Prim-O-glucosylcimifugin attenuates lipopolysaccharide- induced inflammatory response in RAW 264.7 macrophages
Jie Zhou1, Yuan-Yuan Sun2, Meng-Yao Sun3, Wei-An Mao4, Li Wang1, Jian Zhang1, Hong Zhang5
1 Department of Dermatology, Seventh People's Hospital of Shanghai University of TCM, Shanghai 200137, China
2 Department of Clinical Medicine, Bengbu Medical College, Anhui 233000, China
3 Department of Pharmaceutical Botany, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
4 Xinchang Community Health Service Center, Pudong New Area, Shanghai 201314, China
5 Department of Pharmaceutical Botany, School of Pharmacy, Second Military Medical University, Shanghai 200433; Central Laboratory, Seventh People's Hospital of Shanghai University of TCM, Shanghai 200137, China
Central Laboratory, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, 358 Datong Road, Shanghai 200137
Xinchang Community Health Service Center, Pudong New Area, 58 West Pailou Road, Shanghai 201314
Source of Support: None, Conflict of Interest: None
Background: Radix Saposhnikoviae (RS) exerts anti-inflammatory, analgesic, antipyretic, antioxidation effects and has been used in traditional Chinese medicine to treat common colds, headache, and rheumatoid arthritis. Prim-O-glucosylcimifugin (POG) is the highest content chromone and one of the major active constituents in RS. Objective: The study was aimed to explore the anti-inflammation effects of POG in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Materials and Methods: Cell viability was detected by Cell Counting Kit-8 assay. Production of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 was assessed by enzyme-linked immunosorbent assay. Real-time polymerase chain reaction and Western blot were performed to analyze mRNA and protein levels, respectively. Results: During the whole experiment, 15, 50, and 100 μg/mL of POG had no cytotoxicity on RAW 264.7 cells. POG dose-dependently inhibited the production of NO, TNF-α, IL-1β, and IL-6 that were induced by LPS. POG treatment downregulated the mRNA and protein expression inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2) in LPS-activated RAW 264.7 macrophages in a concentration-dependent manner. Furthermore, LPS-induced JAK2/STAT3 activation was prevented in RAW 264.7 macrophages by POG treatment. STAT3 overexpression significantly reversed the effects of POG on LPS-activated RAW 264.7 macrophages. Conclusion: These results demonstrate that POG exerts anti-inflammatory effects through the inhibition of iNOS and COX-2 expression by inhibiting the phosphorylation of JAK2/STAT3.
Abbreviations used: LPS: Lipopolyssacharide; NO: Nitric oxide; TNF-α: Tumor necrosis factor-α; IL: Interleukin; RS: Radix Saposhnikoviae; POG: Prim-O-glucosylcimifugin; iNOS: Inducible NO synthase; COX2: Cyclooxygenase; FBS: Fetal bovine serum; DMSO: Dimethylsulfoxide; CCK-8: Cell Counting Kit; RIPA: Radio immunoprecipitation assay buffer; ECL: Enhanced chemiluminescence; SD: Standard deviation; ELISA: Enzyme-Linked immunosorbent assay.
Dr. Hong Zhang,