PMID: 9164988May 1, 1997Paper

Competitive reverse transcriptase-polymerase chain reaction shows that dietary zinc supplementation in humans increases monocyte metallothionein mRNA levels

The Journal of Nutrition
V K Sullivan, Robert J Cousins

Abstract

Zinc status is difficult to evaluate in humans. Metallothionein gene expression is transcriptionally regulated by dietary zinc and thus could serve as an assessment parameter based on zinc-dependent function. We used semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) to establish that MT mRNA is increased in a human monocytic cell line by addition of zinc to the medium. To examine this response in human subjects, a dietary supplement of 50 mg zinc gluconate/d was given for 15 d. Monocytes were purified from venous blood using NycoPrep 1.068. Monocyte purity was determined by flow cytometry using fluorescent anti-human monocyte CD14 antibodies. Total monocyte RNA was extracted and converted to cDNA by reverse transcription. Competitive RT-PCR was used to analyze differences between cDNA levels that are proportional to MT mRNA levels in monocytes from zinc-supplemented and control subjects. RT-PCR oligonucleotide primers were designed to amplify both a 201 bp segment of the human MT cDNA and a 180 bp competitor cDNA template. The 180 bp competitor cDNA template was used for MT cDNA quantitation. The RT-PCR data show that there was a significant increase in monocyte MT mRNA in subjects within 6 d of zinc sup...Continue Reading

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Citations

Jul 10, 1998·The Proceedings of the Nutrition Society·R J Cousins
Apr 11, 2001·Journal of the American College of Nutrition·N R GlickW N Adkins
Aug 31, 1999·The Proceedings of the Nutrition Society·M L Failla
Aug 15, 2001·American Journal of Physiology. Cell Physiology·J C FanzoK Y Lei
Sep 14, 1999·The American Journal of Physiology·J Y WuK Y Lei
Jan 16, 2007·Journal of Animal Physiology and Animal Nutrition·D CarlsonH D Poulsen
May 8, 2009·The American Journal of Clinical Nutrition·Nicola M LoweTamás Decsi
Jul 11, 2002·American Journal of Physiology. Cell Physiology·Jessica C FanzoKai Y Lei
Nov 9, 2007·Food and Nutrition Bulletin·Sonja Y HessKenneth H Brown
Aug 15, 2000·The International Journal of Biochemistry & Cell Biology·N L BarnesE J Cornish
Jul 4, 2002·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·Xu-Fang LiangHiromi Oku
Jul 2, 2019·Journal of the American Chemical Society·Prem N BasaShawn C Burdette

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