Ethanolic extract of Puhuang (Pollen Typhae) modulates lipopolysaccharide-induced inflammatory response through inducible nitric oxide synthase / cyclooxygenase-2 signaling in RAW 264.7 macrophage.

OBJECTIVE: To evaluate the immune modulatory response of Puhuang (Pollen Typhae), ethanolic extract of dried pollens (TP-E) and charcoal activated pollens (CTP-E) were used for their phytochemical evaluation and their modulatory response against lipopolysaccharide (LPS) induced inflammatory activity on RAW264.7 macrophage cells. METHODS: Biochemical assays were carried out to quantify the 1,1-Diphenyl-2-picrylhydrazyl Radical Scavenging Activity, Reducing Power, Ferrous ion chelating ability and total polyphenol content and flavonoids. Non-toxic dose of the extract (TP-E and CTP-E) was chosen based on 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Effect of TP-E and CTP-E on lipopolysaccharides-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression was measured by Western blot and quantitative PCR (qRT-PCR). Expression of inflammatory cytokines, such as interleukins (IL-1ß and IL-6) and tumor necrosis factor α (TNF-α), was quantified using qRT-PCR. Mitogen-activated protein kinase pathway was analyzed using Western blot. RESULTS: Phytochemical analysis revealed that both TP-E and CTP-E have strong antioxidant activities and high flavonoid and phenolic contents. TP-E and CTP-E effectively inhibit the expression of iNOS and COX-2, thereby inhibiting its downstream proinflammatory regulators, the extracellular signal-related kinase-1/2, that decreases the expression of IL-1ß, IL-6 and TNF-α. CONCLUSION: Phytochemical constituents present in Typha angustifolia Linn could be used for treating inflammation-related diseases.

A systematic identification of anti-inflammatory active components derived from Mu Dan Pi and their applications in inflammatory bowel disease.

Mu Dan Pi (MDP), also known as Moutan Cortex Radicis, is a traditional Chinese medicine used to treat autoimmune diseases. However, the impact of MDP and its principal active compounds on inflammatory bowel disease (IBD) is uncertain. This study therefore systemically assessed the anti-inflammatory effects of MDP and its known active compounds in IBD. The anti-inflammatory activities of water extract and individual compounds were screened by NF-κB and interferon regulatory factor (IRF) reporter assays in THP-1 cells induced with either Toll-like receptor or retinoic acid inducible gene I/melanoma differentiation-associated gene 5 activators and further verified in bone marrow-derived macrophages. MDP water extract significantly inhibited the activation of NF-κB and IRF reporters, downstream signaling pathways and the production of IL-6 and TNF-α, in a dose-dependent manner. Among 5 known active components identified from MDP (1,2,3,4,6-penta-O-galloyl-ß-d-glucose [PGG], gallic acid, methyl gallate, paeoniflorin, and paeonol), PGG was the most efficient at inhibiting both reporters (with an IC50 of 5-10 µM) and downregulating IL-6 and TNF-α. Both MDP powder for clinical use and MDP water extract, but not PGG, reduced colitis and pathological changes in mice. MDP and its water extract show promise as a novel therapy for IBD patients.