Prolonged warm ischemia aggravates hepatic mitochondria damage and apoptosis in DCD liver by regulating Ca2+/CaM/CaMKII signaling pathway

Li Zhang; Sheng-Ning Zhang; Li Li; Xi-Bing Zhang; Rui-Chao Wu; Jun-Han Liu; Zhao-Yu Huang; Wang Li; Jiang-Hua Ran

Prolonged warm ischemia aggravates hepatic mitochondria damage and apoptosis in DCD liver by regulating Ca²⁺/CaM/CaMKII signaling pathway

Int J Clin Exp Pathol. 2019 Jan 1;12(1):217-228. eCollection 2019.

Authors

Li Zhang^{1

2}, Sheng-Ning Zhang¹, Li Li¹, Xi-Bing Zhang¹, Rui-Chao Wu¹, Jun-Han Liu¹, Zhao-Yu Huang¹, Wang Li¹, Jiang-Hua Ran¹

Affiliations

¹ Department of Hepatopancreatobiliary Surgery, The Affiliated Calmette Hospital of Kunming Medical University, The First People's Hospital of Kunming, Calmette Hospital Kunming, Yunnan Province, China.
² Department of Hepatopancreatobiliary, The First People's Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology Kunming, Yunnan Province, China.

PMID: 31933737
PMCID: PMC6944004

Abstract

This study was conducted to investigate the effect of warm ischemia duration on hepatocyte mitochondrial damage after liver transplantation, and confirm the role of CaMKIIγ in this process. Rat donation after cardiac death (DCD) liver transplantation model was established by exposing donor liver to 0 (W₀ group), 15 (W₁₅ group), and 30 (W₃₀ group) min warm ischemia. Some rats in W₁₅ group were transfected with CaMKIIγ and CaMKIIγ-shRNA lentivirus. On day 1, 3, and 7 post-transplantation, a series of experiments, including HE staining, TEM observation, ALT and AST measurement, flow cytometry analysis, qRT-PCR, and Western blotting were performed to evaluate the extent of hepatic and mitochondria damage. Within 7 days post-transplantation, prolonged ischemia led to an obvious deterioration of hepatic and mitochondria damage, presenting with a marked increase of apoptotic hepatocytes, ALT and AST levels, cells with low MMP, and AIF and Cyt C expression. CaMKIIγ overexpression caused the significant ultrastructural damage of hepatic cells, increase of cells with low MMP, enhancement of AIF and Cyt C expression, and augmented Ca²⁺/CaM/CaMKIIγ, while blocking CaMKIIγ showed an opposite result. In conclusion, ischemia duration is proportional to the extent of hepatic mitochondria damage, and CaMKIIγ plays a negative regulatory role in this process by regulating the Ca²⁺/CaM/CaMKII signaling pathway.

Keywords: Ca2+/CaM/CaMKII; DCD liver; Warm ischemia; mitochondria damage.