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Objective To explore the level of autophagy induced by oxygen glucose deprivation/reperfusion(OGD/R) injury in L02 cell.Methods L02 cells were cultured to establish the model of OGD/R injury and simulate clinical hepatic ischemia-reperfusion injury.The L02 cells were randomly divided into 5 groups : normal control group, oxygen-glucose deprivation 6 h/reperfusion 1,3,6,12 h group (OGD 6 h/R 1,3,6,12 h).Then observe the form changes of the L02 cells by optical microscope.The appreciation of the company's relative L02 cells was detected by MTT.The expression of autophagy related proteins such as Beclin-1, LC3 and p62 were evaluated by Western blot.Results Compared with the normal control group, the form damaged and the cells proliferation activity of L02 cells in the OGD/R group were gradually increased in a time-dependent manner.Compared with the normal control group, autophagy related proteins LC3 , Beclin-1 were increased at OGD 6 h/R 1 h.The expression of LC3 was gradually increased as the time went on and was increased gradually at OGD 6 h/R 6 h, reached a peak at OGD 6 h/R 12 h(P<0.01).The expression of Beclin-1 was gradually increased as the time went on and was increased gradually at OGD 6 h/R 6 h and OGD 6 h/R 12 h (P<0.01).The expression of p62 had no obvious change at OGD 6 h/R 1 h and OGD 6 h/R 3 h, began to increase sharply at OGD 6 h/R 6 h and reached a peak at OGD 6 h/R 12 h(P<0.01).Conclusion Our data suggests that oxygen-glucose deprivation/reperfusion may increase the level of autophagy and lead to autophagic cell death in L02 cell.
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Objective To evaluate the effect of propofol on autophagy during oxygen-glucose deprivation and restoration (OGD/R) in human liver cells.Methods Human hepatic HL-7702 cells at the logarithmic growth phase were seeded into culture plates and randomly divided into 3 groups (n =12 each) using a random number table:control group (group C),OGD/R group,and propofol + OGD/R group (group P+OGD/R).The cells were cultured in normal culture medium in group C.In OGD/R and P+OGD/R groups,the cells were subjected to O2-glucose deprivation for 6 h followed by restoration of O2-glucose supply for 12 h.Propofol with a final concentration of 50 mmol/L was added at 10 min before oxygen-glucose deprivation.The cell viability was detected by MTT assay.The expression of autophagy-related proteins such as microtubule-associated protein light chain 3 (LC3) and Beclin-1 was evaluated by Western blot.Immunofluorescence was used to determine the number and distribution of autophagosomes.Results Compared with group C,the cell viability was significantly decreased,the expression of LC3 and Beclin-1 was significantly up-regulated (P＜0.05),and the number of autophagosomes was significantly increased in OGD/R and P+OGD/R groups.Compared with group OGD/R,the cell viability was significantly increased,the expression of LC3 and Beclin-1 was significantly down-regulated (P＜0.05),and the number of autophagosomes was significantly decreased in group P+OGD/R.Conclusion The mechanism by which propofol reduces OGD/R injury is probably related to inhibition of autophagy in human liver cells.