N–O Reduction and ROS-Mediated AKT/FOXO1 and AKT/P53 Pathways Are Involved in Growth Promotion and Cytotoxicity of Cyadox

Abstract

Cyadox is a novel derivative of quinoxaline-1,4-dioxides (QdNOs) with the potential to be developed as a feed additive. However, the pharmacological and toxicological bioactive molecules of cyadox and the molecular mechanism of its pharmacological and toxic actions remain unclear. In the present study, cyadox and its main metabolites of cy1, cy4, cy6, and cy12 were selected; the growth promotion characteristic was indicated by the mRNA level of EGF; and the cytotoxicity of cyadox was determined by methylthiazol tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release, and Annexin V-FITC/PI apoptosis detection kit with flow cytometry. The intracellular ROS, cyclin D1, and Akt/P53/FOXO1 signaling pathway were also investigated. Our data suggested that cyadox showed relatively higher activity than its metabolites, and the ROS was generated from N–O reduction of cyadox. Moreover, cyadox (2 μM) activated the Akt and increased the EGF, cyclin D1, and FOXO1 expression levels. Cyadox (100 μM) induced cytotoxicity in L02 cells in a concentration- and time-dependent manner. Additionally, the activated P53 pathway, hyperactivated Akt, and apoptosis were found in L02 cells after incubation with 100 μM cyadox. Our data demonstrated that Akt promoted cell survival when it was mildly activated by cyadox at 2 μM, and Akt leads to apoptosis when it was severely activated by cyadox at 100 μM. Thus, the present study revealed that N–O reduction of cyadox and ROS-mediated AKT/FOXO1 and AKT/P53 pathways were involved in growth promotion and cytotoxicity of cyadox.

High cholesterol induces apoptosis and autophagy through the ROS-activated AKT/FOXO1 pathway in tendon-derived stem cells

“…We have already found that cholesterol upregulates ROS levels in TDSCs [11]. ROS affect a variety of signaling pathways, including the AKT/ FOXO1 signal pathway [19]. The activation of AKT promotes cell growth and proliferation and increases cell resistance to apoptosis in many cell types [20].…”

The role of cyadox and recombinant growth hormone (rGH) on the promotion of growth through epigenetics

“…Currently, quite many studies have suggested that cyadox can improve the growth performance of animals including pigs and poultry [6,7,[37][38][39][40]. Antimicrobials including cyadox might in uence growth via an endocrine axis such as IGF-1, GH and epidermal growth factor (EGF) [41].…”

“…Quinoxaline, 1,4-dioxide derivatives (QdNOs), consisting of one or two acyclic chain moieties combined with a quinoxaline ring, are antibacterial and growth promoting agents and have a wide range of biological properties, including antibacterial, antitubercular, anticancer, anti-candida, and antiprotozoal [1][2][3][4][5][6][7]. Previous reports suggest that these drugs can alter intestinal micro ora, improve protein utilization, and increase protein synthesis in vivo [1].…”

Cyadox regulates the transcription of different genes by activation of the PI3K signaling pathway in porcine primary hepatocytes

“…TK is required for cancer growth because of its ability to affect the production of NAPDH to counteract oxidative stress . Our findings demonstrated that the reactive oxygen species (ROS) was generated from N‐O reduction of cyadox in L02 cells (normal human liver cells) and cyadox upregulates EGF expression by the ROS and activating Akt signaling . We inferred that ROS from the metabolism of cyadox might also promote the TK expression besides EGF and both were involved in growth promotion of cyadox.…”