摘要
Background and Aims:Collagenβ(1-O)galactosyltransferase 25 domain 1(GLT25D1)is associated with collagen production and glycosylation,and its knockout in mice results in embryonic death.However,its role in liver fibrosis remains elusive,particularly in hepatic stellate cells(HSCs),the primary collagen-producing cells associated with liver fibrogenesis.Herein,we aimed to elucidate the role of GLT25D1 in HSCs.Methods:Bile duct ligation(BDL)-induced mouse liver fibrosis models,primary mouse HSCs(mHSCs),and transforming growth factor beta 1(TGF-β1)-stimulated LX-2 human hepatic stellate cells were used in in vivo and in vitro studies.Stable LX-2 cell lines with either GLT25D1 overexpression or knockdown were established using lentiviral transfection.RNA-seq was performed to investigate the genomic differences.HPLCMS/MS were used to identify glycosylation sites.Scanning electronic microscopy(SEM)and second-harmonic generation/two-photon excited fluorescence(SHG/TPEF)were used to image collagen fibril morphology.Results:GLT25D1 expression was upregulated in nonparenchymal cells in human cirrhotic liver tissues.Meanwhile,its knockdown attenuated collagen deposition in BDL-induced mouse liver fibrosis and inhibited mHSC activation.GLT25D1 was overexpressed in activated versus quiescence LX-2 cells and regulated in vitro LX-2 cell activation,including proliferation,contraction,and migration.GLT25D1 also significantly increased liver fibrogenic gene and protein expression.GLT25D1 upregulation promoted HSC activation and enhanced collagen expression through the TGF-β1/SMAD signaling pathway.Mass spectrometry showed that GLT25D1 regulated the glycosylation of collagen in HSCs,affecting the diameter of collagen fibers.Conclusions:Collectively,the upregulation of GLT25D1 in HSCs promoted the progression of liver fibrosis by affecting HSCs activation and collagen stability.
基金
funded by the National Science Foundation of China [No.82170541]
National Science Foundation of China [No.81900549]
Natural Science Foundation of Beijing Municipality [No.7202071]
The Capital Foundation for Clinical Characteristic Applied Research Projects [No.Z181100001718084]
The Digestive Medical Coordinated Development Center of Beijing Municipal Administration of Hospitals [No.XXZ0404]
The Study on Modernization of Traditional Chinese Medicine [No.2018YFC1705700]
Capital Medical University Research Development Fund [No.PYZ20031].
