Calcarea carbonica treatment rescues lipopolysaccharide-induced inflammatory response in human mononuclear cells via downregulation of inducible cyclooxygenase pathway.
10.1016/j.joim.2020.06.001
- Author:
Swatantra KUMAR
1
;
Vimal K MAURYA
1
;
Debadatta NAYAK
2
;
Anil KHURANA
2
;
Raj K MANCHANDA
2
;
Srinivasulu GADUGU
3
;
Madan L B BHATT
1
;
Shailendra K SAXENA
4
Author Information
1. Centre for Advanced Research, Faculty of Medicine, King George's Medical University, Lucknow 226003, India.
2. Central Council for Research in Homoeopathy, Ministry of Ayush, Janakpuri, New Delhi 110058, India.
3. Department of Medicine, Jaisoorya and Potti Sreeramulu Government Medical College, Hyderabad 500013, India.
4. Centre for Advanced Research, Faculty of Medicine, King George's Medical University, Lucknow 226003, India. Electronic address: shailen@kgmcindia.edu.
- Publication Type:Journal Article
- Keywords:
Anti-inflammatory;
Calcarea carbonica;
Cyclooxygenase-2;
Inflammation;
Nitric oxide;
Tumor necrosis factor-α
- From:
Journal of Integrative Medicine
2020;18(5):441-449
- CountryChina
- Language:English
-
Abstract:
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) and 50% effective concentration (EC). 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 CC and EC 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.
