Anti-oxidant and Anti-inflammatory Effects of Ethanol Extract from Polygala sibirica L. var megalopha Fr. on Lipopolysaccharide-Stimulated RAW264.7 Cells.
10.1007/s11655-023-3602-7
- Author:
Cheng-Liu YANG
1
;
Shi-Bo WANG
1
;
Wen-Ping HE
1
;
Jin-Juan LIU
2
Author Information
1. Key Laboratory of Biotechnology for Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou, Jiangsu Province, 221116, China.
2. Key Laboratory of Biotechnology for Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou, Jiangsu Province, 221116, China. jjlbest@jsnu.edu.cn.
- Publication Type:Journal Article
- Keywords:
Chinese medicine;
Polygala sibirica L. var megalopha Fr.;
RAW264.7 cells;
anti-inflammatory;
anti-oxidant;
lipopolysaccharide
- MeSH:
Animals;
Mice;
Antioxidants/pharmacology*;
Lipopolysaccharides/pharmacology*;
Polygala;
Transcription Factor RelA/metabolism*;
Tumor Necrosis Factor-alpha/metabolism*;
Ethanol/chemistry*;
Interleukin-6/metabolism*;
Anti-Inflammatory Agents/chemistry*;
Reactive Oxygen Species/metabolism*;
Cyclooxygenase 2/metabolism*;
Nitrites/metabolism*;
NF-kappa B/metabolism*;
Nitric Oxide/metabolism*;
Superoxide Dismutase/metabolism*;
RNA, Messenger;
Nitric Oxide Synthase Type II/metabolism*
- From:
Chinese journal of integrative medicine
2023;29(10):905-913
- CountryChina
- Language:English
-
Abstract:
OBJECTIVE:To investigate the anti-oxidant and anti-inflammatory effects of ethanol extract of Polygala sibirica L. var megalopha Fr. (EEP) on RAW264.7 mouse macrophages.
METHODS:RAW264.7 cells were pretreated with 0-200 µg/mL EEP or vehicle for 2 h prior to exposure to 1 µg/mL lipopolysaccharide (LPS) for 24 h. Nitric oxide (NO) and prostaglandin (PGE2) production were determined by Griess reagent and enzyme-linked immunosorbent assay (ELISA), respectively. The mRNA levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor α (TNF-α), interleukin-1beta (IL-1β), and IL-6 were determined using reverse transcription polymerase chain reaction (RT-PCR). Western blot assay was used to determine the protein expressions of iNOS, COX-2, phosphorylation of extracellular regulated protein kinases (ERK1/2), c-Jun N-terminal kinase (JNK), inhibitory subunit of nuclear factor Kappa B alpha (Iκ B-α) and p38. Immunofluorescence was used to observe the nuclear expression of nuclear factor-κ B p65 (NF-κ B p65). Additionally, the anti-oxidant potential of EEP was evaluated by reactive oxygen species (ROS) production and the activities of catalase (CAT) and superoxide dismutase (SOD). The 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH), superoxide anion (O2-) radical and nitrite scavenging activity were also measured.
RESULTS:The total polyphenol and flavonoid contents of EEP were 23.50±2.16 mg gallic acid equivalent/100 g and 43.78±3.81 mg rutin equivalent/100 g. With EEP treatment (100 and 150 µg/mL), there was a notable decrease in NO and PGE2 production induced by LPS in RAW264.7 cells by downregulation of iNOS and COX-2 mRNA and protein expressions (P<0.01 or P<0.05). Furthermore, with EEP treatment (150 µg/mL), there was a decrease in the mRNA expression levels of TNF-α, IL-1β and IL-6, as well as in the phosphorylation of ERK, JNK and p38 mitogen-activated protein kinase (MAPK, P<0.01 or P<0.05), by blocking the nuclear translocation of NF-κ B p65 in LPS-stimulated cells. In addition, EEP (100 and 150 µg/mL) led to an increase in the anti-oxidant enzymes activity of SOD and CAT, with a concomitant decrease in ROS production (P<0.01 or P<0.05). EEP also indicated the DPPH, OH, O2- radical and nitrite scavenging activity.
CONCLUSION:EEP inhibited inflammatory responses in activated macrophages through blocking MAPK/NF-κ B pathway and protected against oxidative stress.
