Gan Wei-hua, Din Gui-xia, Chen Rong-hua
Department of Pediatrics, the Second Affiliated Hospital, Nanjing 210011, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Sep;20(5):598-601.
To study the effect of oxidized low-density lipoprotein (Ox-LDL) on nuclear NF-kappaB activity in cultured glomerular mesangial cells (GMCs) from the Zucker rats and the correlation between the change of NF-kappaB activity and ras's age and genotype.
Four groups of GMCs (O(3m),O(10m), L(3m) and L(10m)) from the 3- and 10- month (fa/fa) and 3- and 10- month lean (Fa/Fa) Zucker rats were stimulated with Ox-LDL. Electrophoretic mobility shift assay (EMSA) and Western blot were used to detect the activity of NF-kappaB. Gel supershift assay(GSA) was used to detect the subunit of NF-kappaB dimer.
(1) NF-kappaB activity after induction with Ox-LDL in the 3 groups (O(3m),O(10m) and L(3m)) of GMCs were significantly higher than that in the control group (P<0.01). (2) With the increased of concentration and stimulation time of Ox-LDL, the NF-kappaB activity was increased correspondingly and reached the highest when GMCs were stimulated with 50 mg/L Ox-LDL for 4 hours. (3) Supershift assay demonstrated that Ox-LDL mainly activated the two subunits p65 and p50 in GMCs. (4) As the activity of NF-kappaB, O(3m) group vs O(10m) group, O(3m) group vs L(3m) group, and O(10m) group vs L(3m) group had all markedly differences (P<0.01), while the differences between L(3m) group vs L(10m) group was inferior to 3 groups above (P<0.05).
Ox-LDL could significantly activate NF-kappaB in GMCs from Zucker rats, in the time- and dose- dependence manner. There was a close relationship between the NF-kappaB activity in GMCs induced with Ox-LDL, the age and genotype of Zucker rats. NF-kappaB activity in GMCs of the Zucker rats induced with Ox-LDL plays an important role in the early renal lesion of diabetes mellitus.
研究氧化型低密度脂蛋白（Ox-LDL）对Zucker大鼠肾小球系膜细胞（GMCs）核内NF-κB活性的影响，以及NF-κB活性变化与大鼠年龄和基因型的关系。
用Ox-LDL刺激来自3个月和10个月龄（fa/fa）以及3个月和10个月龄瘦型（Fa/Fa）Zucker大鼠的四组GMCs（O(3m)、O(10m)、L(3m)和L(10m)）。采用电泳迁移率变动分析（EMSA）和蛋白质免疫印迹法检测NF-κB的活性。采用凝胶超迁移分析（GSA）检测NF-κB二聚体的亚基。
（1）三组GMCs（O(3m)、O(10m)和L(3m)）经Ox-LDL诱导后NF-κB活性显著高于对照组（P<0.01）。（2）随着Ox-LDL浓度和刺激时间增加，NF-κB活性相应升高，当GMCs用50mg/L Ox-LDL刺激4小时时达到最高。（3）超迁移分析表明，Ox-LDL主要激活GMCs中的两个亚基p65和p50。（4）作为NF-κB活性，O(3m)组与O(10m)组、O(3m)组与L(3m)组、O(10m)组与L(3m)组之间均有显著差异（P<0.01），而L(3m)组与L(10m)组之间的差异小于上述三组（P<0.05）。
Ox-LDL可显著激活Zucker大鼠GMCs中的NF-κB，呈时间和剂量依赖性。Ox-LDL诱导的GMCs中NF-κB活性与Zucker大鼠的年龄和基因型密切相关。Ox-LDL诱导的Zucker大鼠GMCs中NF-κB活性在糖尿病早期肾脏病变中起重要作用。
