Effects Of PI3K/Akt Signaling Pathway On Acetaldehyde-stimulated Proliferation Of Hepatic Stellate Cells HSC-t6

Objective:The rat HSC-T6 was stimulated with acetaldehyde,To explore the possible relationship between PI3 K /Akt signaling pathway and the proliferation of rat HSC-T6.Methods:(1)The rat HSC-T6 were randomly divided into six groups:blank control group(group A): Used DMEM culture solution with 10% fetal bovine serum(FBS)to culture;The acetaldehyde group(group B):Added acetaldehyde on the basis of group A and made the final concentration be200μmol/L;experimental group(C、D、E、F):Added with different concentrations of PI3K/Akt pathway specific inhibitor LY294002(25、50、75、100 μmol/L)on the basis of group B,Pretreatment for 1h,then add200μmol/L acetaldehyde(final concentration),the intervention time of 24hours(add every 12hours).CCK-8was applied to detect the proliferation activity of rat HSC-T6.(2)The rat HSC-T6 were randomly divided into three groups:the blank control group(added with DMEM culture solution with 10% fetal bovine serum to culture);acetaldehyde group(add with acetaldehyde on the basis of blank control group and made the final concentration be200μmol/L);acetaldehyde+LY294002 group(add 50μmol/L PI3K/Akt pathway specific inhibitor LY294002 on the basis of acetaldehyde group).Western-blotting was used to detect the expression of PI3 K and p-Akt protein in rat HSC-T6.Results:(1)Rat HSC-T6 cells was observed under inverted microscope:Blank control group(group A)cell growth was relatively uniform,not easy to grow into a group;After the addition of acetaldehyde,the acetaldehyde group(group B)cell growth density increased,more easily clustered group,increased and extended extension;In the experimental group(group C、D、E、F),the cell growth was relatively slow,not easy into the group,cell body shrink.(2)The proliferation of rat HSC-T6 was detected by CCK-8 method:The OD value of rat HSC-T6 was significantly higher in acetaldehyde group(P <0.01,Vs blank control group A,the experimental group C、D、E、F).The OD value of the experimental group(C、D、E、F)decreased gradually with the increase of LY294002 concentration,and the difference was statistically significant(P <0.01).(3)The expression of PI3 K and p-Akt was detected by Western-blotting:Compared with the blank control group,the relative expression of PI3 K and p-Akt protein in acetaldehyde group and acetaldehyde+LY294002 group were increased(P <0.01).Compared with acetaldehyde group,the expression of PI3 K and p-Akt protein in acetaldehyde + LY294002 group were decreased(P <0.01,P<0.05).Conclusions:(1)Acetaldehyde can stimulate the proliferation of rat HSC-T6,and the specific inhibitor of PI3 K /Akt pathway LY294002 can inhibit the proliferation of rat HSC-T6.(2)Liver injury and liver fibrosis caused by ethanol and its metabolites acetaldehyde may be have a relationship with PI3 K / Akt pathway.