Home > Browse Issues > Vol.29 No.2
Heme Oxygenase-1 Gene Silenced with siRNA to Reduce the Level of Bilirubin
Chun-E Li, You-Xing Jing1, Yi Shi1, LI-Qing Xu, Wen-Wei Zhong, Qing-Xiang Shen2, Yun-Zhu Li, Shan-Chang Yu, Zhen-Wei Xia1*
Department of Pediatrics, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; 1State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Scie
Abstract: Neonatal hyperbilirubinemia is a common clinical condition mainly caused by the increased production and decreased excretion of bilirubin. Current treatment is aimed at reducing serum level of bilirubin. Heme oxygenase (HO) is a rate-limiting enzyme which generates bilirubin. In this study， we intended to specifically suppress HO-1 using RNA interference technique. Small interfering RNA (siRNA)-1， -2 and -3 were designed based on human HO-1 (hHO-1) mRNA sequences. siRNA was transfected into a human hepatic cell line (HL-7702). hHO-1 transcription and protein levels were then determined. In addition， the inhibitory effect of siRNA on hHO-1 was assessed in the cells treated with hemin or transfected with hHO-1 plasmid. siRNA-3 showed the most potent suppressive effect on hHO-1. This inhibition is dose- and time- dependent. Compared with control， both hemin and hHO-1 plasmid up-regulated HO-1 expression in HL-7702 cells. However， the up-regulation was significantly attenuated by siRNA-3. Furthermore， the decrease of hHO-1 activity was coincident with the suppression of its transcription. Finally， siRNA-3 was shown to reduce hHO-1 enzymatic activity and bilirubin level. Thus， this study provides a novel therapeutic rationale via blocking bilirubin formation for preventing and treating neonatal hyperbilirubinemia as well as bilirubin encephalopathy at an early clinical stage.
CN
EN