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衰老标记蛋白-30/葡萄糖酸内酯酶在单个小鼠肝细胞中的年龄相关性降低:免疫组织化学和免疫荧光法

Age-associated decrease of senescence marker protein-30/gluconolactonase in individual mouse liver cells: Immunohistochemistry and immunofluorescence.

作者信息

Ishigami Akihito, Masutomi Hirofumi, Handa Setsuko, Maruyama Naoki

机构信息

Molecular Regulation of Aging, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan.

出版信息

Geriatr Gerontol Int. 2015 Jun;15(6):804-10. doi: 10.1111/ggi.12347. Epub 2014 Oct 14.

DOI:10.1111/ggi.12347
PMID:25311772
Abstract

AIM

Senescence marker protein-30 (SMP30)/gluconolactonase (GNL) is an age-associated protein in that its presence decreases with aging. Here, we used immunohistochemical analysis to investigate the changes of SMP30/GNL in individual cells of the liver from progressively aged mice.

METHODS

Male C57BL/6 strain mice at 1, 3, 6, 12, 24 and 30 months-of-age were the source of hepatic cells used to detect SMP30/GNL. Liver sections from these mice were subjected to immunohistochemical staining with anti-SMP30/GNL antibody. For immunofluorescent staining, primary cultured hepatocytes from mice at various ages were stained with SMP30/GNL and albumin.

RESULTS

In liver cells from mice of all ages, SMP30/GNL staining appeared in some but not all parenchymal cells, and localized in both the nuclei and cytoplasm. Moreover, SMP30/GNL-positive staining of parenchymal cells was present only around central vein areas, but not at sites of portal veins. Furthermore, the number of SMP30/GNL-positive cells increased as mice aged from 1 to 12 months, then decreased from the 12th to 24th month. Results were similar in primary cultured hepatocytes from mice of various ages.

CONCLUSIONS

SMP30/GNL-positive cells localized mainly around the central veins in the livers of mice and decreased numerically with aging, although there was no age-related change in counts of albumin-positive cells. SMP30/GNL protein occupied the nuclei and cytoplasm. Therefore, nuclear SMP30/GNL protein might be a regulatory factor specific for genes whose expression governs transcription and the aging process.

摘要

目的

衰老标志物蛋白30(SMP30)/葡萄糖酸内酯酶(GNL)是一种与年龄相关的蛋白,其含量随衰老而降低。在此,我们采用免疫组化分析方法,研究不同年龄阶段小鼠肝脏单个细胞中SMP30/GNL的变化情况。

方法

选取1、3、6、12、24和30月龄的雄性C57BL/6品系小鼠作为肝细胞来源,用于检测SMP30/GNL。对这些小鼠的肝脏切片进行抗SMP30/GNL抗体免疫组化染色。对于免疫荧光染色,用SMP30/GNL和白蛋白对不同年龄小鼠的原代培养肝细胞进行染色。

结果

在各年龄段小鼠的肝细胞中,SMP30/GNL染色出现在部分而非全部实质细胞中,且定位于细胞核和细胞质。此外,实质细胞的SMP30/GNL阳性染色仅出现在中央静脉区域周围,而门静脉部位未出现。而且,SMP30/GNL阳性细胞数量在小鼠1至12月龄时增加,随后在第12至24月龄时减少。不同年龄小鼠原代培养肝细胞的结果相似。

结论

SMP30/GNL阳性细胞主要定位于小鼠肝脏的中央静脉周围,且数量随衰老而减少,尽管白蛋白阳性细胞计数无年龄相关变化。SMP30/GNL蛋白存在于细胞核和细胞质中。因此,细胞核中的SMP30/GNL蛋白可能是一种特异性调节因子,作用于那些其表达调控转录和衰老过程的基因。

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