phenanthrenes and Edema

Interruption of the Warburg effect - the observation that un-stimulated macrophages reprogram their core metabolism from oxidative phosphorylation toward aerobic glycolysis to become pro-inflammatory M1 macrophages upon stimulation - is an emerging strategy for the treatment of cancer and anti-inflammatory diseases such as rheumatoid arthritis. We studied this process with view to the discovery of novel therapeutics, and found that tylophorine-based compounds targeted a ribonucleoprotein complex containing caprin-1 and mRNAs of c-Myc and HIF-1α in LPS/IFN-γ stimulated Raw264.7 cells, diminished the protein levels of c-Myc and HIF-1α, and consequently downregulated their targeted genes that are associated with the Warburg effect, as well as the pro-inflammatory iNOS and COX2. The tylophorine-based compound DBQ 33b significantly meliorated the severity and incidence of type II collagen-monoclonal antibody-induced rheumatoid arthritis and diminished gene expressions of c-Myc, HIF-1α, iNOS, COX2, TNFα, and IL-17A in vivo. Moreover, pharmacological inhibition of either c-Myc or HIF-1α exhibited similar effects as the tylophorine-based compound DBQ 33b, even though inhibition of c-Myc reversed the induction of iNOS and COX2 in LPS/IFN-γ stimulated Raw264.7 cells to a lesser degree. Therefore, simultaneous inhibition of both c-Myc and HIF-1α is efficacious for anti-inflammation in vitro and in vivo and merits further study.

Topics: Alkaloids; Animals; Anti-Inflammatory Agents; Arthritis, Rheumatoid; Cell Cycle Proteins; Cyclooxygenase 2; Edema; Female; Hypoxia-Inducible Factor 1, alpha Subunit; Indolizines; Interferon-gamma; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Nitric Oxide Synthase Type II; Phenanthrenes; Proto-Oncogene Proteins c-myc; Rats, Sprague-Dawley; RAW 264.7 Cells; RNA, Messenger; Tumor Necrosis Factor-alpha

The objective of this present study was to develop and evaluate the triptolide-loaded nanostructured lipid carriers (TPL-NLCs) for transdermal drug delivery system (TDDS). TPL-NLCs was prepared with emulsification technique and optimized by central composite design of a response surface methodology (CCD-RSM). The optimized TPL-NLCs were spherical and physically stable with the average size of 139.6.0 ± 2.53 nm and Zeta potential of -36.03 ± 2.41 mV. The encapsulation efficiency and drug loading were 97.15% ± 9.46 and 10.35% ± 1.12, respectively. Moreover, the in vitro release study showed that TPL-NLCs had a sustained release profiles and the in vitro penetration study indicated that TPL-NLCs could effectively penetrate into skin. Besides, the methodology of skin-blood synchronous microdialysis was established to evaluate the pharmacokinetics of TPL-NLCs in vivo and the results displayed that the TPL concentration in skin was higher than that in blood. And TPL-NLCs presented a remarkable effect of decreasing knee edema, inhibiting inflammation by regulating the levels of TNF-α, IL-1β and IL-6, which indicated that TPL-NLCs was a promising topical administration in treatment of edema and inflammation associated with rheumatoid arthritis (RA).

Topics: Administration, Cutaneous; Animals; Arthritis, Rheumatoid; Delayed-Action Preparations; Disease Models, Animal; Diterpenes; Drug Carriers; Drug Delivery Systems; Drug Liberation; Edema; Epoxy Compounds; Immunosuppressive Agents; Lipids; Male; Microdialysis; Nanostructures; Particle Size; Phenanthrenes; Rats; Rats, Sprague-Dawley; Skin Absorption

