| 1. The anti-inflammatory effect of plant polysacchridesAim:To determine anti-inflammatory effect of plant polysacchrides.Methods:(1)The determination of anti-complementary activity:The anti-complementary activity of crude polysaccharides from Arnebia euchroma(CAEP), Houttuynia cor data Thunb(CHCP)andBupleurum smithiiWolff var.parvifolium(BPs) by the classical pathway-mediated complement activation. (2)The effects of plant polysacchrides on cell viability and production of inflammatory mediators of macrophages:Murine peritoneal macrophages were randomly divided into 4 groups:control group, LPS (1μg/ml) stimulated group, polysacchrides treated groups, LPS combined with polysacchrides treated groups. Macrophages were treated for 24 h. Cell viability was measured by the MTT assay. And the levels of NO, TNF-a, and IL-lβ were respectively quantified by Griess reaction and enzyme-linked immunosorbent assay.(3)The effects of CHCP andBPs on virus induced lung injury:KM mice were randomly divided into 7 groups, including 4 kinds:normal group, virus model group, BPstreated group (20,40, 80mg/kg), CHCP treated group (20,40,80mg/kg). Chemotactic factors and inflammatory mediators in lung homogenate were quantified by ELISA to discover the effects of CHCP andBPson virus induced mice.Results:(1) The anti-inflammatory effect of CAEP.CAEP had a significant anti comlementaty activity (CH50=379±11 μg/ml). For peritoneal macrophages, CAEP alone promoted cell viability (P< 0.05), and increased the secretion of NO, TNF-a, and IL-1β at 100 μg/ml (P< 0.05). After LPS stimulation, the production of NO, TNF-a, and IL-1β released by peritoneal macrophages increased significantly (P< 0.001). CAEP obviously inhibited LPS-stimulated production of NO, TNF-a, and IL-1β (P< 0.05).For peritoneal macrophages, two pure polysaccharides from Arnebia euchroma had different activities. APS-1 inhibited LPS-stimulated production of NO(P< 0.05), and did not change the cell viability of peritoneal macrophages. APS-2 promoted the cell viability of peritoneal macrophages (P< 0.05), but it had no significant effect on LPS-stimulated secretion of NO.(2) The anti-inflammatory effect of BPsBPs had a significant anti-comlementaty activity (CH50=335±3 μg/ml). For peritoneal macrophages, BPs alone increased the secretion of NO, TNF-a, and IL-1β at 100 μg/ml (P< 0.05), and did not change the cell viability. After LPS stimulation, BPs obviously decreased LPS-stimulated production of NO, TNF-a, and IL-1β (P< 0.05).The pure polysaccharides from Bupleurum smithiiWolff var.parvifolium(BPPs) alone increased the production of NO released by peritoneal macrophages more sinificantly than BPs (P< 0.001). After LPS stimulation, BPPs obviously decreased the LPS-stimulated production of NO.In vivo, the levels of IL-6, IFN-a, MIP-1, and RANTES were increased significantly and the levels of IL-10 were decreased remarkably in the virus model group compared with the normal group (P< 0.01). BPs obviously inhibited the production of IL-6 and Rantes (P< 0.01), elevated the production of IL-10 (P< 0.01), but had no effects on IFN-a and MIP-1 levels.(3) The anti-inflammatory effect of CHCP.CHCP showed a high anti-comlementaty activity. The inhibition lysis of CHCP (300 μg/ml) were 75%. For peritoneal macrophages, CHCP significantly increased the secretion of NO, TNF-a, and IL-1β at 100 μg/ml (P< 0.05), and did not change the cell viability. After LPS stimulation, CHCPobviously decreased LPS-stimulated production of NO, TNF-a, and IL-1β(P<0.05).In vivo, the levels of IL-6, IFN-α, MIP-1, and RANTES were increased significantly and the levels of IL-10 were decreased remarkably in the virus model group compared with the normal group (P< 0.01). CHCP obviously inhibited the production of IL-6, MIP-1 and Rantes(P< 0.01), elevated the production of IL-10 (P <0.01), but had no significant effects on the level of IFN-α.