In vitro Evaluation of the Anti-inflammatory Effects of Thymoquinone in Osteoarthritis and in silico Analysis of Inter-Related Pathways in Age-Related Degenerative Diseases

Abstract

Chronic inflammation is a common underlying factor in osteoarthritis (OA) and most age-related degenerative diseases. As conventional therapies help only in partial alleviation of symptoms in OA, stem cell-based therapies and herbal supplements are being widely explored. Thymoquinone (TQ), an active ingredient of Nigella sativa is reported to have immunomodulatory, anti-inflammatory and antioxidant properties. We evaluated the effects of TQ on bone marrow MSCs (BM-MSCs) derived from OA patients and its interrelated pathways in inflammation and age-related degenerative diseases using Ingenuity Pathway Analysis (IPA) as well as possible molecular targets using SwissTargetPrediction. BM-MSCs were derived from OA patients and their stemness properties were characterized by studying the MSCs related CD surface marker expression and differentiation into adipocytes, osteoblasts, and chondrocytes. Treatment with TQ (100 nM-5 µM) demonstrated cell death, especially at higher concentrations. MTT assay demonstrated a significant concentration-dependent decrease in cell viability which ranged from 20.04% to 69.76% with higher doses (300 nM, 1 µM, and 5 µM), especially at 48h and 72h. Additional cell viability testing with CellTiter-Blue also demonstrated a significant concentration-dependent decrease in cell viability which ranged from 27.80 to 73.67% with higher doses (300 nM, 1 µM, 3 µM, and 5 µM). Gene expression analysis following treatment of BM-MSCs with TQ (1 and 3 µM) for 48h showed upregulation of the anti-inflammatory genes IL-4 and IL-10. In contrast, the pro-inflammatory genes namely IFN-γ, TNF-α, COX-2, IL-6, IL-8, IL-16, and IL-12A although were upregulated, compared to the lower concentration of TQ (1 µM) they were all decreased at 3 µM. The pro-apoptotic BAX gene was

Decoding the Role of Astrocytes in the Entorhinal Cortex in Alzheimer’s Disease Using High-Dimensional Single-Nucleus RNA Sequencing Data and Next-Generation Knowledge Discovery Methodologies: Focus on Drugs and Natural Product Remedies for Dementia

“…We have previously shown that natural products such as albiziasaponin-A, iso-orientin, and salvadorin can ameliorate the pathologies associated with AD in vivo ( Rasool et al, 2018 ) and that the natural products could be useful for the treatment of age-related degenerative diseases ( Kalamegam et al, 2020 ). In addition, we have recently shown that NGKD platforms can be successfully used to find drugs and natural products that may reverse disease-specific gene signatures ( Pushparaj et al, 2021 ).…”

Thymoquinone Attenuates Immune Mediated Liver Injury Induced by Concanavalin A in Mice

“…Studies showed that thymoquinone has immunomodulatory and anti-inflammatory effects, that prevent the biosynthesis of important mediators in inflammatory processes and reduce proinflammatory cytokines such as interleukins (ILs) and TNF α. Besides, thymoquinone showed the immunomodulatory role in cellular and humoral immunity (19) (20) . Analysis of data obtained from the current study revealed that concanavalin A administration by retro-orbital route result in a remarkable elevation in both AST and ALT serum activities in comparison to the negative control group which received normal saline through retroorbital rout.…”

Antiatherogenic Effects of Quercetin in the THP-1 Macrophage Model In Vitro, With Insights Into Its Signaling Mechanisms Using In Silico Analysis

“…In reverse screening, the combined score helps to calculate the probability of targeting a given protein. Dual-based reverse screening has demonstrated high performance in predicting macromolecular targets (Kalamegam et al, 2020).…”