Dihydrotanshinone (DHT), one of the major ingredients of Salvia miltiorrhiza Bunge (Danshen), displays many bioactivities. However, the activity and underlying mechanism of DHT in anti-inflammation have not yet been elucidated. In this study, we investigated the anti-inflammatory activity and molecular mechanism of action of DHT both in vitro and in vivo. Our data showed that DHT significantly decreased the release of inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β in lipopolysaccharide (LPS)-stimulated RAW264.7 cells, THP-1 cells, and bone marrow-derived macrophages (BMDMs), and altered the expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). In addition, flow cytometry results indicated that DHT reduced the calcium influx, and generation of reactive oxygen species (ROS), and nitric oxide (NO) generation in LPS-stimulated RAW264.7 cells. Moreover, DHT suppressed the transcription of nuclear factor-κB (NF-κB), the expressions of NF-κB proteins, and nuclear translocation of NF-κB/p65, thereby suggesting that the NF-κB pathway played a role in the anti-inflammatory action of DHT. In addition, DHT attenuated LPS-challenged activator protein-1 (AP-1) activity, resulting from interference of the mitogen-activated protein kinase (MAPK) pathway. The molecular docking simulation of DHT to toll-like receptor 4 (TLR4) suggested that DHT binds to the active sites of TLR4 to block TLR4 dimerization, which was further corroborated by cellular thermal shift assay and co-immunoprecipitation (Co-IP) experiments. Furthermore, the recruitment of myeloid differentiation primary response gene 88 (MyD88) and the expression of transforming growth factor-b (TGF-b)-activated kinase 1 (p-TAK1) were disturbed by the inhibition of TLR4 dimerization. Thus, investigating the molecular mechanism of DHT indicated that TLR4-MyD88-NF-κB/MAPK signaling cascades were involved in the anti-inflammatory activity of DHT in vitro. In in vivo mouse models, DHT significantly ameliorated LPS-challenged acute kidney injury, inhibited dimethylbenzene-induced mouse ear oedema, and rescued LPS-induced sepsis in mice. Taken together, our results indicated that DHT exhibited significant anti-inflammatory activity both in vitro and in vivo, suggesting that DHT may be a potential therapeutic agent for inflammatory diseases.

Topics: Acute Kidney Injury; Animals; Anti-Inflammatory Agents; Cytokines; Dimerization; Edema; Furans; HEK293 Cells; Humans; Lipopolysaccharides; Male; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; NF-kappa B; Phenanthrenes; Quinones; RAW 264.7 Cells; Sepsis; THP-1 Cells; Toll-Like Receptor 4; Xylenes

Podocytes are important to glomerular filtration barrier integrity and maintenance of size selectivity in protein filtration in the kidney. Although there is evidence to suggest that triptolide has direct protective effects on podocyte injuries, the mechanism mediating this process remains largely unexplored. In this study, we found triptolide suppresses podocyte p53 and GADD45B expression in vivo and in vitro. We used our previously developed in vivo zebrafish model of inducible podocyte-targeted injury and found that triptolide or the inhibition of p53 and gadd45ba with morpholino (MO) alleviated metronidazole (MTZ) induced edema in zebrafish, while the overexpression of gadd45ba in podocytes blocked the protective effect of triptolide and p53 MO on podocyte injury in zebrafish. Further study showed that p53 directly transactivated GADD45B. Triptolide inhibited p53 binding to the GADD45B promoter and subsequent GADD45B transcription. We further demonstrated that p53 may indirectly regulate GADD45B expression via NF-κB signaling. Taken together, our findings demonstrated that triptolide maintained glomerular barrier function via the inhibition of p53-NF-κB-GADD45B signaling, which provides a new understanding of the antiproteinuric effects of triptolide in glomerular diseases.

Topics: Animals; Antigens, Differentiation; Diterpenes; Edema; Epoxy Compounds; Glomerular Filtration Barrier; Metronidazole; NF-kappa B; Phenanthrenes; Podocytes; Protein Binding; Puromycin Aminonucleoside; Signal Transduction; Tumor Suppressor Protein p53; Yolk Sac; Zebrafish; Zebrafish Proteins

Benefits from thrombolysis with recombinant tissue plasminogen activator (rt-PA) after ischemic stroke remain limited due to a narrow therapeutic window, low reperfusion rates, and increased risk of hemorrhagic transformations (HT). Experimental data showed that rt-PA enhances the post-ischemic activation of poly(ADP-ribose)polymerase (PARP) which in turn contributes to blood-brain barrier injury. The aim of the present study was to evaluate whether PJ34, a potent PARP inhibitor, improves poor reperfusion induced by delayed rt-PA administration, exerts vasculoprotective effects, and finally increases the therapeutic window of rt-PA. Stroke was induced by thrombin injection (0.75 UI in 1 μl) in the left middle cerebral artery (MCA) of male Swiss mice. Administration of rt-PA (0.9 mg kg

Topics: Animals; Edema; Endothelial Cells; Hemorrhage; Infarction, Middle Cerebral Artery; Inflammation; Male; Mice; Neuroprotective Agents; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Proteolysis; Recombinant Proteins; Reperfusion; Stroke; Thrombosis; Tissue Plasminogen Activator; Treatment Outcome; Vasospasm, Intracranial

The present study was designed to explore the influence of water extracts of Atractylodes lancea rhizomes on the toxicity and anti-inflammatory effects of triptolide (TP). A water extract was prepared from A. lancea rhizomes and co-administered with TP in C57BL/6 mice. The toxicity was assayed by determining serum biochemical parameters and visceral indexes and by liver histopathological analysis. The hepatic CYP3A expression levels were detected using Western blotting and RT-PCR methods. The data showed that the water extract of A. lancea rhizomes reduced triptolide-induced toxicity, probably by inducing the hepatic expression of CYP3A. The anti-inflammatory effects of TP were evaluated in mice using a xylene-induced ear edema test. By comparing ear edema inhibition rates, we found that the water extract could also increase the anti-inflammatory effects of TP. In conclusion, our results suggested that the water extract of A. lancea rhizomes, used in combination with TP, has a potential in reducing TP-induced toxicity and enhancing its anti-inflammatory effects.

