Protective effect of eNOS overexpression against ischemia/reperfusion injury in small-for-size liver transplantation

Zhang,Bo; Liu,Qiu‑Hua; Zhou,Cui‑Jie; Hu,Ming‑Zheng; Qian,Hai‑Xin

doi:10.3892/etm.2016.3762

Protective effect of eNOS overexpression against ischemia/reperfusion injury in small-for-size liver transplantation

Authors:
- Bo Zhang
- Qiu‑Hua Liu
- Cui‑Jie Zhou
- Ming‑Zheng Hu
- Hai‑Xin Qian
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Affiliations: Department of General Surgery, Zhangjiagang Hospital Affiliated to Soochow University, Zhangjiagang, Jiangsu 215600, P.R. China, Institute of Hepatopancreatobiliary Surgery, Yichang Central People's Hospital, China Three Gorges University, Yichang, Hubei 443003, P.R. China
Published online on: September 30, 2016 https://doi.org/10.3892/etm.2016.3762
Pages: 3181-3188
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Ischemia/reperfusion (I/R) injury can occur during small-for-size liver transplantation, resulting in delayed graft function and decreased long‑term graft survival. The aim of the present study was to evaluate the effects of genetic overexpression of endothelial nitric oxide synthase (eNOS) in protecting hepatocytes against I/R injury in a rat model of small-for-size liver transplantation. L02 liver cells were transfected with the eNOS gene using an adenovirus (Ad‑eNOS). eNOS expression was detected using quantitative polymerase chain reaction and western blot analysis. To evaluate the effect of eNOS overexpression, L02 cells were placed in a hypoxic environment for 12 h and immediately transferred to an oxygen‑enriched atmosphere. For in vivo testing, rats pretreated with Ad‑eNOS or control underwent small‑for‑size liver transplantation. At 6 h after reperfusion, the bile quantity, serum transaminase and nitric oxide (NO) levels, and histological outcomes were evaluated. Cell apoptosis was assessed by flow cytometry or TUNEL assay. In vitro, Ad‑eNOS prevented apoptosis in L02 cells with an increase in the level of NO in culture supernatant. In vivo, Ad‑eNOS pre‑treatment significantly increased bile production, improved abnormal transaminase levels, diminished apoptosis among liver cells, and decreased hepatocellular damage at 6 h after I/R injury. The eNOS‑mediated renal protective effects might be associated with the downregulation of tumor necrosis factor‑α and a reduction in macrophage activation in the early stage of reperfusion in small‑for‑size liver allografts. eNOS‑derived NO production significantly attenuates hepatic I/R injury. Thus, eNOS overexpression constitutes a promising therapeutic approach to prevent liver I/R injury following small‑for‑size liver transplantation.

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