Zinc Modulates the Response to Apoptosis in an In Vitro Model with High Glucose and Inflammatory Stimuli in C2C12 Cells
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Zinc Modulates the Response to Apoptosis in an In Vitro Model with High Glucose and Inflammatory Stimuli in C2C12 Cells Mónica Andrews-Guzmán 1 & Manuel Ruz 2 & Miguel Arredondo-Olguín 1 Received: 9 April 2020 / Accepted: 17 August 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Apoptosis is programmed cell death and its alteration is related to cancer, neurologic, autoimmune, and chronic diseases. A number of factors can affect this process. The aim of this paper is to study the effect of supplemental zinc on apoptosis-related genes in C2C12 myoblast cells after being challenged with a series of stimuli, such as high glucose, insulin, and an inflammatory agent. C2C12 myoblast cells were cultured for 24 h with zinc (Zn) (ZnSO4) 10 or 100 μM and/or glucose 10 or 30 mM. In addition to these stimuli, the cells were challenged with insulin 1 nM or interleukin-6 (IL-6) 5 nM. The mRNA expression of proapoptotic genes caspase 3 and Fas, the antiapoptotic genes, Xiap and Bcl-xL and the ratio of pro-/antiapoptotic genes Bax/Bcl2, were determined by qRT-PCR. The expression of caspase-3 gene was significantly increased in the presence of the combination high Zn/high glucose with and without the presence of insulin and IL6 in the culture medium Fas expression instead, showed uneven responses. The expression of Bcl-xL and Xiap was increased in most conditions by having high Zn in the medium regardless of the presence of insulin or IL6. Bax/Bcl2 ratio was decreased in the presence of high Zn. Zn was able to stimulate the expression of antiapoptotic genes. This effect was specially noted in high-glucose conditions with and without the presence of insulin. This effect is partially overridden by the presence of an inflammatory agent such as IL-6. Keywords Zinc . Apoptosis . Bax/Bcl2 ratio . Caspase-3 . Fas
Abbreviations DMT1 Divalent metal transporter 1 ZnT Zinc transporter FBS Fetal bovine serum qRT-PCR Quantitative reverse transcription– polymerase chain reaction B2M Beta-2 microglobulin ANOVA Analysis of variance SEM Standard error of the mean
* Miguel Arredondo-Olguín [email protected] 1
Micronutrient Laboratory, Institute of Nutrition and Food Technology, University of Chile, El Líbano 5524, Macul, Santiago, Chile
2
Department of Nutrition, Faculty of Medicine, University of Chile, Avenida Independencia 1027, Independencia, Santiago, Chile
Introduction There is a series of conditions that can lead to inflammation and oxidative stress, such as high circulating glucose [1] and saturated fatty acids [2]. These, along with iron [3] or copper [4], could trigger a number of additional adverse effects. In addition, decreased insulin production in the pancreas and increased inflammation in liver and muscle are able to induce alterations in glucose uptake, glucose output, and increased apoptosis in these tissues [5]. Zinc (Zn) is an essential micronutrient that plays a role in processes like DNA/RNA synthesis, cell division, and apoptosis [6]. Zn deficiency has been related to chronic dis
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