RESUMO
AIM:To evaluate the effect of exogenous hydrogen sulfide ( H2 S) from GYY4137 on lipophagy in mouse primary hepatocytes .METHODS:The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion method were divided into 4 groups:the cells in control group were incubated with normal medium; the cells in model group were incubated with 1.2 mmol/L oleic acid (OA) for 48 h;the cells in H2S group or propargylglycine (PAG) group were incubated with 1.2 mmol/L OA for 48 h followed by serum-free phenol red-free RPMI-1640 medium which con-tained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h.The cells were collected to conduct immunofluorescence staining of LC3 and photography under fluorescence microscope , phase-contrast microscope or transmission electron microscope . The protein expression of LC 3-Ⅰ/Ⅱin the hepatocytes was determined by Western blot .RESULTS:In contrast with the model group, the fluorescent particles of LC3, the protein expression of LC3, the number of autophagic lysosome and vacuoles in hepatocytes in H 2 S group increased .CONCLUSION: In steatosis hepatocytes , exogenous H 2 S promotes the lipophagy .
RESUMO
AIM:To evaluate the effect of exogenous hydrogen sulfide ( H2 S) from GYY4137 on lipophagy in mouse primary hepatocytes .METHODS:The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion method were divided into 4 groups:the cells in control group were incubated with normal medium; the cells in model group were incubated with 1.2 mmol/L oleic acid (OA) for 48 h;the cells in H2S group or propargylglycine (PAG) group were incubated with 1.2 mmol/L OA for 48 h followed by serum-free phenol red-free RPMI-1640 medium which con-tained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h.The cells were collected to conduct immunofluorescence staining of LC3 and photography under fluorescence microscope , phase-contrast microscope or transmission electron microscope . The protein expression of LC 3-Ⅰ/Ⅱin the hepatocytes was determined by Western blot .RESULTS:In contrast with the model group, the fluorescent particles of LC3, the protein expression of LC3, the number of autophagic lysosome and vacuoles in hepatocytes in H 2 S group increased .CONCLUSION: In steatosis hepatocytes , exogenous H 2 S promotes the lipophagy .