Topics: Animals; Anti-Inflammatory Agents; Atractylodes; Cytochrome P-450 CYP3A; Cytochrome P-450 Enzyme System; Diterpenes; Edema; Enzyme Induction; Epoxy Compounds; Gene Expression Regulation; Herb-Drug Interactions; Liver; Male; Mice; Mice, Inbred C57BL; Phenanthrenes; Plant Extracts; Plants, Medicinal; Rhizome; Water

To date, Kaposi sarcoma has not been mentioned among the adverse effects of triptolide/tripdiolide, ethyl acetate extracts or polyglycosides of the Chinese herbal remedy Tripterygium wilfordii Hook F.. A patient was diagnosed with rheumatoid arthritis at the age of 29 years. She underwent treatment with corticosteroids, methotrexate and gold sodium thiosulfate, and was chronically taking ketoprofen. At the age of 59 years she started to take a powder (≈2 g/day) from a Chinese physician for treatment of rheumatoid arthritis. This powder was supplied to her regularly for 10 years. At the age of 69 years, multiple soft, violaceous to dark-red patches, plaques, nodules and blisters of varying sizes appeared on a background of severely edematous skin on her legs, and later on her arms. Biopsy specimens of the leg lesions were diagnostic for human herpesvirus 8-associated Kaposi sarcoma. Triptolide (235 µg/1 g) and tripdiolide were found in the Chinese powder by the use of Liquid Chromatography Electrospray Ionization Mass Spectrometry. Administration of the powder was stopped and medication with paclitaxel was introduced. General condition of the patient improved and skin lesions diminished significantly.. This case indicates a possible association between triptolide/tripdiolide chronic intake and development of human herpesvirus 8-associated Kaposi sarcoma. Triptolide/tripdiolide could contribute to development of Kaposi sarcoma by reactivation of latent human herpesvirus 8, permitted by immunosuppression induced by triptolide.

Topics: Adult; Aged; Arthritis, Rheumatoid; Diterpenes; Edema; Epoxy Compounds; Female; Humans; Immunosuppressive Agents; Leg; Middle Aged; Phenanthrenes; Powders; Sarcoma, Kaposi; Skin Neoplasms

Current study was conducted to identify constituents of Taxus wallichiana Zucc. that might be responsible for its folk use in anti-inflammatory conditions. Taxusabietane A was isolated from the bark extract of Taxus wallichiana Zucc. Taxusabietane A was analyzed for in-vitro and in-vivo anti-inflammatory activities using Lipoxygenase (LOX) inhibition assay and carrageenan-induced paw edema model. Taxusabietane A revealed considerable LOX inhibitory activity with the IC(50) value being 57 ± 0.31. Standard compound Baicalein showed the IC(50) value being 22.1 ± 0.03 μM. Taxusabietane A also showed significant (5 and 10 mg/kg) anti-inflammatory activity induced by carrageenan. However, this study highlighted the potential of Taxusabietane A to be further explored as a new lead compound for management of conditions associated with inflammation.

Topics: Animals; Anti-Inflammatory Agents; Carrageenan; Disease Models, Animal; Edema; Female; Flavanones; Lipoxygenase Inhibitors; Male; Phenanthrenes; Phytotherapy; Plant Bark; Plant Extracts; Rats; Rats, Wistar; Taxus

Disruption of blood-brain barrier (BBB) and edema formation play a key role in the development of neurological dysfunction after cerebral ischemia. In this study, the effects of Tanshinone IIA (Tan IIA), one of the active ingredients of Salvia miltiorrhiza root, on the BBB and brain edema after transient middle cerebral artery occlusion in rats were examined. Our study demonstrated that Tan IIA reduced brain infarct area, water content in the ischemic hemisphere. Furthermore, Tan IIA significantly decreased BBB permeability to Evans blue, suppressed the expression of intercellular adhesion molecule-1 (ICAM-1), matrix metalloproteinase-9 (MMP-9), inhibited the degradation of tight junction proteins zonula occludens-1 (ZO-1) and Occludin. These results demonstrated that Tan IIA was effective for attenuating the extent of brain edema formation in response to ischemia injury in rats, partly by Tan IIA's protective effect on the BBB. Our results may have implications in the treatment of brain edema in cerebral ischemia.

