eupatilin and Disease-Models--Animal

Psoriasis is a chronic inflammatory skin disease that is currently incurable and causes long-term distress to patients. Therefore, there is an urgent need to develop safe and effective psoriatic drugs. Eupatilin is a natural flavone, that has a variety of pharmacological effects. However, the anti-psoriatic effect of eupatilin and its underlying mechanism remain unclear.. HaCaT cells were treated with 20 μg/mL LPS for 24 h to establish the proliferation model of HaCaT cells. Cell viability was measured by MTT assay. Western blotting was used to detect the expression of p-p38 MAPK, p38 MAPK, p-NF-κB p65 and NF-κB p65 in HaCaT cells. Imiquimod (IMQ) was used to induce psoriasis-like mouse model. Psoriasis Area Severity Index (PASI) score was used to evaluate the degree of skin injury, H&E staining was used to observe the pathological damage of skin tissues, and the expression levels of TNF-α, IL-6, IL-23 and IL-17 in the serum were detected by enzyme-linked immunosorbent assay (ELISA).. Eupatilin could inhibit the hyperproliferation of LPS-stimulated HaCaT cells through p38 MAPK/NF-κB signaling pathway in vitro. In psoriatic mice, eupatilin could significantly reduce skin erythema, scales and thickening scores, ameliorate skin histopathological lesions, and decrease the levels of TNF-α, IL-6, IL-23 and IL-17 in the serum.. Eupatilin had a good anti-proliferative effect in LPS-stimulated HaCaT cells, and significantly alleviated IMQ-induced psoriasis-like lesions in mice. Eupatilin was a promising drug for the treatment of psoriasis.

Topics: Animals; Cell Proliferation; Disease Models, Animal; Imiquimod; Interleukin-17; Interleukin-23; Interleukin-6; Keratinocytes; Lipopolysaccharides; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Psoriasis; Skin; Skin Diseases; Tumor Necrosis Factor-alpha

To investigate the effect of eupatilin in asthma treatment, we evaluated its therapeutic effect and related signal transduction in OVA-induced asthmatic mice and LPS-stimulated RAW264.7 cells. The BALF was tested for changes in lung inflammatory cells. Th2 cytokines in the BALF and OVA-IgE in the serum were measured by ELISA. H&E and PAS staining were used to evaluate histopathological changes in mouse lungs. The key proteins NF-κB, MAPK, and Nrf2 in lung tissues were quantitatively analyzed by Western blotting. Finally, we evaluated the effect of eupatilin on cytokines and related protein expression in LPS-stimulated RAW 264.7 cells in vitro. In OVA-induced asthmatic mice, eupatilin reduced the numbers of inflammatory cells, especially neutrophils and eosinophils. Eupatilin also decreased the levels of IL-5, IL-13 in the BALF and OVA-IgE in the serum. Furthermore, eupatilin inhibited the activation of NF-κB and MAPK pathways and increased the expression of Nrf2 in OVA-induced asthmatic mice. In vitro, eupatilin significantly reduced LPS-stimulated NO, IL-6, and ROS production. Additionally, the NF-κB, MAPK, and Nrf2 protein expression in LPS-stimulated RAW264.7 cells was consistent with that in OVA-induced asthmatic lung tissues. In summary, eupatilin attenuated OVA-induced asthma by regulating NF-κB, MAPK, and Nrf2 signaling pathways. These results suggest the utility of eupatilin as an anti-inflammatory drug for asthma treatment.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Female; Flavonoids; Gene Expression Regulation; Lipopolysaccharides; MAP Kinase Signaling System; Mice; Molecular Structure; Neutrophils; NF-E2-Related Factor 2; NF-kappa B; Ovalbumin; RAW 264.7 Cells; Reactive Oxygen Species

Myofibroblasts are known to play a key role in the development of idiopathic pulmonary fibrosis (IPF). Two drugs, pirfenidone and nintedanib, are the only approved therapeutic options for IPF, but their applications are limited due to their side effects. Thus, curative IPF drugs represent a huge unmet medical need.. A mouse hepatic stellate cell (HSC) line was established that could robustly differentiate into myofibroblasts upon treatment with TGF-β. Eupatilin was assessed in diseased human lung fibroblasts from IPF patients (DHLFs) as well as in human lung epithelial cells (HLECs). The drug's performance was extensively tested in a bleomycin-induced lung fibrosis model (BLM). Global gene expression studies and proteome analysis were performed.. Eupatilin attenuated disease severity of BLM in both preventative and therapeutic studies. The drug inhibited the in vitro transdifferantiation of DHLFs to myofibroblasts upon stimulation with TGF-β. No such induction of the in vitro transdifferantiation was observed in TGF-β treated HLECs. Specific carbons of eupatilin were essential for its anti-fibrotic activity. Eupatilin was capable of dismantling latent TGF-β complex, specifically by eliminating expression of the latent TGF-β binding protein 1 (LTBP1), in ECM upon actin depolymerization. Unlike eupatilin, pirfenidone was unable to block fibrosis of DHLFs or HSCs stimulated with TGF-β. Eupatilin attenuated phosphorylation of Smad3 by TGF-β. Eupatilin induced myofibroblasts to dedifferentiate into intermediate HCS-like cells.. Eupatilin may act directly on pathogenic myofibroblasts, disarming them, whereas the anti-fibrotic effect of pirfenidone may be indirect. Eupatilin could increase the efficacy of IPF treatment to curative levels.

