Experimental study of compound BAM15 alleviating cryopreservation-induced primary hepatocyte injury in rats / 器官移植

ABSTRACT

Objective To investigate the effect of compound BAM15 on the primary hepatocyte injury induced by cold storage in rats. Methods The primary rat hepatocytes were extracted by collagenase perfusion method. According to different cell culture conditions, the cells were divided into 4 groups group A (Hibernate cell culture solution containing 250 nmol/L BAM15), group B (Hibernate cell culture solution containing 500 nmol/L BAM15), group C (Hibernate cell culture solution containing 1 000 nmol/ L BAM15), control group (Hibernate cell culture solution). The cells of each group were cryopreserved for 12 h. The purity of primary hepatocytes was observed under fluorescence microscope. The changes in the cell proliferation ability, cell apoptosis rate and mitochondrial reactive oxygen species (ROS) were measured in each group. Results The cell proliferation ability in groups B and C was significantly higher than that in the control group (both P < 0.05). The apoptosis rates in groups A, B and C were (33.7±2.2)%, (19.7±1.1)% and (28.7±1.2)%, which were significantly lower than (82.7±4.2)% in the control group (all P < 0.05). The positive rates of intracellular ROS in groups A, B and C were (11.8±4.0)%, (7.6±1.3)% and (8.9±1.6)%, remarkably lower than (27.4±4.5)% in the control group (all P < 0.05). Conclusions Compound BAM15 can effectively mitigate the primary hepatocyte injury in rats induced by cryopreservation. The underlying mechanism is probably associated with the role of BAM15 in reducing ROS generation during cold ischemia.