支持细胞TM4中金属硫蛋白1M和1G对锌响应的差异基因表达。
Differential gene-expression of metallothionein 1M and 1G in response to zinc in sertoli TM4 cells.
作者信息
Kheradmand Fatemeh, Nourmohammadi Issa, Modarressi Mohammad Hossein, Firoozrai Mohsen, Ahmadi-Faghih Mohammad Amin
机构信息
Dept. of Biochemistry, Iran University of Medical Sciences, Tehran, Iran.
出版信息
Iran Biomed J. 2010 Jan-Apr;14(1-2):9-15.
BACKGROUND
Zinc (Zn) as an important trace element is essential for testicular development and spermatogenesis. Molecular mechanism of Zn action in the reproductive system may be related to metal binding low-molecular weight proteins, metallothioneins (MT). Our objective was to determine the effect of Zn on two important isoforms of MT, MT1M and MT1G genes expression on testicular sertoli cells.
METHODS
Cultured sertoli TM4 cells were exposed to different concentrations of Zn at different time points. Cellular uptake of Zn was tested using flame atomic absorption spectrometry. The cellular viability and gene expression were assessed by MTT and real-time PCR methods, respectively.
RESULTS
The treated cells resulted in higher Zn concentration and cellular viability. The expression of MT1M and MT1G genes in the treated cells were greater than those of the untreated cells (P less than 0.05). In the high dosage treated group (100 and 500 muM), Zn concentration and expression of MT1M and MT1G genes increased three h after treatment; MT1G gene expression increased more at sixth h. At 18th h of treatment, the expression of both genes especially MT1G, increased dramatically while Zn concentration decreased.
CONCLUSION
Since the increase of MT1G mRNA was coincident with cellular Zn level, it seems that MT1G has a more prominent role than MT1M in the homeostasis of Zn. In addition, Zn at dosage of 50 muM (pharmacologic concentration) may protect cells by increasing the expression of MT genes at longer periods.
背景
锌（Zn）作为一种重要的微量元素，对睾丸发育和精子发生至关重要。锌在生殖系统中的作用分子机制可能与金属结合低分子量蛋白质——金属硫蛋白（MT）有关。我们的目的是确定锌对睾丸支持细胞中MT的两种重要亚型MT1M和MT1G基因表达的影响。
方法
将培养的支持细胞TM4在不同时间点暴露于不同浓度的锌。使用火焰原子吸收光谱法检测细胞对锌的摄取。分别通过MTT和实时PCR方法评估细胞活力和基因表达。
结果
处理后的细胞导致更高的锌浓度和细胞活力。处理后细胞中MT1M和MT1G基因的表达高于未处理细胞（P小于0.05）。在高剂量处理组（100和500μM）中，处理后3小时锌浓度以及MT1M和MT1G基因的表达增加；MT1G基因表达在第6小时增加更多。在处理后第18小时，两个基因尤其是MT1G的表达急剧增加，而锌浓度下降。
结论
由于MT1G mRNA的增加与细胞锌水平一致，似乎MT1G在锌的稳态中比MT1M发挥更突出的作用。此外，50μM（药理浓度）的锌可能通过在较长时间内增加MT基因的表达来保护细胞。
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