Topics: Abietanes; Animals; Blood-Brain Barrier; Brain; Edema; Evans Blue; Infarction, Middle Cerebral Artery; Intercellular Adhesion Molecule-1; Ischemic Attack, Transient; Matrix Metalloproteinase 9; Membrane Proteins; Occludin; Permeability; Phenanthrenes; Phytotherapy; Plant Extracts; Plant Roots; Rats; Reperfusion Injury; Salvia miltiorrhiza; Tight Junctions; Water

9,10-Dihydro-2,5-dimethoxyphenanthrene-1,7-diol (RSCL-0520) is a phenanthrene isolated from Eulophia ochreata, one of the Orchidaceae family, known by local tradition to exhibit medicinal properties. However, no anti-inflammatory activity or any molecular mechanisms involved have been reported or elucidated. Here, for the first time, we evaluate the anti-inflammatory properties of RSCL-0520 on responses induced by lipopolysaccharide (LPS) and mediated via Toll-like receptors (TLRs).. The in vitro anti-inflammatory activities of RSCL-0520 were investigated in LPS-stimulated monocytic cells, measuring activation of cytokine and inflammatory genes regulated by nuclear factor-kappaB (NF-kappaB). Tumour necrosis factor (TNF)-alpha levels in serum following LPS stimulation in mice and carrageenan-induced paw oedema in rats were used as in vivo models.. Pretreatment with RSCL-0520 effectively inhibited LPS-induced, TLR4-mediated, NF-kappaB-activated inflammatory genes in vitro, and reduced both LPS-induced TNF-alpha release and carrageenan-induced paw oedema in rats. Treatment with RSCL-0520 reduced LPS-stimulated mRNA expression of TNF-alpha, COX-2, intercellular adhesion molecule-1, interleukin (IL)-8 and IL-1beta, all regulated through NF-kappaB activation. RSCL-0520, however, did not interfere with any cellular processes in the absence of LPS.. RSCL-0520 blocked signals generated by TLR4 activation, as shown by down-regulation of NF-kappaB-regulated inflammatory cytokines. The inhibitory effect involved both MyD88-dependent and -independent signalling cascades. Our data elucidated the molecular mechanisms involved, and support the search for plant-derived TLR antagonists, as potential anti inflammatory agents.

Topics: Animals; Cell Line; Dose-Response Relationship, Drug; Down-Regulation; Edema; Female; Humans; Inflammation; Inflammation Mediators; Lipopolysaccharides; Male; Mice; Mice, Inbred BALB C; Monocytes; Orchidaceae; Phenanthrenes; Rats; Rats, Wistar; Signal Transduction; Toll-Like Receptors; Tumor Necrosis Factor-alpha

This study examined the effects of tanshinone derivatives (tanshinone I, cryptotanshinone, 15,16-dihydrotanshinone I) on prostaglandin (PG) and nitric oxide (NO) metabolism in an attempt to establish their anti-inflammatory mechanisms and to present a scientific rationale for the use of Salvia miltiorrhiza (danshen) in inflammatory conditions. From lipopolysaccharide-treated RAW 264.7 cells, cyclooxygenase-2 (COX-2)-mediated PGE2 production was inhibited by tanshinone I, cryptotanshinone and 15,16-dihydrotanshinone I, while only cryptotanshinone and 15,16-dihydrotanshinone I inhibited inducible NO synthase (iNOS)-mediated NO synthesis at 1-50 microM. Particularly, cryptotanshinone was found to be a down-regulator of proinflammatory molecule expression, including COX-2 and iNOS. The electrophoretic mobility shift assay showed that cryptotanshinone and 15,16-dihydrotanshinone I also inhibited the activation of the transcription factors, such as nuclear transcription factor-kappaB and activator protein-1. Moreover, cryptotanshinone exhibited in vivo anti-inflammatory activity against carrageenan-induced paw edema in rats. Overall, these results provide additional scientific rationale for the anti-inflammatory use of danshen in Chinese medicine. Especially, cryptotanshinone and 15,16-dihydrotanshinone I are important constituents.

