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 a...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(CC_(50)) and 50% effective concentration(EC_(50)). Cell survival was evaluated in lipopolysaccharide-stimulated C. carbonica-treated cells. Nitric oxide(NO) and tumor necrosis factor-a(TNF-a) 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 CC_(50) and EC_(50) of C. carbonica were found to be 43.26 and 11.99 mg/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 TNFa levels by 1.395 folds(P = 0.0013), 1.541 folds(P = 0.0005) and 1.550 folds(P = 0.0005) at 24, 48 and72 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-a 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.展开更多
基金supported by Central Council for Research in Homoeopathy,Ministry of Ayush,Government of India。
文摘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(CC_(50)) and 50% effective concentration(EC_(50)). Cell survival was evaluated in lipopolysaccharide-stimulated C. carbonica-treated cells. Nitric oxide(NO) and tumor necrosis factor-a(TNF-a) 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 CC_(50) and EC_(50) of C. carbonica were found to be 43.26 and 11.99 mg/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 TNFa levels by 1.395 folds(P = 0.0013), 1.541 folds(P = 0.0005) and 1.550 folds(P = 0.0005) at 24, 48 and72 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-a 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.
