Calcarea carbonica treatment rescues lipopolysaccharide-induced inflammatory response in human mononuclear cells via downregulation of inducible cyclooxygenase pathway

• 作者: Anil Khurana Debadatta Nayak Madan L.B.Bhatt Raj K.Manchanda Shailendra K.Saxena Srinivasulu Gadugu Swatantra Kumar Vimal K.Maurya
• 来源期刊: 结合医学学报（英文版）
• 年，卷（期）: 2020,（5）
• 所属期刊栏目: Original Research Artic1e (Experiment)
• 页码范围: 441-449
• 页数: 9页
• 语种: 英文

摘要

Objective: Prolonged use of nonsteroidal anti-inflammatory drugs is associated with severe side effects and toxicity.Therefore,we studied the anti-inflammatory role of Calcarea carbonica which had minimal toxicity at the low doses.Methods: THP-1 human mononuclear cells were treated with C.carbonica to evaluate the 50％ cytotoxicity concentration (CC50) and 50％ effective concentration (EC50).Cell survival was evaluated in lipopolysaccharide-stimulated C.carbonica-treated cells.Nitric oxide (NO) and tumor necrosis factor-α (TNF-α) were measured to evaluate the anti-inflammatory activity of C.carbonica.Cyclooxygenase-2 (COX-2) protein expression was determined by Western blotting analysis,and the interaction of C.carbonica with the COX-2 protein was evaluated using molecular docking simulation.Results: The CC50 and EC50 of C.carbonica were found to be 43.26 and 11.99 μg/mL,respectively.The cell survival assay showed a 1.192-fold (P =0.0129),1.443-fold (P =0.0009) and 1.605-fold (P =0.0004) increase in cell survival at 24,48 and 72 h after initiating C.carbonica treatment,respectively.C.carbonica-treated cells showed a reduction in NO levels by 2.355 folds (P =0.0001),2.181 folds (P =0.0001) and 2.071 folds (P =0.0001) at 24,48 and 72 h,respectively.The treated cells also showed a reduction in TNF-α levels by 1.395 folds (P =0.0013),1.541 folds (P =0.0005) and 1.550 folds (P =0.0005) at 24,48 and 72 h,respectively.In addition,a 1.193-fold reduction (P =0.0126) in COX-2 protein expression was found in C.carbonica-treated cells.The molecular docking showed interaction of C.carbonica with the phenylalanine 367 residue present in active site of Cox-2.Conclusion: C.carbonica exhibited anti-inflammatory properties in lipopolysaccharide-stimulated cells by significantly reducing NO production and TNF-α level through downregulation of the COX-2 protein.This effect is probably mediated through interaction of C.carbonica with the phenylalanine 367 residue present in active site of Cox-2.