Changes associated with aging and replicative senescence in the regulation of transcription factor nuclear factor-kappa B
Changes associated with aging and replicative senescence in the regulation of transcription factor nuclear factor-kappa B
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DOI：
10.1042/bj3180603
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发表时间：
1996-09-01
影响因子：
4.1
通讯作者：
Salminen, A
中科院分区：
文献类型：
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作者：
Helenius, M;Hanninen, M;Salminen, A
动物的衰老和培养细胞的衰老都涉及基因表达模式的改变，表明转录因子调控的变化。我们通过电泳迁移率变动结合分析研究了转录因子核因子 (NF)-kappa B、激活蛋白因子-1 (AP-1) 和 Sp-1 与年龄相关的变化；我们还通过蛋白质印迹分析分析了 NF-κ B 复合物的蛋白质成分的变化。从年轻和老年 NMRI 小鼠和 Wistar 大鼠的心脏、肝脏、肾脏和大脑以及衰老前、衰老和猿猴病毒 40 永生化人 WI-38 成纤维细胞中制备细胞核和细胞质提取物。小鼠和大鼠的衰老导致所有研究组织中 NF-κ B 因子的核结合活性持续强烈增加，而 AP-1 和 Sp-1 的核结合活性则下降。在肝脏中。 NF-κ B 复合物的 p50、p52 和 p65 成分的蛋白质水平在细胞质部分中没有显示出任何与年龄相关的变化，但在细胞核部分中，p52 的水平在衰老过程中在心脏和肝脏中强烈增加。在所研究的任何组织中，抑制性 IκB-α 和 Bcl-3 成分的蛋白质水平均不受衰老影响。人 WI-38 成纤维细胞的复制性细胞衰老导致核 NF-κ B、AP-1 和 Sp-1 结合活性大幅下降。衰老WI-38成纤维细胞核部分中NF-κB复合物的p50和p52成分的蛋白水平降低，但衰老成纤维细胞细胞质中p65蛋白水平升高。细胞衰老还略微降低了 I kappa B-alpha 和 Bcl-3 的蛋白质水平。使用 NF-kappa B 增强子驱动的氯霉素乙酰转移酶报告基因转染测定显示，衰老 WI-38 成纤维细胞中 NF-kappa B 启动子活性显着下调。结果表明，组织和培养的成纤维细胞中的衰老过程可能受到不同的调节，这可能反映了有丝分裂细胞和有丝分裂后细胞之间的差异。在组织中，衰老似乎涉及 NF-κ B 成分调节的特定变化，也可能涉及 NF-κ B 复合物的 DNA 结合亲和力的变化。
Both the aging of animals and the senescence of cultured cells involve an altered pattern of gene expression, suggesting changes in transcription factor regulation. We studied age-related changes in transcription factors nuclear factor (NF)-kappa B, activator protein factor-1 (AP-1) and Sp-1 by using electrophoretic mobility shift binding assays; we also analysed changes in the protein components of NF-kappa B complex with Western blot assays. Nuclear and cytoplasmic extracts were prepared from heart, liver, kidney and brain of young adult and old NMRI mice and Wistar rats as well as from presenescent, senescent and simian virus 40-immortalized human WI-38 fibroblasts. Aging of both mice and rats induced a strong and consistent increase in the nuclear binding activity of NF-kappa B factor in all tissues studied, whereas those of AP-1 and Sp-1 decreased, e.g. in liver. Protein levels of p50, p52 and p65 components of the NF-kappa B complex did not show any age-associated changes in the cytoplasmic fraction but in the nuclear fraction the level of p52 strongly increased in heart and liver during aging. The protein levels of inhibitory I kappa B-alpha and Bcl-3 components were not affected by aging in any of the tissues studied. Replicative cellular senescence of human WI-38 fibroblasts induced a strong decrease in nuclear NF-kappa B, AP-1 and Sp-1 binding activities. Protein levels of p50 and p52 components of NF-kappa B complex were decreased in the nuclear fraction of senescent WI-38 fibroblasts but in the cytoplasm of senescent fibroblasts the level of p65 protein was increased. Cellular senescence also slightly decreased the protein levels of I kappa B-alpha and Bcl-3. Transfection assays with NF-kappa B-enhancer-driven chloramphenicol acetyltransferase reporter gene showed a significant down-regulation of NF-kappa B promoter activity in senescent WI-38 fibroblasts. Results suggest that the aging process might be regulated differently in tissues and cultured fibroblasts, perhaps reflecting differences between mitotic and post-mitotic cells. In tissues, aging seems to involve specific changes in the regulation of NF-kappa B components and perhaps also changes in the DNA-binding affinities of the NF-kappa B complex.