Inhibiting the expression of uncoupling protein-2 attenuates acute damage to fatty liver cells / 中华器官移植杂志

ABSTRACT

Objective To investigate the effects of down-regulating uncoupling protein-2 (UCP-2) expression on acute damage to fatty liver cells and explore a new target for the donor liverwith steatosis. Methods Primary fatty liver cells were isolated from C57BL/6J-ob/ob transgenic miceby two-step collagenase perfusion method. RNAi lentivirus vector targeting mouse UCP-2 gene wasused to knock down the UCP-2 gene in the steatosis hepatocytes (the experimental group). Emptylentivirus vector was transfected into the steatosis hepatocytes cells as the control group. Under thefluorescence microscopy, the transfection efficiency was tested. Real time PCR was used to determinethe effect of RNAi. After the transfected cells were treated with TNF-α for 24 h, apoptosis wasanalyzed by flow cytometry using PI staining. Activation of caspase3 was detected by Western blot.Resalts The expression of UCP-2 gene was inhibited effectively, and the knockdown rate of UCP-2gene was 75%. The apoptosis rate in the experimental group was (4.97±0.25)%, significantlylower than in the control group [(21.13±1.28)%, p<0.05 ]. Activation 'of caspase3 in theexperimental group was also weaker than in the control group. Conclusion Inhibiting the expression ofUCP-2 can attenuate the injury of fatty liver cells.