Apocynin reduced doxycycline-induced acute liver injury in ovariectomized mice

Satoru Mitazaki; Midori Hashimoto; Yui Matsuhashi; Shigeyoshi Honma; Miwako Suto; Naho Kato; Kouichi Hiraiwa; Makoto Yoshida; Sumiko Abe

doi:10.1016/j.toxrep.2016.02.003

Apocynin reduced doxycycline-induced acute liver injury in ovariectomized mice

Toxicol Rep. 2016 Mar 5:3:357-363. doi: 10.1016/j.toxrep.2016.02.003. eCollection 2016.

Authors

Satoru Mitazaki¹, Midori Hashimoto¹, Yui Matsuhashi¹, Shigeyoshi Honma², Miwako Suto³, Naho Kato³, Kouichi Hiraiwa³, Makoto Yoshida², Sumiko Abe¹

Affiliations

¹ Laboratory of Forensic Toxicology, Faculty of Pharmacy, Takasaki University of Health and Welfare, 60 Nakaorui-machi, Takasaki 370-0033, Japan.
² Laboratory of Pathophysiology, Faculty of Pharmacy, Takasaki University of Health and Welfare, 60 Nakaorui-machi, Takasaki 370-0033, Japan.
³ Department of Legal Medicine, Fukushima Medical University, 1 Hikarigaoka, Fukushima 960-1295, Japan.

Abstract

To determine the physiological role of estrogen in the development of liver injury, we examined the sensitivities of sham and ovariectomy (ovx) mice against doxycycline (DOXY)-induced acute liver injury. Ovx or sham operation was performed in C57BL/6J wild-type female mice of eight weeks of age. Sham mice and ovx mice were treated with DOXY (240 mg/kg ip) 8 weeks after the operation, 30 min after apocynin (5 mg/kg) or saline administration. Blood and liver samples were obtained at 3 and 6 h after DOXY administration. Liver dysfunction occurred soon after DOXY administration and became more severe in ovx mice than in sham mice. At early phase after DOXY injection, TNF-α and iNOS inductions upregulated almost the same levels in sham and ovx mice. On the other hand, expression levels of IL-6, IL-10, c-fos, cox-2 and HO-1, downstream genes of TNF-α, were significantly increased in ovx mice compared to those in sham mice, correlated with liver dysfunction. In addition, apocynin, a NADPH oxidase (Nox) inhibitor, totally improved DOXY-induced liver injury in both sham and ovx mice, indicating that reactive oxygen species generated through Nox activation by DOXY are responsible for development of acute liver injury.

Keywords: ALF, acute liver failure; ALT, alanine aminotransferase; ARF, acute renal failure; Apocynin; DOXY, doxycycline; Doxycycline-induced liver injury; HO-1, heme oxygenase-1; IL-6, interleukin-6; NADPH oxidase; Nox, NADPH oxidase; Ovariectmized; Ovx, ovariectomy; ROS, reactive oxygen species; SOD, superoxide dismutase; STAT3, signal transducers and activators of transcription-3; TNF-α, tumor necrosis factor-α; cox-2, cyclooxygenase-2; iNOS, inducible nitric oxide synthase.