Integrin ?V?5 Induces Hepatic Sinusoidal Capilarlization Via FAK/ERK Pathway Under High Glucose/oxLDL

Objective The main purpose of this study is to explore the relationship between integrin ?v?5,FAK,ERK,VN and dysfunction of HLSECs under high glucose/ ox-LDL conditions and to determine the mechanism on the indexes.Methods HLSECs were cultured and treated with media containing glucose(25m M,24h)/ox-LDL(100 ug/ml,24h)in the presence or absence of different inhibiors or not(?v?5 inhibitor RGDfv,ERK inhibitor DF228,FAK inhibitor PD98059).MTT colorimetry was used to detect the activity of HLSECs and expression of integrin ?v?5,FAK,ERK,VN were detected by RT-PCR,Western blot.Scanning electron microscopy was used to visualise the fenestration frequency and fenestration diameter.Expression of VN was also testified by immunofluorescence assay.All instructives were performed using the SPSS 21.0 software.Results 1.The expression of integrin ?v,integrin ?5,FAK,ERK,VN increased in a time-and oncentration-dependent manner under high glucose or ox LDL.Furthermore,It shpwed obvious inscreaing and strongest viability is at a glucose concentration of 25 m M,24 h and ox LDL concentration of 100 ?g/ml,24 h.This effect is the most significant when they co-exist(P<0.05).2.The expression of ERK,FAK and VN was down-regulated when HLSECs were treated with the integrin ?v?5 inhibitor RGDfv.DF228,inhibitor of FAK only suppressed expression of ERK and VN.Furthermore,VN expression was down-regulated by intervention of HLSECs with the ERK inhibitor PD98059,while the expression of integrin ?v?5 and FAK was not significantly changed(P<0.05).3.In order to examine whether high glucose and ox LDL interacts with the fenestrae of HLSECs.We used SEM at 5000× magnification to observe the porosity and number of fenestrae in ox LDL(100 ug/ml)and high glucose(25m M)treated HLSECs separately or jointly for 24 h.We observed that high glucose and ox LDL can lead to a decrease in the porosity,diameter and number of fenestrae compared with the control group,which is more significant in the combination of high glucose and ox LDL(P<0.05).Conclusion Integrin ?v?5 which induces HLSECs dysfunction,promoting hepatic sinusoidal capillarization regulates VN expression via FAK/ERK pathway under high glucose/ox-LDL,may be a potential target for prevention and treatment of diabetes with fatty liver disease.