Topics: Abietanes; Animals; Anti-Inflammatory Agents, Non-Steroidal; Carrageenan; Cell Line; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Dinoprostone; Disease Models, Animal; Dose-Response Relationship, Drug; Down-Regulation; Edema; Electrophoretic Mobility Shift Assay; Furans; Lipopolysaccharides; Macrophages; Male; Mice; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Phenanthrenes; Plant Roots; Quinones; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza

Cyclooxygenase-2 (COX-2) is a key enzyme that catalyzes the biosynthesis of prostaglandins from arachidonic acid and plays a critical role in some pathologies including inflammation, neurodegenerative diseases and cancer. Cryptotanshinone is a major constituent of tanshinones, which are extracted from the medicinal herb Salvia miltiorrhiza Bunge, and has well-documented antioxidative and anti-inflammatory effects. This study confirmed the remarkable anti-inflammatory effect of cryptotanshinone in the carrageenan-induced rat paw edema model. Since the action of cryptotanshinone on COX-2 has not been previously described, in the present study, we further examined the effect of cryptotanshinone on cyclooxygenase activity in the exogenous arachidonic acid-stimulated insect sf-9 cells, which highly express human COX-2 or human COX-1, and on cyclooxygenases expression in human U937 promonocytes stimulated by lipopolysaccharide (LPS) plus phorbolmyristate acetate (PMA). Cryptotanshinone reduced prostaglandin E2 synthesis and reactive oxygen species generation catalyzed by COX-2, without influencing COX-1 activity in cloned sf-9 cells. In PMA plus LPS-stimulated U937 cells, cryptotanshinone had negligible effects on the expression of COX-1 and COX-2, at either a mRNA or protein level. These results demonstrate that the anti-inflammatory effect of cryptotanshinone is directed against enzymatic activity of COX-2, not against the transcription or translation of the enzyme.

Topics: Animals; Anti-Inflammatory Agents; Arachidonic Acid; Blotting, Western; Carrageenan; Cell Line; Cyclooxygenase 1; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Dinoprostone; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Edema; Gene Expression Regulation, Enzymologic; Hindlimb; Humans; Molecular Structure; Phenanthrenes; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Reverse Transcriptase Polymerase Chain Reaction; Salvia; Spodoptera; U937 Cells

Triptolide (TP) has been shown to have anti-inflammatory, immunosuppressive, anti-fertility and anti-neoplastic activity. However, its clinical use was restricted to some extent due to its serious toxicity. The possible mechanism for triptolide-induced hepatotoxicity was related to reactive oxygen species (ROS) inducing lipid peroxidation and DNA damage. The development of controlled release delivery strategies could lead to significant advantages in the clinical use of these drugs to decreasing the toxicity. Thus, the present study was focused on the preparation and some characterization of triptolide-loaded solid lipid nanoparticle (SLN) and the measurements of anti-inflammatory activities and the hepatotoxicity of TP-SLN. The carrageenan-induced rat paw edema experiment indicated that the anti-inflammatory activities of TP-SLN were stronger than those of free triptolide. Orally administration of TP-SLN 0.2 or 0.4 mg/kg per day did not cause mortality within the period of observation. In contrast, free triptolide at different doses had caused partial death. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were significantly elevated in the free triptolide-treated group whereas they did not significantly change in TP-SLN-treated mice. The free triptolide increased malondialdehyde (MDA) level and decreased activities of superoxide dismutase (SOD) and total glutathione peroxidase (GSH-Px) in the liver homogenates. However, these phenomena were not found in TP-SLN-treated mice. The results of histopathological evaluation revealed a protective effect of SLN on vacuolation, edema, inflammatory infiltration and necrosis caused by triptolide. Furthermore, TP-SLN did not change Bcl/Bax protein ratio or decrease FasL expression in liver cells. These results suggest that SLN delivery system can enhance the anti-inflammatory activity of triptolide meanwhile has a protective effect against triptolide-induced hepatotoxicity. The toxicity of TP-SLN to other tissues is under investigation.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Delayed-Action Preparations; Diterpenes; Dose-Response Relationship, Drug; Edema; Epoxy Compounds; Liver; Male; Mice; Nanostructures; Phenanthrenes; Rats; Rats, Wistar

Triptolide (TP), which has potent immunosuppressive effects, anti-inflammatory and severe toxicity on digestive, urogenital, blood circulatory system, was used as a model drug in this study. The aim of this study was to investigate the anti-inflammatory effect of complete Freund's adjuvant-induced arthritis in rats treated with TP-loaded poly(D,L-lactic acid) (PLA) nanoparticles (TP-PLA-NPs) by gavage. TP-PLA-NPs were prepared by the modified spontaneous emulsification solvent diffusion method (modified-SESD). Nanoparticles were shown to be small particle size (149.7 nm), low polydispersity index (0.088), a fine spherical shape with smooth surfaces determined by dynamic light scattering (DLS) and transmission electron microscope (TEM). Encapsulation efficiency and the in vitro release of TP from nanoparticles were measured by the reverse phase high-performance liquid chromatography (RP-HPLC). The in vitro release profile of TP from nanoparticles exhibited a typical biphasic release phenomenon, namely initial burst release and consequently slow release. The potential therapeutic arthritic method of TP-PLA-NPs was established. The results obtained in experiments indicated that TP-PLA-NPs significantly inhibited the adjuvant-induced arthritis, and had preferable anti-inflammatory effect with the long-time administration.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Chemistry, Pharmaceutical; Diterpenes; Drug Combinations; Edema; Epoxy Compounds; Lactic Acid; Male; Nanostructures; Phenanthrenes; Polyesters; Polymers; Rats; Rats, Wistar

