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Protective Role of Metallothioneins in Human Endothethial Cells in Acute Response to High Dose Cadmium Chloride
Author(s) -
Zhang Yanmin,
Kucynda Theresa T.,
Marsboom Glenn,
KajdacsyBalla Andre
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.982.6
Subject(s) - cadmium chloride , reactive oxygen species , metallothionein , cadmium , umbilical vein , chemistry , cadmium exposure , flow cytometry , extracellular , andrology , oxygen , microbiology and biotechnology , pharmacology , medicine , biochemistry , biology , organic chemistry , in vitro
Cadmium is a well‐recognized environmental pollutant with numerous adverse health effects. Recent research has demonstrated the association of cadmium with cardiovascular disease through endothelial dysfunction at substantially low levels of chronic exposure. However, little is know about the effect of high concentration cadmium effect on human endothelial cells after acute exposure. Methods Primary human umbilical vein endothelial cells (HUVECs) were treated with varying doses of cadmium chloride (CdCl 2 ). Flow cytometry was used to evaluate the cells reactive oxygen species (ROS) with cell‐permeant 2′,7′‐dichlorodihydrofluorescein diacetate (H2DCFDA) probe. The oxygen consumption rate (OCR) of adherent cells was measured at 37°C using a Seahorse XF24 Extracellular Flux Analyzer. Gene expression was measured with real‐time RT‐PCR. Results The HUVECs cells were treated with varying dose of CdCl 2 for 24 hours and cells viability was not changed at any of the dose used. Real time RT‐PCR data revealed both metallothionein‐1A ( MT1A) and metallothionein‐2A ( MT2A) gene levels were significant increased by 50 μM CdCl 2 compared to the control levels (13.8‐fold and 5.4‐fold increase, respectively, P<0.05). OCR of HUVECs was decreased by 50 μM CdCl 2 treatments from 66.97±2.41 to 42.00±5.22 pMoles/min (P<0.005), and H2DCFDA fluorescence intensity was decreased from 2989±87.75 in control to 2165±54.75 (P<0.005). However, with lower concentrations, short‐term exposure did not cause significant changes in ROS level and oxygen consumption rate. We are currently exposing cells to lower concentrations for longer period of time. Conclusions It is generally agreed that oxidative stress plays important roles in acute cadmium poisoning. Interestingly, our data demonstrated decreased oxygen consumption and lower ROS level upon acute high dose cadmium exposure. The possible mechanism could be that during acute response to high dose cadmium, metallothioneins (MTs) are induced and act as a potent sequestration mechanism, thus protecting against oxidative stress.