ABSTRACT
Objective: To study the effect of Bushen Zhuangjintang and glucosamine hydrochloride on the repairing of rat knee cartilage defect, the combined application of traditional Chinese and western medicine can promote the repairing of knee cartilage defect more effectively. To provide a new theoretical basis and method for the treatment of knee cartilage injury. Method: SPF rats (64 rats) were randomly divided into 5 groups, sham group,model group, Bushen Zhuangjintang group (7.5 g·kg-1·d-1), and glucosamine hydrochloride group(7.5 g·kg-1·d-1), Bushen Zhuangjin Decoction combined with glucosamine hydrochloride group[(7.5+7.5) g·kg-1·d-1] was administered to rats by drug gavage. The knee cartilage defects were repaired by gross observation and scanning electron microscopy at 4, 8 and 12 weeks. Real-time PCR was used to detect the expression of Collagen Ⅱ and Aggrecan mRNA in each group. Western blot was used to detect the expression of Collagen Ⅱ. Result: 4,8,12 weeks compared with group, Bushen Zhuangjintang combined with hyaluronic acid in the glucosamine hydrochloride group was filled with hyaline cartilage. The cartilage in the drilled area was smooth and smooth, and integrated with the surrounding cartilage tissue, which was superior to other groups. Real\|time PCR and Western blot analysis showed that the expression of Aggrecan protein and mRNA were significantly increased at 4, 8 and 12 weeks compared with the blank group (PPConclusion: The combined application of traditional Chinese and western medicine can obviously promote the repair of cartilage defects in the knee of rats. The possible mechanism of the treatment of cartilage injury by the combination of traditional Chinese and western medicine is analyzed from the level of gene protein expression and microstructure.
ABSTRACT
Objective To investigate the effect of recombinant adenovirus mediatied human endostatin (rAD-GFP-ES)on rats with collagen typeⅡinduced arthritis(CIA),and explore the mechanism of inflamma- tion and cytokines inhibition on rats CIA.Methods The rAD-GFP-ES was amplified and purified.The model of rat CIA was induced by intradermal injection of typeⅡcollagen combined with complete Freund's adjuvant(CFA). On the second day after the injection,the therapeutic administration of rAD-GFP-ES(1?10~(11)pfu?kg~(-1)?week~(-1)?4 weeks)were performed to the rats.The mean arthritis index(AI)was scored every week since then.The relative concentrations of ES,IL-I?,TNF-?in sera collected at the fourth week were evaluated by western blotting. Results①The titer of the purified rAD-GFP-ES and rAD-GFP was 6.6?10~(12)pfu/ml and 4.8?10~(12)pfu/ml,re- spectively(A_(260nm)/A_(280nm)＞1.3).②The concentration of ES in sera of the group treated with rAD-GFP-ES was 2.4-lold higher compared to the normal group.③The mean arthritis index of the group treated with rAD-GFP- ES was much lower than that of the model group.The administration of rAD-GFP-ES could significantly de- creas the production of IL-1?and TNF-?in sera.Conclusions①The rAD-GFP-ES is efficiently expressed in vivo.②The rAD-GFP-ES has an inhibitory effect on the arthritis index of rat CIA.③IL-1?and TNF-?are involved in the pathogenesis of RA.The rAD-GFP-ES has an inhibitory effect on the expression of IL-1?and TNF-?in rat CIA.
ABSTRACT
Objective:To evaluate the impact of altered collagen Ⅱ(CⅡ) peptide(S268-270) on T cell activation.Methods:Altered CⅡ263-272 with G268,P269 and K270 consecutively substitution with A or G were synthesized using solid phase various concentrations were added to HLADR1 transfected APC.Expression of FITC stainning was analyzed by fluorescence-activated cell sorting,and the binding of altered CⅡ peptide to HLA-DR1 molecule on cell membrane was evaluated.Irridiated HLA-DR1 expressing APC was incubated with CⅡ263-272 or hemagglutinin(HA)306-318 and altered CⅡ peptides at various concentrations for 2 hours before T cells(3.19) were added.Supernatant was harvested and then added to IL-2 dependent CTLL cell.The cell proliferation was examined by methotetrazolium(MTT) method.Results:The altered CⅡ peptide and wild type of CⅡ263-272 were able to competitively bind to HLA-DR1 molecule expressed on cell membrane of transfected APC.CⅡ or HA induced T cell activation was significantly inhibited by the altered peptide S268-270.Conclusion:Altered CⅡ263-272 with G268,P269 and K270 consecutively substitutions with A or G inhibited CⅡ263-272 and HA306-318 induced T cell activation through competitively binding to HLA-DR1,suggesting a new approach in inhibition of T cell activation in RA.
ABSTRACT
To study and optimize the fermentation parameters for expressing human-like collagenⅡduring E. coli high-density fermentation. The effects of pH, temperature, dissolved oxygen and induction instant on the cell growth and human-like collagenⅡproduction were investigated to optimize the fermentation conditions. The results demonstrated that the following conditions were beneficial for cell growth and foreign gene expression, controlling pH in phase induction at 6.8 and initial pH at 6.5, maintaining fermentation temperature and dissolved oxygen concentration was controlled at 34?C and 20% respectively, and implementing induction at the later logarithmic growth phase. Under the optimized condition, the cell density and human-like collagenⅡyield could reach 88.4 g/L and 14.2 g/L, respectively.