Topics: Animals; Bleomycin; Cell Differentiation; Cell Line; Cell Transdifferentiation; Disease Models, Animal; Extracellular Matrix; Fibroblasts; Flavonoids; Gene Expression Regulation; Hepatic Stellate Cells; Humans; Idiopathic Pulmonary Fibrosis; Latent TGF-beta Binding Proteins; Mice; Myofibroblasts; Phosphorylation; Smad3 Protein; Transforming Growth Factor beta

Despite the higher morbidity of ulcerative colitis (UC), available treatments remain unsatisfactory in recent years. A natural flavone eupatilin (Eup) is known to inhibit the intestinal contraction.. The protective role of Eup in intestinal inflammation remains unclear. This study attempted to determine the bioactivity of Eup against colitis and clarify the mechanism of action.. The in vitro effects of Eup on lipopolysaccharide-induced human THP-M macrophage activation and tumour necrosis factor-α (TNF-α)-damaged intestinal epithelial (NCM460) cells were explored to clarify its potential protective effects. Then, the alleviative efficacy of Eup was established in dextran sodium sulphate (DSS)-induced mice colitis.. Pathological diagnosis, immunohistochemical staining, and reverse transcriptase PCR analysis as well as western blot analysis were employed in the current study.. Eup clearly inhibited inflammatory responses in LPS-stimulated macrophages. Eup also clearly stabilized colonic epithelia by down-regulating overexpression of tight junction proteins and NADPH oxidases 4 (NOX4), and by promoting AMP-activated protein kinase (AMPK) activation in TNF-α-stimulated NCM460 cells. In addition, in vivo study demonstrated that Eup treatment clearly ameliorated the symptoms and pathologic changes of colitis mice. The therapeutic effect of Eup was found to be reduced when compound C (an AMPK pharmacological inhibitor) was given to mice.. The study successfully demonstrated that Eup ameliorated DSS-induced mice colitis by suppressing inflammation and maintaining the integrity of the intestinal epithelial barrier via AMPK activation. The results provide valuable guidance for using Eup in UC treatment.

Topics: AMP-Activated Protein Kinases; Animals; Colitis; Dextran Sulfate; Disease Models, Animal; Epithelial Cells; Female; Flavonoids; Humans; Intestinal Mucosa; Macrophages; Mice; Mice, Inbred C57BL; NADPH Oxidase 4; Signal Transduction; THP-1 Cells; Tumor Necrosis Factor-alpha

Eupatilin is the main active component of DA-9601, an extract from Artemisia. Recently, eupatilin was reported to have anti-inflammatory properties. We investigated the anti-arthritic effect of eupatilin in a murine arthritis model and human rheumatoid synoviocytes. DA-9601 was injected into collagen-induced arthritis (CIA) mice. Arthritis score was regularly evaluated. Mouse monocytes were differentiated into osteoclasts when eupatilin was added simultaneously. Osteoclasts were stained with tartrate-resistant acid phosphatase and then manually counted. Rheumatoid synoviocytes were stimulated with TNF-α and then treated with eupatilin, and the levels of IL-6 and IL-1β mRNA expression in synoviocytes were measured by RT-PCR. Intraperitoneal injection of DA-9601 reduced arthritis scores in CIA mice. TNF-α treatment of synoviocytes increased the expression of IL-6 and IL-1β mRNAs, which was inhibited by eupatilin. Eupatilin decreased the number of osteoclasts in a concentration dependent manner. These findings, showing that eupatilin and DA-9601 inhibited the expression of inflammatory cytokines and the differentiation of osteoclasts, suggest that eupatilin and DA-9601 is a candidate anti-inflammatory agent.

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Differentiation; Cells, Cultured; Collagen Type II; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Female; Flavonoids; Humans; Inflammation; Interleukin-1beta; Interleukin-6; Lymph Nodes; Mice; Mice, Inbred DBA; Monocytes; Osteoclasts; Plant Extracts; RNA, Messenger; Synovial Membrane; T-Lymphocytes, Regulatory; Tumor Necrosis Factor-alpha

The transplantation of isolated pancreatic islets is a promising treatment for diabetes. 5,7-dihydroxy-3,4,6-trimethoxyflavone (Eupatilin), a pharmacologically active flavone derived from the Artemisia plant species, has been reported to have antioxidant and anti-inflammatory activities. This study examines the hypothesis that preoperative eupatilin treatment can attenuate ischemic damage and apoptosis before islet transplantation.. Islets isolated from Balb/c mice were randomly divided into 2 groups, and cultured in medium supplemented with or without eupatilin. In vitro islet viability and function were assessed. After treatment with a cytokine cocktail consisting of tumor necrosis factor (TNF)-α, interferon (INF)-γ, and interleukin (IL)-1β, islet cell viability, function, and apoptotic status were determined. The glutathione (GSH) and nitrous oxide (NO) levels were also measured. Proteins related to apoptosis were analyzed using Western blotting.. There was no difference in cell viability between the 2 groups. Islets cultured in the medium supplemented with eupatilin showed 1.4-fold higher glucose-induced insulin secretion than the islets cultured in the medium without eupatilin. After treatment with a cytokine cocktail, glucose-induced insulin release and the total insulin content of the islets were significantly improved in eupatilin-pretreated islets compared with islets not treated with eupatilin. Apoptosis was significantly decreased, and GSH levels were elevated in the eupatilin-pretreated group. Cytokine-only treated islets produced significantly higher levels of NO, iNOS, and caspase-3 than islets pretreated with eupatilin before cytokine treatment.. These results suggest that preoperative eupatilin administration enhances islet function before transplantation and attenuates the cytokine-induced damage associated with NO production and apoptosis.

Topics: Animals; Apoptosis; Cell Survival; Disease Models, Animal; Drugs, Chinese Herbal; Female; Flavonoids; Islets of Langerhans; Islets of Langerhans Transplantation; Mice; Mice, Inbred BALB C; Reperfusion Injury