Capacity of extracellular globins to reduce liver fibrosis via scavenging reactive oxygen species and promoting MMP-1 secretion
Vu Ngoc Hieu,
Le Thi Thanh Thuy,
Hoang Hai,
Ninh Quoc Dat,
Dinh Viet Hoang,
Ngo Vinh Hanh,
Dong Minh Phuong,
Truong Huu Hoang,
Hitomi Sawai,
Yoshitsugu Shiro,
Misako Sato-Matsubara,
Daisuke Oikawa,
Fuminori Tokunaga,
Katsutoshi Yoshizato,
Norifumi Kawada
Affiliations
Vu Ngoc Hieu
Department of Hepatology, Graduate School of Medicine, Osaka City University, Osaka, Japan
Le Thi Thanh Thuy
Department of Hepatology, Graduate School of Medicine, Osaka City University, Osaka, Japan
Hoang Hai
Department of Hepatology, Graduate School of Medicine, Osaka City University, Osaka, Japan
Ninh Quoc Dat
Department of Pediatrics, Hanoi Medical University, Hanoi, Viet Nam
Dinh Viet Hoang
Department of Hepatology, Graduate School of Medicine, Osaka City University, Osaka, Japan; Cho Ray Hospital, Ho Chi Minh City, Viet Nam
Ngo Vinh Hanh
Department of Hepatology, Graduate School of Medicine, Osaka City University, Osaka, Japan
Dong Minh Phuong
Department of Hepatology, Graduate School of Medicine, Osaka City University, Osaka, Japan
Truong Huu Hoang
Department of Hepatology, Graduate School of Medicine, Osaka City University, Osaka, Japan
Hitomi Sawai
Graduate School of Life Science, University of Hyogo, Hyogo, Japan
Yoshitsugu Shiro
Graduate School of Life Science, University of Hyogo, Hyogo, Japan
Misako Sato-Matsubara
Department of Hepatology, Graduate School of Medicine, Osaka City University, Osaka, Japan
Daisuke Oikawa
Department of Pathobiochemistry, Graduate School of Medicine, Osaka City University, Osaka, Japan
Fuminori Tokunaga
Department of Pathobiochemistry, Graduate School of Medicine, Osaka City University, Osaka, Japan
Katsutoshi Yoshizato
Academic Advisor's Office, PhoenixBio Co. Ltd., Hiroshima, Japan; Endowed Laboratory of Synthetic Biology, Graduate School of Medicine, Osaka City University, Osaka, Japan
Norifumi Kawada
Department of Hepatology, Graduate School of Medicine, Osaka City University, Osaka, Japan; Corresponding author. Department of Hepatology, Graduate School of Medicine, Osaka City University, 1-4-3 Asahimachi, Abeno, Osaka, 545-8585, Japan.
Background & aims: Hepatic stellate cells (HSCs) are the primary cell type in liver fibrosis, a significant global health care burden. Cytoglobin (CYGB), a globin family member expressed in HSCs, inhibits HSC activation and reduces collagen production. We studied the antifibrotic properties of globin family members hemoglobin (HB), myoglobin (MB), and neuroglobin (NGB) in comparison with CYGB. Approach & results: We characterized the biological activities of globins in cultured human HSCs (HHSteCs) and their effects on carbon tetrachloride (CCl4)-induced cirrhosis in mice. All globins demonstrated greater antioxidant capacity than glutathione in cell-free systems. Cellular fractionation revealed endocytosis of extracellular MB, NGB, and CYGB, but not HB; endocytosed globins localized to intracellular membranous, cytoplasmic, and cytoskeletal fractions. MB, NGB, and CYGB, but not HB, scavenged reactive oxygen species generated spontaneously or stimulated by H2O2 or transforming growth factor β1 in HHSteCs and reduced collagen 1A1 production via suppressing COL1A1 promoter activity. Disulfide bond-mutant NGB displayed decreased heme and superoxide scavenging activity and reduced collagen inhibitory capacity. RNA sequencing of MB- and NGB-treated HHSteCs revealed downregulation of extracellular matrix–encoding and fibrosis-related genes and HSC deactivation markers. Upregulation of matrix metalloproteinase (MMP)-1 was observed following MB and NGB treatment, and MMP-1 knockdown partially reversed globin-mediated effects on secreted collagen. Importantly, administration of MB, NGB, and CYGB suppressed CCl4-induced mouse liver fibrosis. Conclusions: These findings revealed unexpected roles for MB and NGB in deactivating HSCs and inhibiting liver fibrosis development, suggesting that globin therapy may represent a new strategy for combating fibrotic liver disease.
