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Analysis of Cadmium, Epigallocatechin Gallate, and Vitamin C Co-exposure on PC12 Cellular Mechanisms

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Abstract

Exposure to cadmium (Cd) is a risk factor to health impairments, wherein its cytotoxicity is attributed to induction of oxidative stress. Usage of anti-oxidants, however, can help lessen the damaging effects of Cd. The effect of Cd interaction with low concentration of dietary anti-oxidants, l-ascorbic acid and (−)-epigallocatechin gallate (EGCG), to PC12 cellular mechanisms was examined. The expected toxicity of Cd was observed on PC12 cells but addition of l-ascorbic acid ameliorated this effect. On the other hand, addition of EGCG was able to increase the cytotoxicity of Cd and to decrease the protective effect of l-ascorbic acid against Cd. Increase in LDH activity and decrease in free sulfhydryl levels indicated cell membrane damage and oxidative stress, respectively, in Cd- and EGCG-Cd-treated cells. Downregulation of pro-apoptotic proteins (pro-caspase-9, p53, and ERK1) was observed in cells treated with Cd alone and EGCG-Cd, while upregulation of autophagy-linked proteins (p62 and pBeclin1) was found on l-ascorbic acid–Cd combination treatments. These findings indicate that Cd causes cells to undergo an autophagy-enhanced cell death; low-concentration EGCG and l-ascorbic acid promotes cell survival individually; however, interaction of EGCG with Cd showed enhancement of Cd toxicity and antagonism of l-ascorbic acid efficiency.

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Abbreviations

Akt:

Protein kinase B

Bax:

BCl2-associated X apoptosis regulator

EGCG:

(−)-Epigallocatechin-3-gallate

ERK1:

Extracellular signal–regulated kinase-1

GSH:

Glutathione

LDH:

Lactate dehydrogenase

mTOR:

Mammalian target of rapamycin

PBS:

Phosphate buffer saline

ROS:

Reactive oxygen species

SEM:

Standard error of mean

SH:

Thiol group

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Authors and Affiliations

  1. Graduate School of Environmental Science, Hokkaido University, Sapporo, 060-0810, Japan

    Serene Ezra C. Bondad & Masaaki Kurasaki

  2. Faculty of Environmental Earth Science, Hokkaido University, Sapporo, 060-0810, Japan

    Masaaki Kurasaki

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Bondad, S.E.C., Kurasaki, M. Analysis of Cadmium, Epigallocatechin Gallate, and Vitamin C Co-exposure on PC12 Cellular Mechanisms. Biol Trace Elem Res 198, 627–635 (2020). https://doi.org/10.1007/s12011-020-02097-9

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