摘要
AIM:To investigate the effect of NF-κB decoy oligodeoxynuleotides (ODNs) - modified dendritic cells (DCs) on regeneration of partial liver allograft.METHODS:Bone marrow (BM)- derived DCs from SD rats were propagated in the presence of GM-CSF or GM-CSF+IL-4 to obtain immature DCs or mature DCs, respectively. GMCSF-propagated DCs were treated with double-strand NF-κB decoy ODNs containing two NF-κB binding sites or scrambled ODNs. Allogeneic (SD rat to LEW rat) 50% partial liver transplantation was performed. Normal saline (group A),GM-CSF-propagated DCs (group B), GM-CSF+IL-4-propagated DCs (group C), and GM-CSF+NF-κB decoy ODNs(group D) or scrambled ODNs -propagated DCs (group E) were injected intravenously into recipient LEW rats 7 days prior to liver transplantation and immediately after transplantation.DNA synthesis (BrdU labeling) and apoptosis of hepatocytes were detected with immunostaining and TUNEL staining postoperative 24h, 48h, 72h and 84h,respectively. Liver graft-resident NK cell activity, hepatic IFN-γ mRNA expression and recipient serum IFN-γ level at the time of the maximal liver allograft regeneration were measured with ^51Cr release assay, semiquantitative RT-PCR and ELISA, respectively.RESULTS: Regeneration of liver allograft was markedly promoted by NF-κB decoy ODNs-modified immature DCs but was significantly suppressed by mature DCs, the DNA synthesis of hepatocytes peaked at postoperative 72h in group A, group B and group E rats, whereas the DNA synthesis of hepatocytes peaked at postoperative 84h in group C rats and 48h in group D rats, respectively. The maximal BrdU labeling index of hepatocytes in group D rats was significantly higher than that in the other groups rats.NF-κB decoy ODNs-modified immature DCs markedly suppressed but mature DCs markedly promoted apoptosis of hepatocytes, liver-resident NK cell activity, hepatic IFN-γ mRNA expression and recipient serum IFN-γ production.At the time of the maximal regeneration of liver allograft,the minimal apoptosis of hepatocytes, the minimal activity of liver-resident NK cells, the minimal hepatic IFN-γ mRNA expression and serum IFN-y production were detected in group D rats. The apoptotic index of hepatocytes, the activityof liver- resident NK cells, the hepatic TFN-γ mRNA expression level and the serum IFN-γ level in group D rats were significantly lower than that in the other groups rats at the time of the maximal regeneration of liver allograft.CONCLUSION:The data suggest that the augmented regeneration of partial liver allograft induced by NF-κB decoy ODNs-modified Des may be attributable to the reduced apoptotic hepatocytes, the suppressed activity of liverresident NK cells and the reduced IFN-γ production.
AIM:To investigate the effect of NF-κB decoy oligodeoxynuleotides(ODNs)-modified dendritic cells(DCs) on regeneration of partial liver allograft. METHODS:Bone marrow(BM)-derived DCs from SD rats were propagated in the presence of GM-CSF or GM-CSF+IL- 4 to obtain immature DCs or mature DCs,respectively.GM- CSF-propagated DCs were treated with double-strand NF-~B decoy ODNs containing two NF-κB binding sites or scrambled ODNs.Allogeneic(SD rat to LEW rat)50% partial liver transplantation was performed.Normal saline(group A), GM-CSF-propagated DCs(group B),GM-CSF+IL-4- propagated DCs(group C),and GM-CSF+NF-κB decoy ODNs (group D)or scrambled ODNs-propagated DCs(group E) were injected intravenously into recipient LEW rats 7 days prior to liver transplantation and immediately after transplantation.DNA synthesis(BrdU labeling)and apoptosis of hepatocytes were detected with immunostaining and TUNEL staining postoperative 24 h,48 h,72 h and 84 h, respectively.Liver graft-resident NK cell activity,hepatic IFN-γ mRNA expression and recipient serum IFN-γ level at the time of the maximal liver allograft regeneration were measured with ^(51)Cr release assay,semiquantitative RT-PCR and ELISA,respectively. RESULTS:Regeneration of liver allograft was markedly promoted by NF-κB decoy ODNs-modified immature DCs but was significantly suppressed by mature DCs,the DNA synthesis of hepatocytes peaked at postoperative 72 h in group A,group B and group E rats,whereas the DNA synthesis of hepatocytes peaked at postoperative 84 h in group C rats and 48 h in group D rats,respectively.The maximal BrdU labeling index of hepatocytes in group D rats was significantly higher than that in the other groups rats. NF-κB decoy ODNs-modified immature DCs markedly suppressed but mature DCs markedly promoted apoptosis of hepatocytes,liver-resident NK cell activity,hepatic IFN~ mRNA expression and recipient serum IFN-γ production.At the time of the maximal regeneration of liver allograft,the minimal apoptosis of hepatocytes,the minimal activity of liver-resident NK cells,the minimal hepatic IFN-γ mRNA expression and serum IFN-γ production were detected in group D rats.The apoptotic index of hepatooltes,the activity of liver-resident NK cells,the hepatic IFN-γ mRNA expression level and the serum IFN-γ level in group D rats were significantly lower than that in the other groups rats at the time of the maximal regeneration of liver allograft. CONCLUSION:The data suggest that the augmented regeneration of partial liver allograft induced by NF-κB decoy ODNs-modified DCs may be attributable to the reduced apoptotic hepatocytes,the suppressed activity of liver- resident NK cells and the reduced IFN-γ production.
基金
Supported by the Postdoctoral Science Foundation of China,No.2003033531
