摘要
目的 :探讨Akt/NF -κB信号途径在免疫复合物 (ICs)诱导肾小球系膜细胞 (MC)表达趋化因子中的作用。方法 :体外培养小鼠MC。对照组 :0 5 % (v/v)lipofectin作用 8h后IgG单体刺激 ;刺激组 :0 5 % (v/v)lipofectin作用 8h后AIgG(一种标准的ICs模型 )刺激 ;反义、正义、错义寡核苷酸组 :0 5 % (v/v)lipofectin转导Akt1反义、正义、错义寡核苷酸 8h后AIgG刺激。ELISA法检测单核细胞趋化蛋白 - 1(MCP - 1)、巨噬细胞集落刺激因子 (CSF - 1)浓度 ,mRNA半定量用RT -PCR。EMSA法检测NF -κB活性。结果 :MC有低水平NF -κB活化和组成性MCP - 1、CSF- 1mRNA及蛋白表达。AIgG刺激后 ,NF -κB活化增强 (0 35± 0 0 6vs 0 75± 0 16 ,P <0 0 1) ,MCP - 1和CSF - 1mRNA表达上调 (0 4 8± 0 0 3vs 0 72± 0 0 2 ,P <0 0 5 ;0 4 4± 0 0 1vs 0 5 9± 0 0 2 ,P <0 0 5 ) ,蛋白分泌增多 (15 5 2±1 81vs 4 3 0 5± 3 18,P <0 0 5 ;389 0 6± 13 75vs 76 4 2 2± 31 78,P <0 0 5 )。Akt1反义寡核苷酸显著抑制AIgG诱导NF -κB活化 (0 37± 0 0 5vs 0 75± 0 16 ,P <0 0 1)、MCP - 1和CSF - 1mRNA(0 5 2± 0 0 2vs 0 72± 0 0 2 ,P <0 0 5 ;0 4 4± 0 0 1vs 0 5 9± 0 0 2 ,P <0 0 5 )?
AIM: To explore the role of Akt/NF-κB pathway in immune-complexes-induced monocyte chemoattractant protein-1 (MCP-1) and colony stimulating factor-1 (CSF-1) expression in Mesangial Cells. METHODS: Primary murine glomerular mesangial cells were cultured in vitro and divided into control group, stimulation group and antisense, sense and mismatched oligodeoxynucleotide group. In control group, the cells were stimulated with monomeric IgG after treatment with 0.5% lipofectin for 8 h. In stimulation group, the cells, which had been treated with 0.5% lipofectin for 8 h, were stimulated with aggregated IgG. In antisense, sense and mismatched oligodeoxynucleotide group, being transduced antisense, sense and mismatched oligodeoxynucleotide respectively with 0.5% lipofectin 8 h, the cells were stimulated with AIgG. MCP-1 and CSF-1 in supernatant were deteced with ELISA. In addition, RT-PCR was used to determine MCP-1 and CSF-1 mRNA expression, and EMSA to investigated the activation of NF-κB. RESULTS: Mesangial cells cultured in vitro had a low level NF-κB activation and a low level constitutive expression of MCP-1 and CSF-1. Stimulated with AIgG, activation of NF-κB was markedly increased(0.35±0.06 vs 0.75±0.16, P<0.01), expression of MCP-1 and CSF-1 mRNA (0.48±0.03 vs 0.72±0.02, P<0.05; 0.44±0.01 vs 0.59±0.02, P<0.05), MCP-1 and CSF-1 levels in supernatant(15.52±1.81 vs 43.05±3.18, P<0.05; 389.06±13.75 vs 764.22±31.78, P<0.05) were markedly increased. Akt1 antisense oligodeoxynucleotide markedly inhibited immune-complexes-induced NF-κB activation, MCP-1 and CSF-1 mRNA and protein expression. CONCLUSION: Akt/NF-κB pathway mediates immune-complexes-induced MCP-1 and CSF-1 expression in mesangial cells. It suggests that Akt/NF-κB pathway may be a new therapy target for macrophage recruitment and activation in immune complexes nephritis.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第5期724-728,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目 (No .30 0 70 717)
教育部博士点基金资助项目 (No .2 0 0 0 0 5 6 916 )
