Gadodiamide Induced Autophagy and Apoptosis in Human Keratinocytes

Abstract

Background/Aim: Gadolinium has been reported to cause liver lobular necrosis and nephrogenic systemic fibrosis. However, its toxicity to the skin remains unknown. This study aimed to investigate the effect of a high dose of gadolinium-based contrast agent gadodiamide on the human keratinocyte HaCaT cell line. Materials and Methods: Cell viability was assessed using MTT assay, and autophagy was assessed using acridine orange and LysoTracker Red staining. Western blotting was performed to verify the changes in Bcl2 and Bax levels. Results: The viability of HaCaT cells was significantly suppressed after gadodiamide treatment. Interestingly, gadodiamide caused autophagic vacuoles, whereas the autophagy inhibitors 3-methyladenine and chloroquine significantly alleviated autophagic cell death. Simultaneously, gadodiamide induced apoptosis, which was reduced by caspase inhibitors. Gadodiamide also inhibited Bcl-2 expression and promoted Bax expression. Conclusion: Gadodiamide induced both autophagy and apoptosis in HaCaT cells. Physicians should carefully assess the gadodiamide dosage used clinically.Magnetic resonance imaging (MRI) is a contemporary technology that produces three-dimensional images of the anatomical human body and helps in the detection and diagnosis of diseases. The density of contrast agents produces a strong contrast effect; however, patients with severe renal failure may be at risk of a serious illness called nephrogenic systemic fibrosis (NSF) that may be linked to the use of certain gadolinium-based contrast agents (GBCAs) such as gadodiamide (1, 2). GBCAs agents can be applied to the human body, providing anatomical imaging of the head, spinal cord, and vascular artery for doctors to determine the condition of the entire body (3). However, its toxicity to normal skin cells is critical, but has never been investigated.NSF is a serious, late adverse reaction to gadodiamide (4, 5). Thus, the Taiwan Food and Drug Administration suggests the use of the lowest effective dose to prevent possible adverse effects on any part of the human body (6, 7). Longterm ionized gadolinium accumulation may cause fibrosis in various tissues of the human body, including the skin. Early alterations and symptoms of NSF may include pain, itching, swelling, and erythema, which usually start in the legs. Late lesions may include thickening of the skin and subcutaneous tissues, visceral fibrosis, and finally severe symptoms like limb contracture, cachexia, and death (8, 9). However, the detailed mechanisms remain largely unknown. The main purpose of the current study was to explore whether high 603 This article is freely accessible online.

Protective Effects of Jujubosides on 6-OHDA-Induced Neurotoxicity in SH-SY5Y and SK-N-SH Cells

“…Cell viability was assessed using a tetrazolium 3-(4,5-dimethylthiazole-2-yl)-2,5diphenyltetrazolium bromide (MTT) assay as previously described [46,47]. Briefly, the cells were cultured in 96-well plates at a density of 3 × 10 4 cells/well, grown for another day and then exposed to 25 or 50 µM of 6-OHDA, with or without treatment with jujubosides at different concentrations for 1 h prior to 6-OHDA exposure.…”

Metformin induces autophagy of cisplatin-resistant human gastric cancer cells in addition to apoptosis

“…The acidic vesicular organelles (AVOs) formation of happens during cell autophagy. The cisplatin-resistant GC cells (2.5 × 10 5 cells/well) were cultured in 24-well plates and exposed to Metformin (25 and 50 mM) for 48 h. Subsequently, cells were stained with 1 μg/ml Acridine orange (AO) for 30 min at 37 °C, Fluorescent microscope (Leica, Wetzlar, Germany) at 20 × magnification was used to visualize acidic vesicular organelles (AVOs), and analysis on the NucleoCounter NC-3000 using the built-in acridine orange (AO)/acidic vesicular organelles (AVOs) assay program as previously described [ 18 , 19 ].…”

Toxicity Mechanisms of Gadolinium and Gadolinium-Based Contrast Agents—A Review

“…According to the gathered data, several signaling pathways have been implicated in Gd (III) mechanisms of toxicity, such as MAPK/ERK (mitogen-activated protein kinase/extracellular signal-regulated kinase), PI3K/Akt (phosphoinositide-3-kinase/protein kinase B), and EGFR (epidermal growth factor receptor) signaling [26,54,55,71,84,87,89,90,93], suggesting that Gd (III) interferes with the transduction of molecules involved in the regulation of inflammatory processes, and in cell metabolism, proliferation, growth, and survival. Upregulation of inflammation, oxidative stress, and apoptosis were highlighted as potential mechanisms of Gd (III) cytotoxicity [25,30,34,[38][39][40]42,[60][61][62]64,66,70,76,84,86,91,99,101,102,111,113]. It has been reported that exposure to Gd (III) or GBCAs may induce the expression of several profibrotic chemokines and cytokines, and alter cell growth [11,41,54,59,68,69,72,77,88,89,95,…”

“…Besides lipid peroxidation and ROS production, Gd (III) prompted the formation of autophagic vesicles, also revealing apoptotic and necrotic potential [42,110], pointing towards a multitude of cell death pathways being activated. Indeed, a decrease in cell viability, an increase in cell death through apoptosis, and autophagic activation have been associated with Gd (III) toxicity [10,30,34,40,[46][47][48]51,58,60,64,70,99,113]. Mitochondrial dysfunction [38,42,51,91,108,109] and suppressing mitochondria membrane potential [62,101] were also described.…”