Upregulated expression of hypoxia reactive genes in peripheral blood mononuclear cells from chronic liver disease patients

Akifumi Kuwano; Masatake Tanaka; Hideo Suzuki; Miho Kurokawa; Koji Imoto; Shigeki Tashiro; Takeshi Goya; Motoyuki Kohjima; Masaki Kato; Yoshihiro Ogawa

doi:10.1016/j.bbrep.2021.101068

Upregulated expression of hypoxia reactive genes in peripheral blood mononuclear cells from chronic liver disease patients

Biochem Biophys Rep. 2021 Jul 14:27:101068. doi: 10.1016/j.bbrep.2021.101068. eCollection 2021 Sep.

Authors

Affiliations

¹ Department of Medicine and Bioregulatory Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.
² Department of Hepatology, Iizuka Hospital, 3-83 Yoshio-machi, Iizuka, Fukuoka, 820-8505, Japan.
³ CREST, Japan Agency for Medical Research and Development, 1-7-1 Otemachi, Chiyoda-ku, Tokyo, 100-0004, Japan.

Abstract

Liver fibrosis induces intrahepatic microcirculation disorder and hypoxic stress. Hypoxic stress has the potential for an increase in the possibility of more liver fibrosis and carcinogenesis. Liver biopsy is a standard method that evaluates of intrahepatic hypoxia, however, is invasive and has a risk of bleeding as a complication. Here, we investigated the hypoxia reactive gene expressions in peripheral blood mononuclear cells (PBMC) from chronic liver disease patients to evaluate intrahepatic hypoxia in a non-invasive manner. The subjects enrolled for this study were composed of 20 healthy volunteers (HV) and 48 patients with chronic liver disease (CLD). CLD patients contained 24 patients with chronic hepatitis（CH）and 24 patients with liver cirrhosis (LC). PBMC were isolated from heparinized peripheral blood samples. We measured the transcriptional expression of hypoxia reactive genes and inflammatory cytokines by quantitative RT-PCR. mRNA expression of adrenomedullin (AM), vascular endothelial growth factor A (VEGFA) superoxide dismutase (SOD), glutathione peroxidase (GPx) (p < 0.05), Interleukin-6 (IL-6), transforming growth factor-beta (TGF-β) and heme oxygenase-1 (HO-1) in CLD group were significantly higher than HV. AM mRNA expression is correlated with serum lactate dehydrogenase (LDH), serum albumin (Alb), IL6, and SOD mRNA expression. The hypoxia reactive gene expression in PBMCs from CLD patients was more upregulated than HV. Especially, angiogenic genes were notably upregulated and correlated with liver fibrosis. Here, we suggest that mRNA expression of AM in PBMCs could be the biomarker of intrahepatic hypoxia.

Keywords: AM, Adrenomedullin; ANGPTL4, Angiopoietin-like 4; Adrenomedullin; CH, chronic hepatitis; CLD, chronic liver disease; Chronic liver disease; GPx, glutathione peroxidase; HCC, hepatocellular carcinoma; HCV, hepatitis C virus; HIF, hypoxia inducible factor; HO-1, heme oxygenase -1; HV, healthy volunteers; IL-6, Interleukin-6; Intrahepatic hypoxia; LC, liver cirrhosis; LDH, lactate dehydrogenase; MCP-1, Monocyte chemoattractant protein-1; PBMC, Peripheral blood mononuclear cells; PT, prothrombin time; Peripheral blood mononuclear cells; ROS, reactive oxygen species; SOD, Superoxide dismutase; TGF-β, transforming growth factor-beta; TNF-α, Tumor Necrosis Factor-α; VEGF, vascular endothelial growth factor; VEGFA, vascular endothelial growth factor A; VEGFR2, vascular endothelial growth factor receptor 2.