Effects of Autologous Bone Marrow Mononuclear Cells Implantation on Cholangiocytes Apoptosis in Model of Intrahepatic Ischemic Type Biliary Lesion in Rabbits

Abstract: Aim: To investigate the effects of autologous bone marrow mononuclear cells (BM-MNCs) implantation on regulation of cholangiocyte apoptosis in a model of intrahepatic ischemic type biliary lesion (ITBL) in rabbits. Materials and Methods: Thirty Japanese white rabbits were divided randomly into 3 groups (10 per group) including controls (group A), ITBL model (group B), and BM-MNCs implantation groups (group C). All rabbits underwent the same surgical procedure to prepare the liver for graft removal during transplantation. Subsequently, no additional vascular intervention was performed in group A. In group B, the hepatic artery and common bile duct were clamped with microvascular clips for 2 hours, where after the clips were removed to recover the blood supply. Group C received, BM-MNCs (108 cells per rabbit) injected through the hepatic artery after removing the clips. The animals were killed 4 weeks after operation. The survival rate, histopathologic examination, cholangiocyte apoptosis with terminal uridine nick-end labeling (TUNEL) staining and expressions of Bcl-2 and Bax proteins were examined using immunohistochemical staining. Results: Group A animals showed a survival of 100%; the rates in groups B and C were both 90%. Histopathologic examination revealed normal intrahepatic cholangiocytes in group A, obviously damaged ones in group B, and alleviated damage in group C. TUNEL staining indicated apoptosis of cholangiocytes in group B was more serious than that in group A or group C. Immunohistochemical staining demonstrated significantly decreased Bcl-2 expression in group B compared with that in group A; Bcl-2 expression in group C returned to the level of group A. Simultaneously, the Bax expression presented adverse results; the ratios of Bcl-2/Bax were ranked as group A > group C > group B. Conclusion: Implantation of autologous BM-MNCs significantly reduced apoptosis of intrahepatic cholangiocytes and prevented or abated intrahepatic ITBL. [Copyright &y& Elsevier]