(4) The anti-inflammatory effects of the other plant polysaccharidesAs for peritoneal macrophage:①The polysaccharides from bupleurum chinense DC(BCPs), prunella vulgaris(PVs), dryopteris setosa(DSs) significantlyenhanced theproduction of NO; BCPs, DSs, eucommia ulmoidespolysaccharides(Eus) enhanced the production of IL-1β;BCPs, Eus, PVs, DSs and Radixsophoraetonkinesis (RPs)promoted secretion of TNF-a except Sophora flavescens (SFs).②After LPS stimulation, BCPs, Eus, PVs,DDs,and RPssuppressed LPS-stimulated production of NO, TNF-a, and IL-1β. SFs had no effects on NO, and IL-1βproduction, but furthermore enhanced the level of TNF-a.Conclusion:(1)In vitro, we found that both CAEP, BPs and CHCP had significant anti-complementary activitiies and could inhibited the production of inflammatory mediators.. (2)BPs and CHCP relieved lung injury of virus-induced mice and inhibited production of some chemotactic factors and inflammatory mediators significantly.(3)Most polysacchridesalone promoted production of inflammatory mediators (NO, TNF-a, IL-1β) and inhibited them after LPS stimulation. It indicated there could be a similar effect on peritoneal macrophage among these polysacchrides.2. The benifical effects of TLE on systemic lupusAIM:This study was to determine whether the inhibitory oligodeoxynucleotide (INH-ODN) drug (TLE) had beneficial effects on experimental systemic lupus erythematosus.Methoths:BALB/c mice were immunized with CJ-S13 in Freund’s complete adjuvant on day 0, and then boosted on day 14 and day 43. TLE 5,10,20 or 40 μg/mousewere given to BALB/c mice subcutaneously from day 1 to day 55, three times a week. The index of spleen, thymus, popliteal lymph node and liver were measured. Serum samples of mice were tested by enzyme-linked immunosorbent assays (ELISA) for the presence of anti-dsDNA, anti-nuleosome antibodies and total IgG. Urine protein and creatinine were measured and kidneys were examined by light microscopy and graded by pathologist.Results:(1) Effect ofTLE on body weight and organ indexBoost injection on day 14 caused significant weight loss in CJ-S131 immunized mice compared with the untreated mice (P<0.05). TLE had no significant on weight loss (P>0.05). Boost injection on day 43 had no effects on the body weight of all mice. The index of popliteal lymph node almost at the same level. The index of spleen thymus and liver increased significantly in model group when compared with niormal group (P<0.05). TLE inhibited thymus swelling slightly (P> 0.05), but had no effects on the index of spleen and liver.(2) Effect of TLE on autoantibodies and total IgG productionThe anti-ds-DNA and anti-nucleosome antibodies levels were significantly elevated in the model group compared with the normal and model control group (P<0.001), and were reduced by TLE 20 or 40 μg/mouse significantly(P< 0.05).In the model control and model group, serum levels of total IgG were significantly increased compared with the normal group(P< 0.001),but TLE had no significant effect on production of total IgG.(3) Effect of TLE on renal histologyThe ratio of proteinuria and urine creatinine in the modelgroup was significantly higher than the normal and model control group (P<0.01). Treatment with TLE (10, 20,40μg/mouse) caused a marked reduction onratio compared with the model group (P<0.05).In the mice of model group, mesangial cell proliferation, expansion of the mesangial matrix and glomerular hypertrophy was observed. These lesions were not apparent in normal and model control group.Treatment withTLE (10,20,40μg/mouse) markedly ameliorated the glomerular injury.Conclusion:These findings suggested that inhibitory oligodeoxynucleotide had a beneficial effect on systemic lupus erythematosus-like syndroma induced by CJ-S131 in BALB/c mice. |
PDF Full Text Request