Arachidonic acid (AA) mainly released from the cell membrane by phospholipase A(2) (PLA(2)) is converted to eicosanoids by the action of cyclooxygenase (COX) and lipoxygenase (LO). In order to find the specific inhibitors of AA metabolism especially PLA(2) and COX-2, 300 plant extracts were evaluated for their inhibitory activity on PGD(2) production from cytokine-induced mouse bone marrow-derived mast cells in vitro. From this screening procedure, the methanol extract of Salvia miltiorrhiza was found to inhibit PGD(2) production and the ethyl acetate subfraction gave the strongest inhibition of five subfractions tested. From this ethyl acetate subfraction, an activity-guided isolation finally gave tanshinone I as an active principle. This investigation deals with the effects of tanshinone I on AA metabolism from lipopolysaccharide (LPS)-induced RAW 264.7 cells and in vivo antiinflammatory activity. Tanshinone I inhibited PGE(2) formation from LPS-induced RAW macrophages (IC(50) = 38 microM). However, this compound did not affect COX-2 activity or COX-2 expression. Tanshinone I was found to be an inhibitor of type IIA human recombinant sPLA(2)(IC(50) = 11 microM) and rabbit recombinant cPLA(2) (IC(50) = 82 microM). In addition, tanshinone I showed in vivo antiinflammatory activity in rat carrageenan-induced paw oedema and adjuvant-induced arthritis.

Topics: Abietanes; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arachidonic Acid; Arthritis, Experimental; Blotting, Western; Carrageenan; Cyclooxygenase 2; Dose-Response Relationship, Drug; Edema; Humans; Inhibitory Concentration 50; Isoenzymes; Male; Mast Cells; Membrane Proteins; Mice; Phenanthrenes; Phytotherapy; Plant Extracts; Prostaglandin D2; Prostaglandin-Endoperoxide Synthases; Rabbits; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza

The anti-inflammatory activity of abietic acid, a diterpene isolated from Pimenta racemosa var. grissea (Myrtaceae), was evaluated in-vivo and in-vitro. This compound significantly inhibited rat paw oedema induced by carrageenan in a time- and dose-dependent manner, and mouse ear oedema induced by 12-O-tetradecanoylphorbol acetate, after oral or topical administration. The inhibition of myeloperoxidase enzyme showed that its topical activity was influenced by neutrophil infiltration into the inflamed tissues (ears). In addition, the effect of abietic acid on some macrophage functions was analysed in-vitro. Non-toxic concentrations of abietic acid inhibited prostaglandin E2 (PGE2) production in lipopolysaccharide-treated macrophages, whereas nitrite, tumour necrosis factor alpha and interleukin-1beta production were only weakly affected by this diterpene. PGE2 production from A23187-stimulated macrophages was only inhibited at high doses (100 microM) and it failed to modify leukotriene C4 production. These results indicate that abietic acid exerts in-vivo anti-inflammatory activity after oral or topical administration and has partial ability to prevent the production of some inflammatory mediators.

Topics: Abietanes; Administration, Oral; Administration, Topical; Animals; Cytokines; Diterpenes; Edema; Fibrinolytic Agents; Inflammation; Macrophages; Male; Mice; Neutrophil Infiltration; Phenanthrenes; Plant Extracts; Rats; Rats, Wistar

The anti-inflammatory action of nonapeptide fragments of uteroglobin or lipocortin I known as antiflammins, was tested in the carrageenan or phospholipase A2 rat paw oedema model. The development of carrageenan-induced oedema in rats was significantly inhibited during the early and late phases of the oedema by the local administration of antiflammins 1 and 2. However, the peptides were not able to inhibit phospholipase A2-induced oedema. The time course of the anti-oedematous activity of nonapeptides after intradermal carrageenan injection may be attributed to their effect on mast cell degranulation and accumulation and activation of leukocytes. Naja naja phospholipase A2 exhibited strong histamine release-inducing activity, which may have contributed to the rat paw oedema induction. Surprisingly, antiflammins had a limited but significant inhibitory effect on histamine secretion.

