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Aim To investigate the effect of phillygenin ( PHI) on lipopolysacchride ( LPS) and normal human plasma ( NHP) induced inflammatory injury on alveolar type II epithelial A549 cells and the related mechanism. Methods A549 cells were exposured to 1 mg • L
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Objective To study the antiendotoxin activity of P1 and P2 based on the lipopolysacchride binding protein.Method P1 and P2 were designed and obtained.In vitro test,peripheral blood mononuclear cell(PBMC)were extracted from volun-teer 100ml venous blood,the experiment group was arranged as:control group,positive control group,LPS group,LPS+P1 (2 mg/L,5 mg/L,12.5 mg/L)group and LPS+P2(2 mg/L,5 mg/L,12.5 mg/L)group,TNF-α and IL-6 of supernatant liquor in every group were detectd by ELISA.In vivo,40 kunming rice were randomly divided into four groups with ten rice each group:control group,model group,LPS+P1 and LPS+P2 group,Pathologic changes of lung and liver tissues were ob-served by hematoxylin and eosin(HE)staining.Results The serum level of IL-6 and TNF-αin 12.5 mg/L P1 and 2 mg/L, 5 mg/L,12.5 mg/L P2 treatment group were lower than that in model group,the difference was statistically significant(all P<0.01).Serum level of TNF-αor IL-6 in 12.5 mg/L P2 treatment group were similar to that in PMB treatment group, and there was no statistically significant difference(P>0.05).Histologymorphology finding showed that central veins of liver and hepatic sinusoid congestion,hepatic cellular edema existed,occasionally,acidophilic change and spotty necrosis were found,pulmonary interstitial edema,focal hemorrhage,alveolar space stenosis existed.As regards 10 mg/kg P1 treatment group mice,hepatic cellular edema and pulmonary interstitial edema ameliorated.About 10 mg/kg P2 treatment group mice, veins of liver and hepatic sinusoid congestion obviously ameliorated,mild pulmonary interstitial edema exsited.Conclusion The results indicated P1 and P2 had antiendotoxin effect,in vivo and vitro,for 12.5 mg/L P2,its inhibition effect for TNF-αor IL-6 release was positive.
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Aim To study the mechanisms of the pro-tective effect of procyanidin B2 ( PCB2 ) on the myocar-dial cell apoptosis induced by lipopolysaccharide ( LPS) . Methods Using the primary culture rat myo-cardial cells, myocardial cell injury model was induced by LPS. PCB2 low, medium and high dose groups, were cultured with 6. 25 , 12. 5 , 25. 0 μmol · L-1 PCB2 respectively in DMEM medium for 24 h continu-ously. Myocardial cell survival rate was determined by MTT colorimetric method. Cardiacmyocyte NOX activi-ty was determined by lucigen chemiluminescence meth-od . Western blot analysis was used to detect myocardi-al NADPH oxidase p47phox expression. TUNEL method was used to detect apoptosis and flow cytometry was used to determine the content of myocardial cells ROS. Results Compared with control group, the cell dam-age induced by LPS group myocardial cell survival rate significantly decreased ( P <0. 01 ) , and myocardial cell NOX activity, p47phox expression, apoptotic cell number and ROS content were significantly increased (P<0. 01). PCB2 low, medium and high dose groups cell survival rates were significantly elevated, myocar-dial cell NOX activity and p47phox expression, apoptotic cell number and the ROS content decreased significant-ly in a dose-dependent manner ( P <0. 01 ) . Conclu-sion PCB2 protects myocardial cell apoptosis induced by LPS via inhibiting the expression of NADPH oxidase activation, p47phox expression and reactive oxygen spe-cies generation.
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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.
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OBJECTIVE: Nuclear factor kappa B (NF-kappa B) is a transcriptional factor in the expression of cyclooxygenase 2 (COX-2), the key enzyme in production of prostaglandins. The purpose of this study was to investigate the activation of NF-kappa B in human gestational tissues obtained from term pregnant women by various inducers. METHODS: Myometrium, chorion, and amnion were collected during cesarean section from term pregnant women not in labor. Cells from gestational tissues were isolated and cultured with Interleukin-1beta (IL-1beta), Tumor Necrosis Factor-alpha (TNF-alpha) or Lipopolysaccharide (LPS). The activation of NF-kappa B in cells of each tissues was measured by luciferase assay. RESULTS: Luciferase activity analysis showed significantly higher activity of NF-kappa B in myometrial cells treated with LPS, in chorion cells treated with IL-1beta and TNF-alpha, and in amnion cells treated with IL-1beta and LPS than control. CONCLUSION: In cells of gestational tissue at term pregnancy, the activation of NF-kappa B is cell-specific and various according to inducers.