MH_dev_182

Arsenic decreases antinociceptive activity of paracetamol : possible involvement of serotonergic and endocannabinoid receptors . We assessed whether repeated arsenic exposure can decrease paracetamol-mediated antinociception by modulating serotonergic and endocannabinoid pathways . Rats were preexposed to elemental arsenic ( 4ppm ) as sodium arsenite through drinking water for 28 days . Next day paracetamol ' s ( 400mg / kg , oral ) antinociceptive activity was assessed through formalin-induced nociception . Serotonin content and gene expression of P08908 REA , 5 - Q13049 REA and P21554 REA receptors were evaluated in brainstem and frontal cortex . Arsenic decreased paracetamol-mediated analgesia . DB00316 , but not arsenic , increased serotonin content in these regions . Arsenic attenuated paracetamol-mediated increase in serotonin level . DB00316 did not alter P08908 REA expression , but caused down-regulation of 5 - Q13049 REA and up-regulation of P21554 REA receptors . Arsenic down-regulated these receptors . However , paracetamol-mediated down-regulation of 5 - Q13049 REA was more pronounced . Arsenic did not modify paracetamol ' s effect on P08908 REA expression , but reduced paracetamol-mediated down-regulation of 5 - Q13049 REA and reversed up-regulation of P21554 REA receptors . Results suggest arsenic reduced paracetamol-induced analgesia possibly by interfering with pronociceptive 5 - Q13049 REA and antinociceptive P21554 REA receptors .

P06401 REA activation of extranuclear signaling pathways in regulating p53 expression in vascular endothelial cells . We previously showed that progesterone ( P4 ) inhibited the proliferation of human umbilical vein endothelial cells ( HUVECs ) through a p53 - dependent pathway . Now we investigated further the molecular mechanism underlying the hormone activity . In cultured HUVECs , P4 increased the protein levels of phosphorylated Src ( p-Src ) , P04049 REA , and P29323 REA . The levels of p-Src and p-Src-progesterone receptor complex in HUVECs were increased by P4 treatment . These effects were blocked by pretreatment with a progesterone receptor antagonist , DB00834 SUB . The P4 - induced increase in p53 transactivity was abolished by pretreatment with Src kinase inhibitors . Moreover , administration with cSrc antisense oligonucleotide prevented the P4 - induced increases of the levels of p53 mRNA and protein . These data suggest that P4 - induced up-regulation of p53 might be mediated through activation of cSrc . Pretreatment with Src kinase inhibitors also prevented P4 - induced membrane translocation of Kras and increases of the protein levels of phosphorylated Raf and phosphorylated P29323 REA . Transfection with dominant-negative P28482 REA prevented the P4 - induced increases of protein level and promoter activity of p53 and a decrease of thymidine incorporation . P4 also increased nuclear factor-κB ( NF-κB ) nuclear translocation and NF-κB binding onto the p53 promoter . These effects were abolished by pretreatment with P29323 REA inhibitors . The P4 - induced up-regulation of the p53 promoter activity was prevented by preadministration with dominant-negative P28482 REA or NF-κB inhibitors . Taken together , our data suggest that the cSrc / Kras / P04049 REA / P28482 REA / NF-κB signaling pathway contributes to the P4 - induced up-regulation of p53 in HUVECs . These findings highlight progesterone receptor activation of extranuclear signaling pathways in regulating p53 and cell cycle progression in HUVECs .

New isoflavonoids as inhibitors of porcine P09917 REA . The inhibitory activity of new isoflavonoids on P09917 REA of porcine leukocytes was investigated . Isoflavans ( I ) proved to be stronger inhibitors than isoflavones ( II ) . The isoflavans containing ortho-hydroxy groups in ring A showed the lowest Ki values ( 0.8- 50 microM ) . In comparison , isoflavans with meta-dihydroxy groups exhibited Ki values higher than 150 microM . The effect of commercial antioxidants was tested also on porcine P09917 REA . Butylated hydroxyanisole ( Ki : 25 microM ) and butylated hydroxytoluene ( Ki : 55 microM ) revealed moderate inhibitory activity , whereas L-ascorbic acid , L-ascorbyl palmitate , dl - DB00163 MEN and n-propyl gallate showed weak inhibitory activities ( Ki : 100-260 microM ) .

