Acta Anatomica Sinica

AAS ›› 2016, Vol. ›› Issue (2): 228-233.doi: 10.16098/j.issn.0529-1356.2016.02.013

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Inhibitory effect of low concentration ethanol on HepG2 cells

LIU Ying-jie 1* LI Jing-hua 2   

  1. 1. Department of Microbiology, Medical College of He’nan University, He’nan Kaifeng 475003，China; 2. Key Laboratory of Natural Medicine and ImmunoEngineering, He’nan University, He’nan Kaifeng 475003, China
  • Received:2015-09-17 Revised:2015-11-10 Online:2016-04-06 Published:2016-04-06
  • Contact: 刘英杰 E-mail:l-yingjie@henu.edu.cn

Abstract:

Objective To study the effect of low concentration ethanol on apoptosis of human hepatocellular carcinoma HepG2 cells, and to explore the mechanism. Methods  MTT assay was used to evaluate the cell inhibition rate. The cellular morphous was detected with AO/EB staining, and the ROS was detected with DCFH-DA staining, the mitochondrial membrane potential（ΔΨm）was detected with Rh123 staining,and the expression of Caspase-9 was detected by immunofluorescence technic. Apoptosis of HepG2 cells was quantified by flow cytometry using Annexin V/PI stain. Results Proliferation of HepG2 cells was inhibited by low concentration ethanol in a dosage and time dependent manner. Under HCS, some HepG2 cells underwent typical apoptotic morphologic change and the expression of ROS and Caspase-9 was increased. The mitochondrial membrane potential（ΔΨm）was decreased. Flow cytometry indicated 27.92% HepG2 cells were induced apoptosis and 5.4% HepG2 cells were necrotic after 6 hours incubation with 1.0mol/L ethanol, and 31.22% HepG2 cells were induced apoptosis and 15.1% HepG2 cells were necrotic after 6 hours incubation with 1.6mol/L ethanol. Conclusion The low concentration ethanol induces the apoptosis of HepG2 cells by the mitochondrial pathway. The mechanism is concerned with increasing ROS, decreasing the mitochondrial membrane potential and upgrading Caspase-9. 