Topics: Animals; Annexin A1; Aristolochic Acids; Carrageenan; Cell Degranulation; Chlorpheniramine; Dexamethasone; Disease Models, Animal; Edema; Elapid Venoms; Elapidae; Histamine Release; Indomethacin; Inflammation; Injections, Intradermal; Male; Mast Cells; Peptide Fragments; Phenanthrenes; Phospholipases A; Phospholipases A2; Rats; Rats, Sprague-Dawley; Uteroglobin

Triptolide (Tri) was isolated from Tripterygium wilfordii Hook f. Tri 0.1-0.2 mg.kg-1 sc or 0.15-0.3 mg.kg-1 ig inhibited markedly the increased vascular permeability induced by ip 0.7% HAc in mice. Tri 0.05-0.1 mg.kg-1 ip or 0.15-0.3 mg.kg-1 ig inhibited hind paws swelling induced by sc 0.15 ml carrageenan and also inhibited the same swelling induced by sc 2.5% formaldehyde 0.1 ml in rats. Tri 0.05-0.1 mg.kg-1 ip inhibited markedly proliferation of granuloma induced by sc implantation of cotton-pellets in rats, but 0.2 mg.kg-1 ip can not inhibit the same swelling induced by sc 0.15 ml carrageenan in adrenalectomized rats. Tri 0.2 mg.kg-1 ip decreased markedly weight of thymus. Tri 0.2 mg.kg-1 ip, but 0.1 mg.kg-1 ip did not reduced the content of ascorbic acid of adrenal gland in rats. Tri 0.2 mg.kg-1 ip did not decrease the pro-staglandin E content in inflammatory tissues. These results indicate that high dose of Tri can stimulate the pituitary-adrenal axis.

Topics: Adrenal Glands; Adrenalectomy; Animals; Anti-Inflammatory Agents, Non-Steroidal; Ascorbic Acid; Capillary Permeability; Carrageenan; Diterpenes; Edema; Epoxy Compounds; Granuloma, Foreign-Body; Male; Mice; Phenanthrenes; Prostaglandins E; Rats

Injection of phospholipase A2 (PLA2) in the rat skin produced a significant rise in oedema, which was inhibited by the simultaneous coinjection of aristolochic acid (100 micrograms), mepacrine (100 micrograms) and p-bromophenacyl bromide (10 micrograms). Indomethacin, nordihydroguaiaretic acid and WEB 2086 were without inhibitory effect on this model, whereas dexamethasone (5 mg/kg, p.o.) and coinjection of chlorpheniramine (20 micrograms) inhibited the oedema formation by more than 60%. Dose-dependent histamine release by rat peritoneal cells was induced by PLA2 and by lysophosphatidylserine and this effect could be antagonized by aristolochic acid and mepacrine. Apomorphine, previously reported to be an antagonist at lysophospholipid receptors, was able to inhibit the histamine release by mast cells as well as the oedema formation in rat skin. Taken all together, these results suggest that lysophospholipids, produced by the action of PLA2, are the mediators for the histamine release in rat peritoneal cells and could play an important role in the early oedema development in rat skin after PLA2 administration.

Topics: Animals; Aristolochic Acids; Cytoplasmic Granules; Dexamethasone; Edema; Histamine Release; In Vitro Techniques; Indomethacin; Lysophospholipids; Male; Mast Cells; Phenanthrenes; Phospholipases A; Phospholipases A2; Quinacrine; Rats; Rats, Inbred Strains; Skin

The phenanthrylethanols 2a-d were obtained by reduction of the acetyl derivatives 1a-d and converted, through the phenanthrylethyl halides 3a-d and 4b, into the nitriles 5a-d, whose acid hydrolysis afforded the acids of the title, 6a-d. The antiinflammatory activity of these acids was measured on the carrageenin-induced edema and found as 1/3 (6a), 1/43 (6b), 1/5 (6c), and 1/7 (6d) of that of fenbufen.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Carrageenan; Chemical Phenomena; Chemistry; Edema; Male; Phenanthrenes; Propionates; Rats; Rats, Inbred Strains

