wedelolactone and Inflammation

Cholestatic liver diseases (CLD) comprise a variety of disorders of bile formation, which causes chronic exposure to bile acid (BA) in the liver generally and results in hepatotoxicity and progressive hepatobiliary injury. Wedelolactone (7-methoxy-5, 11, 12-trihydroxy-coumestan, WED), the natural active compound derived from Ecliptae Herba, has been reported with valuable bioactivity for liver protection. Nevertheless, the effect of WED on cholestatic liver injury (CLI) remains unexplored.. The present study aims to elucidate the protective effect of WED on Alpha-naphthylisothiocyanate (ANIT)-induced CLI mice, and to investigate its potential pharmacological mechanism.. The anit-cholestatic and hepatoprotective effects of WED were evaluated in ANIT-induced CLI mice. Non-targeted metabolomics study combined with ingenuity pathway analysis (IPA) was used to explore the key mechanism of WED. The BA metabolic profile in enterohepatic circulation was analyzed to evaluate the effect of WED in regulating BA metabolism. Furthermore, molecular dynamics (MD) simulation and cellular thermal shift assay (CETSA) were used to simulate and verify the targeting activation of WED on the Farnesoid X receptor (FXR). The core role of FXR in WED promoting BA transportation, and alleviating BA accumulation-induced hepatotoxicity was further evaluated in WT and FXR knockout mice or hepatocytes.. WED dose-dependently alleviated ANIT-induced cholestasis and liver injury in mice, and simultaneously suppressed the signaling pathway of nuclear factor-kappa B/nuclear factor-erythroid 2-related factor 2 (NF-κB/NRF2) to relieve inflammation and oxidative stress. At the metabolite level, WED improved the metabolic disorder in CLI mice focusing on the metabolism of BA, arachidonic acid, and glycerophospholipid, that closely related to the process of BA regulation, inflammation, and oxidative damage. WED targeting activated FXR, which then transcribed its target genes, including the bile salt export pump (BSEP) and the BA transporter, and subsequently increased BA transportation to restore the damaged enterohepatic circulation of BA. Meanwhile, WED alleviated hepatic BA accumulation and protected the liver from BA-induced damage via NF-κB/NRF2 signaling pathway. Furthermore, FXR deficiency suppressed the protective effect of WED in vitro and in vivo.. WED regulated BA metabolism and alleviated hepatic damage in cholestasis. It protected the liver according to adjusted BA transportation and relieved BA accumulation-related hepatotoxicity via FXR-bile acid-NF-κB/NRF2 axis. Our study provides novel insights that WED might be a promising strategy for cholestatic liver disease.

Topics: Animals; Bile Acids and Salts; Chemical and Drug Induced Liver Injury; Cholestasis; Inflammation; Liver; Mice; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress

Rheumatoid arthritis (RA) is an autoimmune disorder associated with joint damage and attendant cardiovascular complications. Wedelolactone (Wed), derived from Eclipta alba, possesses anti-inflammatory activity. Whether Wed regulates RA inflammation and related heart damage remains unknown.. A murine model of collagen-induced arthritis (CIA) was well-established by two subcutaneous injections of type II collagen (days 0 and 21). Wed was then administered via intraperitoneal injection every other day from day 28 to day 48. Joint swelling was monitored and paw thickness was calculated. Histopathological changes in synovial tissues or ankle cartilage were evaluated by hematoxylin and eosin (H&E) and Safranin O-Fast Green staining. The concentrations of inflammatory factors in serum and synovial tissues were detected by ELISA. The qRT-PCR, Western blotting, immunohistochemistry (IHC), and immunofluorescence (IF) were performed to assess receptor activator of nuclear factor kappa ligand (RANKL), matrix metalloprotease (MMP)-3, NLRP3, caspase-1 (pro- and cleaved forms), p-p65, IκBα, p-IκBα, p65, αsmooth-actin 2 (ACTA2), collagen type I and E-cadherin expression. H&E and Masson staining were used to assess the pathological alterations in the heart.. Treatment with Wed ameliorated ankle joint swelling and cartilage degradation. Wed decreased the infiltration of inflammatory cells, the release of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, and IL-18), and the expression of RANKL and MMP-3 in serum and synovial tissues of CIA mice. Moreover, Wed increased the expression of NLRP3 and cleaved-caspase-1 in the synovium, leading to IL-1β and IL-18 secretion. Nuclear factor-kappaB (NF-κB) activation in synovial tissues was suppressed by Wed, as manifested by reduced phosphorylation of p65 and IκBα and nuclear translocation of p65. Furthermore, Wed reduced in CIA mice heart weight/body weight ratio and dampened cardiac inflammation and fibrosis that was accompanied at the mRNA level by down-regulation of ACTA2 and collagen I and up-regulation of E-cadherin.. These findings suggested that Wed attenuated synovial inflammation and joint damage in a mouse model of RA via inhibiting NF-κB/NLRP3 inflammasome activation, and ameliorated RA-induced cardiac complications.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Caspases; Disease Models, Animal; Inflammasomes; Inflammation; Interleukin-18; Mice; NF-kappa B; NF-KappaB Inhibitor alpha; NLR Family, Pyrin Domain-Containing 3 Protein; Synovial Membrane

