The aim of this study was to investigate the subcellular localization and function of high density lipoprotein binding protein(HDLBP),and its relationship with the formation of goose fatty liver using in vivo and cellular models.Fourteen 70-day-old healthy Landes male geese were selected and raised in single cages.They were randomly divided into the control group(average weight 3.72 kg,ad libitum feeding)and the treating group(average weight 3.71 kg,overfed for 20 d)for in vivo experiment.Hepatocytes were isolated from 23-day-old Landes goose embryos and overexpressed with HDLBP gene for cellular experiment.Firstly,the subcellular localization of HDLBP protein in goose primary hepatocytes was determined by immunoblotting and immunofluorescence analyses.Secondly,the protein abundance of total HDLBP(wHDLBP)and mitochondrial HDLBP(mHDLBP)in the liver of the overfed geese and the control geese was determined by immunoblotting analysis.Then,HDLBP was overexpressed in goose primary hepatocytes,which was followed by determining the effects of HDLBP overexpression on the protein abundance of mHDLBP,the content of malondialdehyde(MDA),the activities of total superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-PX),the level of reactive oxygen species(ROS),and mitochondrial membrane potential.Finally,the differentially expressed genes(DEGs)and related signaling pathways affected by HDLBP overexpression were identified by transcriptome sequencing analysis,and some DEGs were verified in the in vivo model by quantitative polymerase chain reaction(PCR).The results showed that HDLBP could bind to mitochondria;the protein abundance of wHDLBP and mHDLBP in the overfeeding group was significantly lower than that in the control group(P＜0.01);overexpression of HDLBP in goose primary hepatocytes significantly increased the protein abundance of mHDLBP(P＜0.05),increased the levels of MDA(P＜0.01)and ROS(P＜0.05),and decreased the mitochondrial membrane potential(P＜0.05)and the activities of T-SOD(P＜0.05)and GSH-PX(P＜0.05).The up-regulated DEGs affected by HDLBP overexpression were mainly enriched in immune/inflammation-related pathways.In addition,compared with the control group,the expression of inflammation-related genes including IL1R1,TNFSF10,LTC4S,NCF1,SFTPA1,and KDR in the overfeeding group might be significantly reduced via HDLBP regulation(P＜0.05,0.01 or 0.001).HDLBP can bind to mitochondria,and overfeeding significantly reduced the protein levels of wHDLBP and mHDLBP in goose liver.Overexpression of HDLBP leads to mitochondrial dysfunction,oxidative stress and increased expression of inflammatory factors.Therefore,HDLBP may provide protection for goose fatty liver by affecting mitochondrial function,regulating oxidative stress and inflammatory response.
维普
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