Modulation of PI3K/AKT signaling and DFT modeling via selected pharmaceutical compounds attenuates carrageenan-induced inflammation and oxidative stress in rats.

Abstract

The main goal of the current study is to estimate the in vivo anti-inflammatory/antioxidant ability of four selected pharmaceutical compounds: bisoprolol (Biso), piracetam (Pirc), clopidogrel (Clop), and cinnarizine (Cinna). Indomethacin (Indo) was used as a reference drug to perform a realistic comparison between the four compounds and the Indo in vivo through tracking PI3K/AKT signaling and computational chemistry via density functional theory (DFT) modeling to analyze the electrostatic potential across the molecule and provide insight into the regions for receptor binding of the studied compounds. To achieve the safe dose of these compounds, cytotoxicity was performed against isolated adipose tissue-derived mesenchymal stem cells (ADMSCs) using MTT assay. In vivo determination of anti-inflammatory/antioxidant biochemical and genetic parameters of the tested compounds against rats' paw carrageenan (Carg)-induced inflammation was assessed. The data showed that there was no significant different in cell viability of ADMSCs until dose 10 µg/ml, so we used this concentration for in vivo experiments. Carg high significantly increased the volume of the paw edema at 120 min and 180 min compared to the control group (p < 0.01). Cinna (10 mg/kg), relatively similar to Indo, was the most anti-inflammatory compound among others, followed by Clop and Pirc, where they decreased the volume of the paw edema significantly (p < 0.01) at 120 min and 180 min compared to the Carg-group. Microscopic examination confirmed the above results indicating that paw tissue of Carg-group shows edema formation and massive inflammation compared with control. In comparison to the control group, Carg high significantly increased the malondialdehyde (MDA) levels (p < 0.01), whereas, at the concentration 10 mg/kg of the tested compounds, the MDA concentrations significantly reduced, especially the Clop, Cinna, and Indo-treated groups. On the contrary, total antioxidant capacity (TAC) and 5-lipoxygenase (5-LOP) concentrations were significantly decreased in Carg-group (p < 0.01) compared with control. Cyclooxygenase-2 (COX-2), phosphoinositide 3-kinase (PI3k), and protein kinase B (AKT) gene expressions were high and significantly upregulated in Carg-group compared to control, while the tested compounds downregulated their expressions compared to the Carg-group. Moreover, COX-2, interleukins (IL-10/IL-6/IL-4), PI3k, and AKT protein concentrations were high and significantly increased in Carg-group compared to control, however the tested compounds were high and significantly decreased their concentrations compared to the Carg-group. DFT modeling aligned with the biochemical data and indicated that Cinna emerges as the most reactive drug with high polarizability (302.741 a.u.), a relative small FMOs energy gap (ΔE 5.002 eV), relative low molecular hardness (2.501 eV), relative high softness (0.400 eV^-1), and distinct nucleophilic/electrophilic interaction sites, indicating strong specific interactions with biological receptor. In conclusion, this study revealed the ability of Cinna to potentially suppress inflammation in in vivo Carg-induced rat paw inflammation model through inhibition of PI3K/AKT signaling and DFT modeling mediated by oxidative stress and inflammatory mediators.