Open hepatic artery flow with portal vein clamping protects against bile duct injury compared to pringles maneuver

Abstract

Background

Conventional hepatic artery and portal vein clamping strategies can prevent blood loss and ischemia-reperfusion liver injury, and such preventative measures are the key to successful liver surgery. However, ischemic-induced damage to cholangiocytes is rarely considered. Here, we aimed to investigate the effect of different hepatic inflow interruption methods on bile duct injury.

Methods

Forty rats were randomly grouped as sham, Pringle maneuver (PM) and hepatic arterial blood flow open (HAFO) groups. We evaluated liver histology and function in liver sections, and biliary histology, cholangiocyte apoptosis and proliferation, cytokine production, and bile composition. RNA sequencing is performed to explore possible molecular mechanisms. The Blood-biliary barrier permeability and tight junctions were analyzed by HRP injection, immunofluorescence staining and analysis of ZO-1 expression by immunoblotting.

Results

HAFO significantly attenuated ischemia-induced liver injury and decreased ALT, ALP, TBIL, and DBIL levels in serum. The histopathological observations showed that bile duct injury in the PM group was more serious than that in the HAFO group. The numbers of apoptotic biliary epithelial cells in HAFO-treated rat bile ducta were lower than those in the PM group. RNA-seq showed that tight junctions may be related to the mechanism underlying the protective effect of HAFO, as shown by the reduced HRP levels and increased ZO-1 and claudin-1/3 expression in the HAFO group compared to the PM group.

Conclusion

Compared with PM, HAFO alleviated the ischemic injury to the biliary system, which was characterized by decreased biliary epithelial cell apoptosis, reduced inflammatory responses and decreased blood-biliary-barrier permeability.

Keywords:

• Pringle maneuver
• hepatic artery
• biliary system
• ischemia injury

Author contributions

All authors contributed to this study and participated in the writing and drafting process as well as the critical review the manuscript. YB and LD conceived the work; BPD, WM designed the experiments; ZSL, CPL performed the experiments and analyzed the data; ZSL, YB and LD significantly contributed to the writing process. All authors have given the final approval of the version to be published and agreement to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

Ethical approval

The Animal Care and Use Committee of Kunming Medical University approved the study protocol (License: kmmu2021058). Euthanasia of animals was carried out according to the Chinese Code for the Care and Use of Animals for Scientific Purposes 8th edition.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by the National Natural Science Foundation of China [81860121]; Yunnan Applied Basic Research Projects-Kunming Medical University Union Foundation [202001AY070001-040]; Yunnan Provincial Fund for High Level Reserve Talents in Health Science [H-2018068].