A neutral-active, Ca2+-dependent phospholipase A2 (PLA2) purified 11,000-fold from human synovial fluid (HSF) induced edema when injected into the mouse foot pad. The edema produced by HSF-PLA2 was dose-dependent and was positively correlated with the dose-dependent in vitro expression of PLA2 activity. Maximum edema was achieved within 45 min after the injection and persisted for at least 6 h. Aristolochic acid [8-methoxy-6-nitrophenanthro(3,4-d)-1,3-dioxole-5-carboxylic acid], a major chemical component derived from various species of Aristolochia plant, produced a dose-dependent inhibition of in vitro phospholipid hydrolysis by HSF-PLA2, porcine pancreatic PLA2, snake venom (Naja naja) PLA2, and PLA2 isolated from human platelet. The sensitivity of these PLA2s to inhibition by aristolochic acid varied markedly: HSF-PLA2 greater than N. naja PLA2 greater than human platelet PLA2 greater than porcine pancreatic PLA2. The inhibition of HSF-PLA2 by aristolochic acid was independent of substrate concentration (18-144 microM) and Ca2+ concentration (0.1-4.0 mM). These observations indicate that inhibition of HSF-PLA2 by aristolochic acid may result from direct interaction with the enzyme. When aristolochic acid was mixed with HSF-PLA2 and then injected into the mouse foot pad, edema was inhibited in a dose-dependent manner and was positively correlated with in vitro inhibition of PLA2 activity. Alkylation of HSF-PLA2 with p-bromophenacyl bromide concomitantly inhibited both enzyme and edema-inducing activity. These results clearly demonstrate that the neutral-active, Ca2+-dependent PLA2 isolated from human synovial fluid is proinflammatory and that catalytic activity is positively correlated with in vivo proinflammatory effects.

Topics: Acetophenones; Animals; Aristolochic Acids; Edema; Enzyme Activation; Female; Humans; Male; Mice; Phenanthrenes; Phospholipases; Synovial Fluid

Aristolochic acid, an alkaloid from the plant Aristolochia species, interacts with the major basic phospholipase A2 from Vipera russelli venom. It is an uncompetitive inhibitor with a Ki of 9.9 X 10(-4)M when phosphatidylcholine is used as substrate. The inhibition of direct and indirect hemolysis is higher compared to the inhibition of phosphatidylcholine hydrolysis. Edema-inducing activity of Vipera russelli phospholipase A2 is inhibited by aristolochic acid when injected either as a mixture or separately. Both i.m. and i.p. administration of aristolochic acid following phospholipase injection are equally effective in inhibiting edema. The alkaloid inhibits the edema-inducing activity as soon as it reaches the site, but does not aid in recovery. Aristolochic acid failed to inhibit other pathological activities of the enzyme.

Topics: Animals; Aristolochic Acids; Edema; Mice; Phenanthrenes; Phosphatidylcholines; Phospholipases; Phospholipases A; Phospholipases A2; Viper Venoms

A basic phospholipase A2 (PLA2) enzyme, TFV PL-X (pI 9.2) and two acidic PLA2 enzymes, TFV PL-Ia (pI 4.9) and TFV PL-Ib (pI 4.5) were purified from Trimeresurus flavoviridis venom on CM-Sephadex C-25 and QAE-Sephadex A-25 columns, respectively. The basic enzyme exists as a monomer, whereas the acidic enzymes are dimers. These enzymes differ in properties such as molecular weight, Km, optimum pH and temperature and pharmacological properties. The basic enzyme hydrolysed purified phospholipids in the order of PC greater than PE greater than PS greater than PI = 0, while for TFV PL-Ia and TFV PL-Ib the order was PC greater than PE greater than PS = PI = 0. TFV PL-X was comparatively more toxic, with an LD50 value of 4.2 micrograms/g (i.p.), while the acidic PLA2 enzymes had LD50 values above 8 micrograms/g (i.p.). All three enzymes induced edema when injected into the mouse foot pad. Aristolochic acid, an alkaloid (8-methoxy-6-nitrophenanthro(3,4-d)-1,3-dioxole-5-carboxylic acid) from the medicinal plant Aristolochia radix, interacts with these PLA2 enzymes. It is a competitive inhibitor with varying affinity when PC is used as substrate. Aristolochic acid inhibits direct and indirect hemolytic activity, as well as edema-inducing activity, of TFV PL-X, but fails to neutralize the lethal potency of the enzyme. On the other hand, it inhibits direct and indirect lytic activity of TFV PL-Ia and TFV PL-Ib only at 10-fold higher concentrations and it enhances the edema-inducing activity of these enzymes. Such effects of aristolochic acid indicates that (1) different mechanisms may be involved in the edema-inducing activity of PLA2 enzymes and (2) catalytic and pharmacological sites are separate on the PLA2 molecule.

Topics: Animals; Aristolochic Acids; Biological Assay; Chemical Phenomena; Chemistry; Chromatography, Gel; Edema; Hemolysis; Molecular Weight; Phenanthrenes; Phosphatidylcholines; Phospholipases; Phospholipases A; Phospholipases A2; Viper Venoms

Topics: Abietanes; Animals; Anti-Infective Agents; Anti-Inflammatory Agents; Capillary Permeability; Cell Movement; Edema; Female; Leukocytes; Male; Mice; Phenanthrenes; Rats

Topics: Animals; Anti-Inflammatory Agents; Arthritis; Carrageenan; Edema; Formaldehyde; Inflammation; Phenanthrenes; Rats; Saponins