Effect of ropivacaine on autophagy and autolysosomes in HeLa cells

Objective:To investigate the effects of ropivacaine on autophagy and autolysosomes in HeLa cells.Methods:The HeLa,HeLa-GFP-LC3 and HeLa-RFP-LAMP1 cell lines were divided into the blank control group,starvation group(starving for 0,4 and 12 hours) and ropivacaine group(10 mg/mL ropivacaine treating for 0,4 and 12 hours).The induction of autophagy,morphology of autolysosomes and numbers of lysosomes in HeLa,HeLa-GFP-LC3 and HeLa-RFP-LAMP1 cell lines treated with ropivacaine were detected using the microscopic imaging technique combined with a variety of biochemical means.Results:Compared with the blank control cells,the LC3 puncta and LC3Ⅱ protein level significantly increased in ropivacaine group(P<0.05).Compared with the blank control cells,many autolysosomes in ropivacaine group and starvation group were identified after 4 hours of treatment,and the autolysosomes size in ropivacaine group was bigger than that in starvation group.The number of lysosomes in starvation group began to reduce at 4 hours of treatment,and recovered at 12 hours of treatment,but which in ropivacaine group gradually decreased and could not recover in HeLa cells.Conclusions:Ropivacaine can induce the autophagy,and damage autolysosomes.