| Polysaccharides of Arnebia euchroma, Bupleurum chinense DC., Houttuynia cordata Thunb., Eucommia ulmoides Oliv. are the major bioactive molecules. Polysaccharides possess various biological activities, such as immune regulation, anti-infection and anti-tumoral activity.We investigated the effects of Arnebia euchroma Polysaccharides(AEP), Bupleurum chinense DC. Polysaccharides(BCP), Houttuynia cordata Thunb. Polysaccharides(HCP), Eucommia ulmoides Oliv. Polysaccharides(EOP) on TNF-α, IL-10 and IL-18 related cytokines mRNA expression in PBMC or LPS-stimulated PBMC. The transcription levels of TNF-α, IL-10, IFNy, IL-4, IL-18, IL-18BP, IL-18Rαand IL-18Rβwere detected by real-time RT-PCR. The phosphorylation of ERK in PBMC was detected by Western-blot.In this study, we have found AEP have anti-inflammatory and immunostimulatory function. Besides, we investigated AEP may inhibit LPS-induced TNF-αexpression in LPS-stimulated PBMC via down-regulation of MAPK signaling pathway.1. Effects of Polysaccharides on TNF-αand IL-18 related cytokines mRNA expression levels in PBMCTo investigate the effect of the AEP, BCP, HCP and EOP extracts on the production of proinflammatory cytokines, including TNF-α, IL-10 and IL-18 related cytokines, PBMC were cultured with AEP, BCP, HCP and EOP extracts. We found AEP, BCP, HCP and EOP induced the transcription of TNF-α, IL-10 and IL-18, but inhibited the transcription of IL-18BP in PBMC.We analyzed TNF-α, IL-10 and IL-18 related cytokines mRNA levels in PBMC stimulated with AEP, BCP, HCPand EOP extracts and compared the kinetics of cytokines expression at differen time points. As results shown, AEP induced transcription of TNF-αmRNA in PBMC, peaking at 2h, about 3.9 fold increasing in contrast with untreated cells. BCP, HCP and EOP also induced transcription of TNF-a mRNA. There was a significant increase by 2.5,5.3 and 3.3 fold in IL-10 production at 6h following incubation with BCP, AEP and HCP as compared with untreated cells. The transcription of IFNy was enhanced by 4.6,5.7 and 5.2 fold following AEP, BCP and HCP stimulation respectively, peaking at 6h. There was a gradual increase in the gene expressions for IL-4, starting as early as 1 h and peaking at 6 h following AEP and HCP stimulation, about 2.8 and 3.5 fold as compared with untreated cells. BCP induced transcription of IL-4, peaking at 2 h, with about 3.9 fold increasing in contrast with untreated cells. There was a gradual increase in the gene expressions for IL-18, peaking at 6 h following AEP stimulation, about 3.3 fold as compared with untreated cells. BCP induced transcription of IL-18, peaking at 2 h, with about 2.1 fold increasing in contrast with untreated cells. BCP and HCP inhibited transcription of IL-18BP in PBMC. The transcription of IL-18Rα, IL-18Rβin PBMC was induced by AEP, peaking at 6h.The results indicate that polysaccharides have anti-inflammatory and immunostimulatory function. It may serve as an effective therapeutic approach for inflammatory and autoimmune diseases.2. Effects of Arnebia euchroma Polysaccharides on inflammation-related cytokines mRNA expression levels in LPS-stimulated PBMCWe also investigated the effects of AEP on inflammation-related cytokines mRNA expression levels in LPS-stimulated PBMC. AEP inhibited transcription of TNF-αand IL-18 mRNA at 4h in LPS-stimulated PBMC (inhibition ratio,15-50%).To explore the possible signaling pathway. the effect of AEP on LPS-induced activation of MAPK signaling pathway was examined. The immunoblot assay showed that AEP inhibited LPS-induced phosphorylation of ERK (inhibition ratio,27%～ 61%). It suggests AEP may inhibit LPS-induced TNF-a expression in LPS-stimulated PBMC via down-regulation of MAPK signaling pathway.3. Staphylococcus carnosus as a novol vector for HBV DNA vaccineBesides, we investigated Staphylococcus carnosus as a novol vector for HBV DNA vaccine. First, we constructed pkmcp1/HBsAg plasmid, using a resistance gene and eukaryotic expression control sequence in eukaryotic expression vector pkmcp1. In this part, we observed the immune response induced by pkmcpl/HBsAg. To evaluate the HBsAg-specific antibody response, serum antibody titers were measured at different times after primary immunization. Mice immunized with pkmcp1/HBsAg developed low antibody at 4 week (1:200) after primary immunization, which were highly increased by the second challenge (1:900). At 8 weeks after the second challenge, anti-HBsAg titers decreaced at low levels (1:300). pkmcpl/HBsAg can express HBsAg after transformed into Staphylococcus Carnosus, and induce the production of anti-HBs antibodies. Our findings will contribute to the development of Staphylococcus Carnosus using in HBV DNA vaccine.
|
PDF Full Text Request