided into 5 groups: control group
PA group
PA+RSV group
PA+RSV+3-TYP group
and PA+3-TYP group
respectively. The levels of triglycerides (TG) and ATP
Sirt3 activity
ATP synthase activity and basal oxygen consumption rate were measured using relevant kits. Reactive oxygen species (ROS) content was detected by DCFH-DA fluorescence probe. The expression of Sirt3 was measured by Western blotting.
Results
2
Compared with the PA group
the levels of TG and ROS in the PA+RSV group were decreased significantly (
P
<
0.05). On the contrary
the ATP synthase activity
ATP level and basal oxygen consumption rate were increased significantly (
P
<
0.05). However
the effects of RSV were impaired with 3-TYP or Sirt3 siRNA treatment (
P
<
0.05).
Conclusion
2
RSV can improve mitochondrial function by up-regulating the expression and activity of Sirt3 in steatosis in hepatic cells.
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