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AIM: To investigate the role of nuclear factor- κB (NF- κB) in the expression of monocyte chemoatractant protein- 1 (MCP- 1) in human mesangial cells (HMCs) induced by oxidized low- density lipoprotein (Ox- LDL).METHODS: HMCs were used as target cells. Inhibitory κBα (IκBα) and MCP- 1 protein level was measured by cell ELISA.Activities of transcriptional factors NF- κB were determined by electrophoresis mobility shift assay (EMSA). Immunohistochemistry was used to detect the translocation of Rel p65. RESULTS: NF - κB DNA - binding activation in MCs was observed when 10-100 mg/L Ox - LDL was added to the medium, and 50 mg/L Ox - LDL caused the strongest effect (8.50 ± 1.14, P ＜ 0.01vs control; P ＜ 0.05 vs 10, 25 and 100 mg/L Ox - LDL). The most optimal stimulation time was 60 min ( 11.0 ± 2.11, P ＜0.01 vs control; P ＜ 0.05 vs 30 min or 240 min). IκBα protein level in MC dropped down most obviously after 60 min incubation with 50 mg/L Ox - LDL (0.050 ± 0.006, n = 5, P ＜ 0.01 vs control), while MCP- 1 expression level was the highest (0.331± 0.016, n = 5, P ＜ 0.01 vs control). The translocation of Rel p65 from cytoplasm to nucleus was detected too. NF - κB inhibitor pyrroledithiocarbomate (PDTC) could inhibit these effects induced by Ox- DL. CONCLUSION: Activation of NF- κB regulate the expression of MCP- 1 in HMCs induced by Ox - LDL.
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Objective To investigate the expressions and significance of protein kinase C (PKC) subtype α, β2 and ε in the pathogenesis of glomerulonephritis. Methods Immunohistochemical assays was used to examine the expressions of PKC α, PKC β2 and PKC ε in biopsy specimens of kidney from patients with mesangial proliferative glomerulonephritis (MsPGN) （n=16），nonproliferative golmerulonephritis (nonPGN) (n=12)，and in structurally normal kidneys (n=6). Results The PKC β2 expression in the glomerular was significantly higher in MsPGN patients than in nonPGN or normal kidney groups. Conclusion PKC β2 has a close association with MsPGN and might take part in the initiation of this renal disease.
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AIM: To investigate the role of nuclear factor-?B (NF-?B) in the expression of monocyte chemoatractant protein-1 (MCP-1) in human mesangial cells (HMCs) induced by oxidized low-density lipoprotein (Ox-LDL). METHODS: HMCs were used as target cells. Inhibitory ?B? (I?B?) and MCP-1 protein level was measured by cell ELISA. Activities of transcriptional factors NF-?B were determined by electrophoresis mobility shift assay (EMSA). Immunohistochemistry was used to detect the translocation of Rel p65. RESULTS: NF-?B DNA-binding activation in MCs was observed when 10-100 mg/L Ox-LDL was added to the medium, and 50 mg/L Ox-LDL caused the strongest effect (8.50?1.14, P
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Objective To investigate the role of nuclear factor-KB( NF-KB) in the expression of monocyte chemoattractant protein-i (MCP-1) in human mesangial cells induced by oxidized low density lipoprotein(Ox-LDL) . Methods Human mesangial cells were used as target cells. Levels of inhibitory KBK (IKB?) and MCP-1 protein were measured by ELISA. Activity of transcriptional factors NF-KB was determined by electrophoresis mobility shift assay ( EMSA) . Immunohistochemistry was used to detect the translocation of p65. Results Ox-LDL increased the DNA-binding activition of NF-KB and decreased the expression of IKB?. These effects were most obvious when the mesangial cells were cultured with Ox-LDL in 50 ?g/ml for 1 hour( P
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To explore the kinetics of LBP/CD14 system in patients with hemodialysis, and further to analyse its role in (he development of systemic inflammation response based on the changes of plasma endotaxin, TNF and IL-6 levels in patients with uremia treated with hemodilysis. Methods Sixteen patients with end-stage renal failure (8 cases with hemodialysis, 8 cses without hemodialysis) were selected for this study. limulus amebocyte lysate chromogenic assay, ELISA and cell in situ hybridization were used to determine the changes in endotoxin, LBP, TNF and IL-6 levels in plasma, and expression of CD14 mRNA in the monocytes. Results (l)Plasma LBP levels in patients with hemodialysis were significantly higher than those in patients without hemodialysis. The expression of CD14 mRNA in the monocytes in patients with hemodialysis was also more obvious. Both of them incresed much more in hemodialysis. (2)Plasma endotoxin levels in patients with hemodialysis, though being significantly higher than in patients without hemodialysis, were in low-level (61.7 ? 10.6 pg/ml). (3) Plasma TNF and IL-6 levels were markedly increased in patients with hemodialysis, which were significantly correlated with plasma LBP levels. Conclusion LBP/CD14 system in patients with hemodialysis is markedly up-regulated, which might be the important mechanism for low-level endotoxemia to exert its effects in hemodialysis.