LOX-1 Regulates Cardiac Fibroblasts Fibrosis Induced By High Glucose Through GSK-3β/STAT3 Pathway

Objective: To investigate whether plant hemagglutinin-like oxidized low-density lipoprotein receptor-1(LOX-1)regulates high glucose induced Cardiac fibroblasts(CFs)fibrosis through glycogen synthase kinase-3β(GSK-3β)/Signal transducer and activator of transcription 3(STAT3)pathway.Methods: CFs were isolated,cultured and identified.LOX-1 RNAi lentiviral vector was constructed to infect CFs.The experimental groups were as follows: Normal control(NC group),High glucose(HG group),LOX-1 silencing group(LV-LOX-1 group),Empty carrier group(LV-Con group).High glucose(25 m M,24 h),GSK-3β inhibitor SB216763 and STAT3 inhibitor STATTIC were used to intervene,and the activity of CFs was detected by CCK-8method.The mRAN and protein expression levels of LOX-1,Thioredoxin domain-containing protein5(TXNDC5),Collagen type I(COL-I),GSK-3β,STAT3,p-GSK-3β and p-STAT3 were detected by quantitative Real time-PCR and Western blotting.Results:1.CFs infected with LOX-1 RNAi lentiviral vector were obtained,which was green under fluorescence microscope.The mRNA and protein expression levels of LOX-1,COL-I,TXNDC5 in LV-LOX-1 group were decreased,Compared with the NC group,the mRNA expression levels of LOX-1,COL-I,TXNDC5,In LV-LOX-1 group were decreased by 49.8%,36.6% and 73.0%,and the protein expression levels were decreased by37.8%,57.9% and 85.0%,respectively(P < 0.01).2.Compared with NC group,The mRNA expression levels of LOX-1,COL-I and TXNDC5 in HG group were increased by 1.23 times,1.38 times,1.10 times,and the protein expression levels were increased by 44.0%,1.16 times,60.6%,respectively(P < 0.01).3.Compared with HG group,the mRNA and protein expression levels of LOX-1 in HG+LVLOX-1 group were decreased by 43.1% and 23.1%,the protein expression level of p-GSK-3β was increased by 71.1%,the protein expression level of p-STAT3 was decreased by 33.0%,the mRNA expression levels of COL-I and TXNDC5 were decreased by 33.7% and 30.8%,and the protein expression levels of COL-I and TXNDC5 were decreased by 21.5% and 36.0%,respectively(P <0.01).4.Compared with HG+LV-LOX-1group,the protein expression level of p-GSK-3β in HG+LV-LOX-1+SB216763 group was increased by 39.0%,and the protein expression level of pSTAT3 was decreased by 27.9%,the mRNA expression levels of COL-I and TXNDC5 were decreased by 24.9% and 20.1%,and the protein expression levels of COL-I and TXNDC5 were decreased by 17.3% and 27.4%,respectively(P < 0.01).5.Compared with HG+LV-LOX-1 group,the protein expression level of p-STAT3 in HG+LVLOX-1+STATTIC group was decreased by 51.4%,and the mRNA expression levels of COL-I and TXNDC5 were decreased by 48.8% and 47.2%,the protein expression level of COL-I and TXNDC5 were decreased by 35.2% and 38.2%,respectively(P < 0.01).Conclusion: LOX-1,GSK-3β,STAT3,COL-I and TXNDC5 are involved in high glucose induced CFs fibrosis.LOX-1 promotes the expression of COL-I and TXNDC through GSK-3β/STAT3 pathway,and inhibition of LOX-1 can inhibit high glucose induced CFs fibrosis.