Regulatory effect of selenium on autophagy in preeclampsia trophoblast cells through the PI3K/Akt/mTOR signaling pathway

LIU Wan'er; ZHANG Lele; WANG Ting; GUO Wei

doi:10.3969/j.issn.1002-266X.2024.34.008

Abstract：Objective To explore the effects of selenium on autophagy and proliferation and migration of human tro-phoblast cells(HTR-8-Svneo)in preeclampsia(PE)through the PI3K/Akt/mTOR signaling pathway and the underlying mechanisms.Methods HTR-8-Svneo cells were cultured in vitro and randomly divided into the normal control group,hypoxia group,and hypoxia plus selenium group.HTR-8-Svneo cells in the control group were placed in a 37℃ CO2 incu-bator,HTR-8-Svneo cells in the hypoxia group were placed in a 37℃ hypoxia incubator,and HTR-8-Svneo cells in the hy-poxia plus selenium group were placed in a 37℃ hypoxia incubator with selenium supplementation(at the concentration of 10 µmol/L),and all groups were cultured for 24 h.CCK-8 assay kit was used to measure the viability of HTR-8-Svneo cells,the cell scratch test was used to detect the proliferation and migration abilities of HTR-8-Svneo cells,and Western blotting was used to detect the expression of PI3K/Akt/mTOR signaling pathway proteins(p-PI3K,p-Akt,p-mTOR)and autophagy-related proteins(LC3,Beclin-1,p62)in HTR-8-Svneo cells,and immunofluorescence was used to observe the expression of LC3 in HTR-8-Svneo cells.Results The OD values of cell viability in the control,hypoxia,and hypoxia plus selenium groups were 1.5±0.21,1.04±0.05,and 1.48±0.27,respectively;the cell viability was lower in the hy-poxia group than in the control group,and the cell viability was higher in the hypoxia plus selenium group than in the hy-poxia group(both P<0.05).The cell scratch healing rates in the control,hypoxia,and hypoxia plus selenium groups were 17.25%,6.08%,and 18.13%,respectively;the scratch healing rate in the hypoxia group was lower than that in the con-trol group,and the scratch healing rate in the hypoxia plus selenium group was higher than that in the hypoxia group(both P<0.05).There were statistically significant differences in the expression of pathway proteins and autophagy-related pro-teins in HTR-8-Svneo cells between the hypoxia group and the control group,and between the hypoxia plus selenium group and the hypoxia group(both P<0.05).The fluorescence expression levels of HTR-8-Svneo cells in the control,hypoxia,and hypoxia plus selenium groups were 73.26±4.18,134.83±3.03,and 74.7±12.37,respectively;the fluorescence expression level in the hypoxia group was higher than that in the control group,and the fluorescence expression level in the hypoxia plus selenium group was lower than that in the hypoxia group,with statistically significant differences(both P<0.05).Conclusion Selenium supplementation in the culture of HTR-8-Svneo cells can improve cell viability and prolif-eration and migration abilities,and the mechanism may be related to the regulation of autophagy in PE trophoblast cells through the PI3K/Akt/mTOR pathway.