Telomeric length and telomerase activity vary with age in peripheral blood cells obtained from normal individuals
Telomeric length and telomerase activity vary with age in peripheral blood cells obtained from normal individuals
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DOI：
10.1007/s004390050711
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发表时间：
1998-04-01
期刊：
影响因子：
5.3
通讯作者：
Shay, JW
中科院分区：
文献类型：
--
作者：
Iwama, H;Ohyashiki, K;Shay, JW
测量了124名4-95岁健康个体外周血单个核细胞的端粒酶活性和端粒长度。使用内部端粒酶测定标准、荧光引物和自动激光荧光 DNA 测序仪，通过荧光端粒重复扩增方案（荧光-TRAP）半定量评估端粒酶活性水平。通过末端限制性片段 (TRF) 测定测量端粒长度，并使用 (TTAGGG)(4) 探针通过 Southern 印迹分析在 HinfI 消化的 DNA 中确定端粒长度。对 80 名个体的 TRF 长度进行了测定，并观察到与年龄相关的尺寸逐渐减小。从正常个体（4-39岁）获得的外周血单核细胞中，TRF长度每年减少约84 bp，而在年龄大于或等于40岁的个体中，每年减少41 bp。相反，端粒酶活性随着衰老呈现出明显的双相模式。 4-39岁个体端粒酶活性逐渐下降，而40岁以上个体中有65%的端粒酶活性相对稳定但非常低，其余40岁以上个体未检测到端粒酶活性。
The telomerase activity and length of telomeres of peripheral blood mononuclear cells obtained from 124 healthy individuals aged 4-95 years was measured. Telomerase activity level was semiquantitatively assessed by a fluorescent-telomeric repeat amplification protocol (fluorescent-TRAP) using an internal telomerase assay standard, fluorescent primers and an automated laser fluorescent DNA sequencer. Telomeric length, measured by assay of terminal restriction fragments (TRFs), was determined in HinfI-digested DNA by Southern blot analysis using a (TTAGGG)(4) probe. TRF length was determined in 80 individuals and age-related progressive reduction of size was observed. TRF length in peripheral blood mononuclear cells obtained from normal individuals (aged 4-39 years) decreased by approximately 84 bp per year, while in individuals aged greater than or equal to 40 years it decreased by 41 bp per year. In contrast, telomerase activity showed an apparent biphasic pattern with aging. Individuals aged 4-39 years showed a progressive decrease in telomerase activity, whereas 65% of those aged greater than or equal to 40 years showed relatively stable but very low telomerase activity, and the remaining individuals aged greater than or equal to 40 years had no detectable telomerase activity.