Abstract: Objective:To investigate the effects of cold preservation on the expression of GATA in intrahepatic bile duct.
Methods The intrahepatic bile duct tissues of SD rats were obtained by collagenase perfusion combined with mechanical separation. After being cut into fragments, the intrahepatic bile duct tissues were cultured in rat tail collagen gel for 48 hours before experiment. All the rats were divided into the control group, cold preservation 1 hour (CP1 h) group and cold preservation 12 hours (CP12 h) group. There were 5 rats in each group. The mRNA and protein expressions of GATA were detected by RealTime polymerase chain reaction and Western blot. Measurement data with normal distribution were presented as ±s. Comparison among 3 groups was done by ANOVA and pairwise comparisons were done by LSD test.
Results:The mRNA expressions of GATA3, GATA4, GATA6 were detected, while the mRNA expressions of GATA1, GATA2 and GATA5 were undetectable in intrahepatic bile duct tissue of the control group. The mRNA expressions of GATA4 in the CP1 h group, CP12 h group and the control group were 0.72±0.08, 0.56±0.07 and 0.96±0.06, with significant difference among the 3 groups (F=38.981, P<0.05). The mRNA expression of GATA4 in the CP12 h group was significantly lower than that in the CP1 h group and the control group, and the mRNA expression of GATA4 in the CP1 h group was significantly lower than that in the control group (P<0.05). The mRNA expression of GATA6 in the CP1 h group, CP12 h group and the control group were 0.83±0.07, 0.68±0.12 and 0.98± 0.12, with significant difference among the 3 groups (F=10.175, P<0.05). The mRNA expression of GATA6 in the CP12 group was significantly lower than that in the CP1 h group and the control group, and the mRNA expression of GATA6 in the CP1 h group was significantly lower than that in the control group (P<0.05). The mRNA expressions of GATA3 in the CP1 h group, CP12 h group and the control group were 0.92±0.06, 0.89±0.05 and 0.98± 0.11, with no significant difference among the 3 groups (F=1.674, P>0.05). The protein expressions of GATA4 in the CP1 h group, CP12 h group and the control group were 0.78±0.07, 0.64±0.06 and 0.99±0.10, with significant difference among the 3 groups (F=24.211, P<0.05). The protein expression of GATA4 in the CP12 h group was significantly lower than that in the CP1 h group and the control group, and the protein expression of GATA4 in the CP1 h group was significantly lower than that in the control group (P<0.05). The protein expressions of GATA6 in the CP1 h group, CP12 h group and the control group were 0.90±0.04, 0.75±0.06 and 0.98±0.11, with significant difference among the 3 groups (F=11.651, P<0.05).  The protein expression of GATA6 in the CP12 h group was significantly lower than that in the CP1 h group and the control group (P<0.05).
Conclusion:The expressions of GATA4 and GATA6 in the intrahepatic bile duct tissues are decreased significantly after cold preservation, which indicate that GATA4 and GATA6 might be involved in the pathophysiological process of the bile duct after cold preservation.