ABSTRACT
Objective To investigate the inhibitory effects of histidine grafted poly (β-amino es-ter) ( HGPAEs) vector-based RNA interference ( RNAi) on the expression of gene encoding myeloid differ-entiation factor 88 (MyD88) in rat liver tissues.Methods The sequence of small hairpin RNA (shRNA) was designed based on the genetic information of MyD 88.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 inter-ventions 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 MyD 88 was inhibited in rats from shRNA treat-ment 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 pMyD 88-HGPAEs treatment group as compared with those from other groups (P<0.01).Conclusion HGPAEs vector might be used as a po-tential gene carrier .The expression of gene encoding MyD 88 in rat liver tissues could be significantly inhibi-ted through portal vein injection of pMyD 88-HGPAEs vector .This study provided evidences for further re-search on pMyD88-HGPAEs vector in a high responder model of rat orthotopic liver transplantation .
ABSTRACT
AIM:To investigate the influence of high-mobility group box 1 (HMGB1) on the proliferation of neural stem cells in peri-infarction cortex of focal cerebral ischemia/reperfusion model rats .METHODS: Male SD rats (n=48) were randomly divided into sham group , ischemia/reperfusion (I/R) group, RNA interference group and nega-tive interference group .The rat middle cerebral artery was blocked to establish focal cerebral I /R model ( ischemia for 1 h and reperfusion for 7 d).Lentivirus vector of HMGB1 shRNA was used to suppress the HMGB1 protein expression in the rat brain.The effect of RNA interference was evaluated by the methods of double-immunofluorescence labeling of HMGB 1/GFAP and Western blotting .The proliferation of neural stem cells in the peri-infarction cortex was assessed by double labe-ling of BrdU/nestin.RESULTS: The protein expression of HMGB1 in I/R group was much higher than those in sham group (P<0.05).RNA interference effectively inhibited the HMGB1 expression (P<0.05).Double labeled BrdU/nestin positive cells in I/R group were more than that in sham group (P<0.05).The double labeled BrdU/nestin positive cells were significantly decreased in RNA interference group (P<0.05).CONCLUSION:Focal cerebral ischemia/reperfusion injury promotes the proliferation of neural stem cells in peri-infarction cortex by increasing HMGB 1 protein level .