Wedelolactone exhibits regulatory effects on some inflammatory diseases. However, the anti-inflammatory mechanism of wedelolactone has not been entirely unravelled. Therefore, the present study focuses on investigating the mechanism of wedelolactone on NLRP3 inflammasome in macrophages and its influence on MSU-induced inflammation.. BMDM, J774A.1 and PMA-differentiated THP-1 macrophages were primed with LPS and then stimulated with ATP or nigericin or MSU crystal in the presence or absence of wedelolactone. The cell lysates and supernatants were collected to detect NLRP3 inflammasome components such as NLRP3, ASC and caspase 1, as well as pyroptosis and IL-1β production. In addition, the anti-inflammatory effects of wedelolactone on MSU-induced peritonitis and arthritis mice were also evaluated.. We found that wedelolactone broadly inhibited NLRP3 inflammasome activation and pyroptosis and IL-1β secretion. Wedelolactone also block ASC oligomerization and speck formation. The inhibitory effects of wedelolactone were abrogated by PKA inhibitor H89, which also attenuated wedelolactone-enhanced Ser/Thr phosphorylation of NLRP3 at PKA-specific sites. Importantly, wedelolactone could abate MSU-induced IL-1β production and neutrophils migration into peritoneal cavity, and reduced caspase 1 (p20) and IL-1β expression in the joint tissue of MSU-induced arthritis.. Our results indicate that wedelolactone promotes the Ser/Thr phosphorylation of NLRP3 to inhibit inflammasome activation and pyroptosis partly through potentiating PKA signalling, thus identifying its potential use for treating MSU-induced peritonitis and gouty arthritis.

Topics: Animals; Anti-Inflammatory Agents; Cell Line; Coumarins; Cyclic AMP-Dependent Protein Kinases; Humans; Inflammasomes; Inflammation; Macrophages; Male; Mice, Inbred C57BL; NLR Family, Pyrin Domain-Containing 3 Protein; Phosphorylation; Pyroptosis; Signal Transduction

Islet inflammation and cytokine production are implicated in pancreatic β-cell dysfunction and diabetes pathogenesis. However, we lack therapeutics to protect the insulin-producing β-cells from inflammatory damage. Closing this clinical gap requires the establishment of new disease models of islet inflammation to facilitate screening efforts aimed at identifying new protective agents. Here, we have developed a genetic model of Interleukin-1β (Il-1β)-driven islet inflammation in zebrafish, a vertebrate that allows for non-invasive imaging of β-cells and

Topics: Animals; Animals, Genetically Modified; Apoptosis; Biological Products; Calcium; Coumarins; Cytokines; Disease Models, Animal; Down-Regulation; Glucose; Humans; Hyperglycemia; Inflammation; Insulin-Secreting Cells; Interleukin-1beta; Larva; Macrophages; Mice; Models, Genetic; Time-Lapse Imaging; Transcription, Genetic; Zebrafish

