Rdh13 deficiency weakens carbon tetrachloride-induced liver injury by regulating Spot14 and Cyp2e1 expression levels

doi:10.1007/s11684-017-0568-x

Front. Med.

2019, Vol. 13

Issue (1) : 104-111 https://doi.org/10.1007/s11684-017-0568-x

RESEARCH ARTICLE

Rdh13 deficiency weakens carbon tetrachloride-induced liver injury by regulating Spot14 and Cyp2e1 expression levels

Xiaofang Cui^1,², Benting Ma¹, Yan Wang³, Yan Chen¹, Chunling Shen¹, Ying Kuang², Jian Fei², Lungen Lu³(

), Zhugang Wang^1,²(

)

¹. State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
². Shanghai Research Center for Model Organisms, Shanghai 201203, China
³. Department of Gastroenterology, Shanghai First People’s Hospital, Shanghai Jiao Tong University, Shanghai 200080, China

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Abstract

Mitochondrion-localized retinol dehydrogenase 13 (Rdh13) is a short-chain dehydrogenase/reductase involved in vitamin A metabolism in both humans and mice. We previously generated Rdh13 knockout mice and showed that Rdh13 deficiency causes severe acute retinal light damage. In this study, considering that Rdh13 is highly expressed in mouse liver, we further evaluated the potential effect of Rdh13 on liver injury induced by carbon tetrachloride (CCl₄). Although Rdh13 deficiency showed no significant effect on liver histology and physiological functions under regular culture, the Rdh13^−/− mice displayed an attenuated response to CCl₄-induced liver injury. Their livers also exhibited less histological changes and contained lower levels of liver-related metabolism enzymes compared with the livers of wild-type (WT) mice. Furthermore, the Rdh13^−/− mice had Rdh13 deficiency and thus their liver cells were protected from apoptosis, and the quantity of their proliferative cells became lower than that in WT after CCl₄ exposure. The ablation of Rdh13 gene decreased the expression levels of thyroid hormone-inducible nuclear protein 14 (Spot14) and cytochrome P450 (Cyp2e1) in the liver, especially after CCl₄ treatment for 48 h. These data suggested that the alleviated liver damage induced by CCl₄ in Rdh13^−/− mice was caused by Cyp2e1 enzymes, which promoted reductive CCl₄ metabolism by altering the status of thyroxine metabolism. This result further implicated Rdh13 as a potential drug target in preventing chemically induced liver injury.

Keywords retinol dehydrogenase 13 carbon tetrachloride acute liver injury Cyp2e1 Spot14

Corresponding Author(s): Lungen Lu,Zhugang Wang

Just Accepted Date: 25 December 2017 Online First Date: 16 April 2018 Issue Date: 12 March 2019

Cite this article:

Xiaofang Cui,Benting Ma,Yan Wang, et al. Rdh13 deficiency weakens carbon tetrachloride-induced liver injury by regulating Spot14 and Cyp2e1 expression levels[J]. Front. Med., 2019, 13(1): 104-111.

URL:

https://academic.hep.com.cn/fmd/EN/10.1007/s11684-017-0568-x
https://academic.hep.com.cn/fmd/EN/Y2019/V13/I1/104

Fig.1 Liver functions and histology remain normal in Rdh13^−/− mice. (A) Serum levels of alanine aminotransferase, aspartate aminotransferase, and albumin are shown as mean values±S.E.M. from sex- and age-matched wild type and Rdh13^−/− mice (n = 6, P>0.05). (B) Histological analyses of liver sections stained with H&E are shown at original magnifications of 100× and 400×, correspondingly.

Fig.2 Acute liver injury induced by CCl₄ is alleviated in Rdh13^−/− mice. (A) Serum ALT levels in WT and Rdh13^−/− mice at the indicated times post-CCl₄ exposure are expressed as mean±S.E.M. (n = 4, *P<0.01, **P<0.05). (B) Representative H&E-stained liver sections of WT and Rdh13^−/− mice at the indicated times post-CCl₄ exposure are shown at an original magnification of 100×.

Fig.3 Apoptosis induced by CCl₄ treatment is reduced in Rdh13^−/− livers. (A) In situ fluorescence TUNEL assay on liver frozen sections shows hepatocyte nuclei (blue, 4′,6-diamidino-2-phenylindole) and apoptotic cells (green, fluorescein isothiocyanate). (B) Quantitative analysis on apoptotic cells in the liver upon CCl₄ treatment for the indicated time points (*P<0.05, **P<0.01). (C) Caspase-3 expression by immunohistochemistry in the liver upon CCl₄ treatment for the indicated time points. (D) Western blot analyses for the indicated proteins in the livers of mice exposed to CCl₄ for the indicated time points. Glyceraldehyde 3-phosphate dehydrogenase is shown as loading control.

Fig.4 Rdh13 deficiency reduced hepatocyte compensatory proliferation after CCl₄-induced liver damage in mice. (A) In vivo Brdu incorporation assayed by immunohistochemistry in the liver upon CCl₄ treatment for the indicated time points. (B) Percentages of Brdu-positive cells in the livers are shown as mean±S.E.M. (***P<0.001). (C) Cyclin D1 mRNA expression at the indicated time points in the livers by real-time RT-PCR is shown as mean±S.E.M. (*P<0.05, ** P<0.01). Protein levels of cyclin D1 in the livers after CCl₄ treatment are shown by immunohistochemistry (D) and Western blot (E). (F) Western blot for p-ERK1/2 and total ERK1/2 in the livers of WT and Rdh13^−/− mice after CCl₄ treatment is shown.

Fig.5 Decreased expression levels of Cyp2e1 and Spot14 in the livers of Rdh13^−/− mice after CCl₄ treatment. (A) Expression levels of Cyp2e1 mRNA and protein by real-time PCR (mean±S.E.M., ** P<0.01) and Western blot (B) show reduced Cyp2e1 expression levels in Rdh13^−/− livers 24 h after CCl₄ exposure. The expression levels of Spot14 in the livers after CCl₄ treatment are detected by quantitative real-time PCR (C, mean±S.E.M., * P<0.05, ** P<0.01) and Western blot (D).

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