1.The effect of hydroquinone on autophagy of L-02 hepatocytes
Zuobing Zheng ; Wei Zhang ; Liang Xiong ; Tingting Liu ; Yangna Wu ; Jiahui Yang ; Gonghua Hu
Acta Universitatis Medicinalis Anhui 2022;57(9):1425-1430
Objective :
To study the effect of hydroquinone on autophagy of human L-02 hepatocytes.
Methods :
L-02 cells were treated with different concentrations of hydroquinone(0,10,20,40 and 80 μmol/L) for 24 hours, the formation of autophagosomes was observed by transmission electron microscope;autophagosome formation was traced by mCheery-GFP-LC3B fusion protein. Western blot experiment was used to detect the expression of autophagy marker proteins LC3,Beclin-1,P62;immunofluorescence experiment was used to detect the subcellular localization and expression of autophagy proteins LC3,Beclin-1,P62.
Results :
Under the transmission electron microscope,it was observed that the autophagosomes and lysosomes of L-02 cells increased after the action of hydroquinone. Under the fluorescence microscope,the number of GFP-LC3B punctate yellow spots increased with the increase of hydroquinone dose,while the red flake spots decreased. Western blot results showed that hydroquinone increased the ratio of LC3Ⅱ/LC3Ⅰ protein in L-02 cells,and the difference between treatment groups and the control group was statistically significant(P<0. 05). Hydroquinone had no significant effect on Beclin-1 and P62 protein expression in L-02 cells(P>0. 05). The results of immunofluorescence experiments showed that LC3 and Beclin-1 were both expressed in the nucleus and cytoplasm of L-02 cells,hydroquinone had no effect on the subcellular localization of LC3 and Beclin-1 in L-02 cells,while 40 and 80 μmol/L hydroquinone could induce the translocation of P62 in L-02 cells from the cytoplasm to the nucleus.
Conclusion
Hydroquinone induces the increase of autophagosomes in L-02 cells,which may be related to the obstacles to autophagosome clearance.