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目的 :研究氧化低密度脂蛋白 (Ox LDL)对体外培养的Zucker大鼠肾小球系膜细胞 (GMC)核因子κB (NF κB)活性的影响以及NF κB的活性变化与大鼠鼠龄及基因型的相关性。方法 :(1)将 3月龄和 10月龄Zucker肥胖大鼠及Zucker瘦型大鼠的GMC(O3m、O10m,、L3m和L10m)进行传代培养。 (2 )采用电泳迁移率变动分析 (EMSA)和超迁移率实验 (GSA) ,检测不同浓度及不同时间相的Ox LDL对GMC中NF κB活性的影响及其亚单位p5 0和p6 5的变化。利用Westernblot检测Ox LDL刺激后NF κB的核转位。结果 :Ox LDL刺激后 ,O3m、O10m及L3m3组GMC中NF κB的活性均明显强于对照组 (P <0 .0 1) ;L10m组与对照组相比较无显著差异 (P >0 .0 5 )。随着Ox LDL浓度的增加和刺激时间的延长 ,GMC中NF κB活性也相应增强 ,5 0mg/L的Ox LDL刺激 4h时 ,NF κB的活性强度达最高峰。Ox LDL主要激活NF κB的p6 5及p5 0亚单位 ;各组NF κB的活性相比较 :O3m 组高于O10m组 ,O3m组高于L3m组及O10m组高于L10m组 (P均 <0 .0 1) ;L3m组NF κB的活性高于L10m组 (P <0 .0 5 ) .结论 :Ox LDL可诱导Zucker大鼠GMC中NF κB活化 ,且呈时间和浓度依赖性 ;活性强度与大鼠的基因型及鼠龄密切相关。Ox LDL诱导的NF κB活化在Zucker肥胖大鼠的早期肾损害中起着更为重?
AIM: To investigate the effects of Ox LDL on the activity of nuclear factor κB (NF κB) in rat mesangial cells (GMC) cultured in vitro and the effect of oxidative low density lipoprotein Genotype correlations. Methods: (1) GMCs (O3m, O10m, L3m and L10m) were cultured in Zucker rats and Zucker mice at 3 months and 10 months of age. (2) EMSA and GSA were used to detect the effect of Ox LDL with different concentrations and time phases on NF κB activity in GMCs and the changes of its subunits p5 0 and p6 5 . Western blot was used to detect NF-κB nuclear translocation after Ox LDL stimulation. RESULTS: After Ox LDL stimulation, NFκB activity in GMCs in O3m, O10m and L3m3 groups was significantly higher than that in control group (P <0.01); there was no significant difference between L10m group and control group (P> 0.05) 5). With the increase of OxLDL concentration and stimulation time, the activity of NFκB in GMC increased correspondingly. The peak intensity of NFκB activity was reached when OxLDL was treated with 50mg / L of Ox LDL for 4h. Ox LDL mainly activated p65 and p50 subunits of NF-kappaB; The NF-κB activity in each group was higher than that in O10m group, O3m group and O10m group (P <0.05). (P <0.05) .Conclusion: Ox LDL can induce NF κB activation in GMCs in Zucker rats in a time and concentration-dependent manner. Rat genotypes and age are closely related. Ox LDL-induced activation of NF-κB plays a more important role in early kidney damage in Zucker obese rats?