Effects of NOD8 on H2O2-induced apoptosis in human hepatocytes |
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| Authors: | ZHENG Yuan-yuan YANG Wen-xin ZHOU Han HU Chao-feng |
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| Institution: | Department of Pathophysiology, Laboratory of Pathophysiology, State Administration of Traditional Chinese Medicine of The People’s Republic of China, School of Medicine, Jinan University, Guangzhou 510632, China. |
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| Abstract: | AIM: To observe the effects of NOD8 on H2O2-induced apoptosis in human L02 hepatocytes. METHODS: pEGFP-C2 and pEGFP-NOD8 plasmids were transfected into L02 cells by JetPRIME, respectively. The apoptosis of these transfected cells was induced by H2O2. The cells were divided into pEGFP-C2 group, pEGFP-C2+H2O2 group and pEGFP-NOD8+H2O2 group. MTT assay was used to detect the cell viability. NOD8 protein expression was determined by Western blotting. The cell apoptosis was observed by Hoechst 33342 staining and apoptotic rate was evaluated by flow cytometry. The caspase-3 activity was analyzed by a colorimetric method. RESULTS: L02 cells were stimulated by H2O2 at concentrations of 0.2～2 mmol/L for 6 h, and H2O2 at concentration of 1 mmol/L was chosen to induce apoptosis determined by MTT assay. The protein expression of NOD8 significantly increased in the cells transfected with pEGFP-NOD8 plasmid. More cellular nucleus with strong blue fluorescence by Hoechst 33342 staining in pEGFP-C2+ H2O2 group were observed, indicating that apoptosis was increased, while the apoptosis in pEGFP-NOD8+H2O2 group significantly reduced. The apoptotic rate in pEGFP-C2+H2O2 group was obviously increased, whereas that in pEGFP-NOD8+H2O2 group was significantly decreased. The caspase-3 activity in pEGFP-C2+H2O2 group was remarkably increased. By contrast, the activity of caspase-3 was significantly reduced in pEGFP-NOD8+H2O2 group.CONCLUSION: NOD8 inhibits H2O2-induced apoptosis in L02 cells and the mechanism may be related to inhibition of caspase-3 activity. |
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| Keywords: | L02 cells H2O2 NOD8 Caspase-3 |
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