HGPAEs vector-based RNAi inhibits the expression of gene encoding MyD88 in rat liver tissues

Abstract

Objective To investigate the inhibitory effects of histidine grafted poly (β-amino ester) (HGPAEs) vector-based RNA interference (RNAi) on the expression of gene encoding myeloid differentiation factor 88 (MyD88) in rat liver tissues. Methods The sequence of small hairpin RNA (shRNA) was designed based on the genetic information of MyD88. HGPAEs vector was constructed and coupled with shRNA plasmid targeting MyD88 to construct pMyD88-HGPAEs vector. Rats were divided into five groups including control group, HGPAEs treatment group, pHK-HGPAEs treatment group, shRNA treatment group and pMyD88-HGPAEs treatment group. The rats in each group were transfected with the corresponding interventions through portal vein injection. Real-time PCR and Western blot assay were performed to detect the expression of MyD88 in liver tissues 3 days after transfection. Results The pMyD88-HGPAEs vector was successfully constructed. The expression of gene encoding MyD88 was inhibited in rats from shRNA treatment group and pMyD88-HGPAEs treatment group (P<0.05). Significantly decreased expression of gene encoding MyD88 at mRNA and protein levels were observed in rats from pMyD88-HGPAEs treatment group as compared with those from other groups (P<0.01). Conclusion HGPAEs vector might be used as a potential gene carrier. The expression of gene encoding MyD88 in rat liver tissues could be significantly inhibited through portal vein injection of pMyD88-HGPAEs vector. This study provided evidences for further research on pMyD88-HGPAEs vector in a high responder model of rat orthotopic liver transplantation. Key words: Histidine grafted poly (β-amino ester); Myeloid differentiation factor 88; RNA interference