ABSTRACT
Objective: To investigate the effects of tetrandrine (Tet) on expressions of proinflammatory and anti-inflammatory cytokine in lipopolysaccharide(LPS)- stimulated RAW264.7 cells. Methods: RAW 264.7 cells were cultured in media containing 1 μg/mL LPS and different doses of Tet. The proliferation of cells was examined by MTT assay, and nuclear translocation of NF-κB was detected and analyzed by the laser scanning confocal microscope (LSCM), and the levels of IL-6, TNF-α, and IL-10 were tested by ELISA. Results: Compared with the control group, Tet had no influence on the proliferation of RAW264.7 cells no matter with LPS or not when its concentration lower than 1 μmol/L; But when its concentration higher than 10 μmol/L, Tet significantly inhibited the growth of the cells whether LPS existed or not. Meanwhile, Tet significantly decreased the levels of IL-6, and TNF-α induced by LPS, and upregulated IL-10 in supernatant. At the mean time, results from LSCM showed that in Tet 1 μmol/L group, positive cells (dyeing-red p65 of NF-κB was translocated into nucleuses and made the nuclueses red) decreased siglificantly, while negative cells (dyeing-red p65 of NF-κB was collectted in endochylema and nucleuses became vacant) were predominantly founded. Conclusion: Tet has anti-inflammatory effect, which might be mediated by downregulating inflammatory factors IL-6, TNF-α through NF-κB signal pathway as well as increasing levels of anti-inflammatory factor IL-10.
ABSTRACT
The study is to investigate the effect of asiaticoside on collagen-induced arthritis (CIA). The model of CIA mice was prepared and the change of secondary paw swelling and the arthritis scores were observed. In vitro proliferation of spleen cells was examined using MTT assay. The cell-free protein extracts from the arthritic joints and nonarthritic joints were used for the analysis of protein expression of cyclooxygenase-2 (COX-2). And the level of PGE2 in joints was assayed using PGE2 express EIA kit. The tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels in the serum were measured by ELISA. Histopathological examination was performed by hematoxylin-eosin (HE) stain method. Asiaticoside (10, 20 and 40 mg x kg(-1) x d(-1), 22 d, ig) significantly reduced paw swelling, and decreased the arthritis scores. There was a significant reduction in proliferation of spleen cells of CIA mice treated with asiaticoside as compared with that of untreated CIA mice. COX-2, PGE2, TNF-alpha and IL-6 production in CIA mice were inhibited by asiaticoside. Meanwhile, the pathological examination showed that articular cartilage degeneration with synovial hyperplasia and inflammatory cells infiltration in CIA mice was suppressed by asiaticoside. Its active mechanism may be related to inhibiting proliferation of lymphocyte and reduction of expression of COX-2 and inflammatory cytokines.