The acute kidney injury(AKI) caused by nephrotoxic drugs contributes to inflammation and oxidative injury in podocytes. Wedelolactone (WED), a natural compound, is found with activities as anti-inflammation, anti-oxidative, anti-free radical,and etc. In this present study, MPC-5 cells were exposed to the nephrotoxic drugs doxorubicin (DOX). The results showed that WED significantly increased the SOD activity, CAT and GSH-Px levels, while significantly decreased the MDA content and ROS levels in DOX-induced MPC-5 cells. WED could also significantly decrease the levels of cytokines IL-6, MCP-1, TNF-α, and TGF-β1. Additionally, the activation and phosphorylation of IκKα, IκBα and NF-κB p65 was inhibited by WED. The co-treatment of PDTC (NF-κB inhibitor) and WED significantly reduced NF-κB p65 phosphorylation. These findings suggested that WED alleviated inflammation and oxidative stress of doxorubicin-induced MPC-5 cells through IκK/IκB/NF-κB signaling pathway.

Topics: Animals; Cell Line; Coumarins; Doxorubicin; I-kappa B Proteins; Inflammation; Mice; NF-kappa B; NF-KappaB Inhibitor alpha; Oxidative Stress; Phosphorylation; Podocytes; Rats; Signal Transduction; Transcription Factor RelA; Tumor Necrosis Factor-alpha

UVB (Ultra-violet B) radiation is one of the major etiological factors in various dermal pathology viz. dermatitis, actinic folliculitis, solar urticaria, psoriasis and cancer among many others. UVB causes toxic manifestation in tissues by inciting inflammatory and tumor promoting events. We have designed this study to assess the anti-inflammatory and anti-tumor promotion effect of Wedelolactone (WDL) a specific IKK inhibitor. Results indicate significant restoration of anti-oxidative enzymes due to WDL treatments. We also found that WDL was effective in mitigating inflammatory markers consisting of MPO (myeloperoxidase), Mast cells trafficking, Langerhans cells suppression and COX 2 expression up regulation due to UVB exposure. We also deduce that WDL presented a promising intervention in attenuating early tumor promotion events caused by UVB exposure as indicated by the results of ODC (Ornithine Decarboxylase), Thymidine assay, Vimentin and VEGF (Vascular-endothelial growth factor) expression. This study was able to provide substantial cues for the therapeutic ability of Wedelolactone against inflammatory and tumor promoting events in murine skin depicting plausible role of NFkB pathway.

Topics: Animals; Carcinogenesis; Cell Communication; Cell Transformation, Neoplastic; Coumarins; DNA; Gene Expression Regulation; Inflammation; Male; Mice; Neovascularization, Pathologic; Oxidative Stress; Polyamines; Skin; Skin Neoplasms; Transcription Factor RelA; Ultraviolet Rays

Cigarette smoke is the leading cause of the development of various lung diseases including lung cancer through triggering oxidant stress and inflammatory responses which contributed to the lesions of normal human bronchial epithelial (NHBE) cell. Wedelolactone (WEL), a natural compound from Eclipta prostrata L., has been found to possess the inhibitive effects on the proliferation and growth of cancers. In the present study, we investigated the effects of WEL on NHBE cell injury induced by cigarette smoke extract (CSE) in vitro. It showed that the pretreatment WEL (2.5-20μM) resulted in a significant protective effect on 10% CSE-induced cell death in NHBE cells. The pretreatment with WEL dose-dependently and significantly reversed the activities of SOD, CAT, GSH and the level of MDA to normal level. We also found that the protein expression levels of COX-2 and ICAM-1 which are related to inflammatory response were remarkably reduced by WEL compared with 10% CSE treatment. Additionally, WEL also reduced the expressions of antioxidases including NAD(P)H dehydrogenase:Quinone 1 (NQO1) and heme oxygenase-1 (HO-1). Moreover, Nrf2 inhibitor all-trans-retinoic acid (ATRA) decreased remarkably their expressions. These results suggest that WEL protects NHBE cell against CSE-induced injury through modulating Nrf2 pathway. Our study indicates that WEL may be a new potential protective agent against CSE-induced lung injury.

Topics: Antioxidants; Bronchi; Cell Line; Complex Mixtures; Coumarins; Cyclooxygenase 2; Dose-Response Relationship, Drug; Epithelial Cells; Gene Expression Regulation; Humans; Inflammation; Intercellular Adhesion Molecule-1; Molecular Structure; NF-E2-Related Factor 2; Oxidants; Respiratory Mucosa; Smoke; Stress, Physiological; Tobacco Products