Ellagic acid inhibits DB00102 MEN - induced vascular smooth muscle cell proliferation and prevents atheroma formation in streptozotocin-induced diabetic rats . Plant-derived polyphenolic compounds have beneficial health effects . In the present study , we determined the ability of ellagic acid ( EA ) to prevent platelet-derived growth factor-BB ( DB00102 MEN ) - induced proliferation of primary cultures of rat aortic smooth muscle cells ( RASMCs ) . We also determined the ability of EA to prevent atherosclerosis in streptozotocin-induced diabetic rats . Proliferation of cells was measured via Alamar Blue assay and through propidium iodide-based cell cycle analysis in flow cytometer . Reactive oxygen species ( ROS ) were measured via 2 ' , 7 ' - dichlorofluorescin diacetate and Amplex red methods . Expression of proliferation markers and activation of kinases were assessed by immunoblot analysis . Cotreatment of primary cultures of RASMCs with 25 μmol / L of EA significantly reduced DB00102 MEN ( 20 ng / ml ) - induced proliferation by blocking S-phase entry . EA effectively blocked PDGF receptor-β ( P09619 REA - β ) tyrosine phosphorylation , generation of intracellular ROS and downstream activation of extracellular signal-regulated kinase 1/2 . It also blocked DB00102 MEN - induced expression of cyclin D1 . Computational molecular docking of EA with the P09619 REA - β - DB00102 MEN complex revealed two putative inhibitor binding sites which showed similar binding energies with the known P09619 REA - β inhibitor AG1295 . In diabetic rats , supplementation of diet with 2 % EA significantly blocked diabetes-induced medial thickness , and lipid and collagen deposition in the arch of aorta . These were assessed through haematoxylin and eosin , Oil Red O and Masson ' s trichome staining , respectively . EA treatment also blocked cyclin D1 expression in medial smooth muscle cells in experimental animals . Thus , EA is effective in reducing atherosclerotic process by blocking proliferation of vascular smooth muscle cells .

Stereospecific interaction of a novel spirosuccinimide type aldose reductase inhibitor , DB05327 MEN , with aldose reductase . P15121 REA ( AR ) is an NADPH-dependent enzyme implicated in diabetic complications . DB05327 MEN [ ( R ) - ( - ) - 2 - ( 4 - bromo - 2 - fluorobenzyl ) -1,2 , 3,4- tetrahydropyrrolo [ 1,2- a ] pyrazine - 4 - spiro - 3 ' - pyrrolidine -1,2 ' , 3,5 ' - tetrone ] is a structurally novel and potent Q9Y4X5 REA with an inhibitor constant ( K ( i ) = 10 ( - ) ( 10 ) M ) 2000 - fold lower than that of its optical antipode ( S-isomer ) . To elucidate the inhibition modes and the stereochemical differences in their inhibitory potencies , we examined the interaction of these R - and S-isomers with AR under physiological conditions . Enzyme kinetic analysis , which was performed by using physiological substrates at 37 degrees C , showed that both isomers selectively act on the E-NADP ( + ) complex in both the forward and reverse reactions of AR . However , fluorometric titration analysis demonstrated that the affinities of the isomers for the E-NADP ( + ) complex are about the same as those for the E-NADPH complex and the apoenzyme . These results suggested that the selective binding to the E-NADP ( + ) complex arises from the predominance of this enzyme form during steady-state turnover rather than from binding specificity . Both the competition with a known active site-directed Q9Y4X5 REA and the protective effect on AR inactivation by N-bromosuccinimide showed that the isomers bind to the active site of the enzyme , but the thermodynamic parameters for the binding to AR indicated that additional hydrogen bonds and / or van der Waals interactions contribute to the energetic stabilization in the E-R-isomer complex . Molecular modeling , together with the deductions from spectroscopic studies , suggested that the succinimide ring and the 4 - bromo - 2 - fluorobenzyl group of the R-isomer are optimally located for formation of a hydrogen-bonding network with AR , and that the latter benzyl group is also effective for the differentiation between AR and aldehyde reductase ( a closely related enzyme ) .

Mechanical stretch increases P08253 REA production in vascular smooth muscle cells via activation of P09619 REA - β / Akt signaling pathway . Increased blood pressure , leading to mechanical stress on vascular smooth muscle cells ( VSMC ) , is a known risk factor for vascular remodeling via increased activity of matrix metalloproteinase ( MMP ) within the vascular wall . This study aimed to identify cell surface mechanoreceptors and intracellular signaling pathways that influence VSMC to produce MMP in response to mechanical stretch ( MS ) . When VSMC was stimulated with MS ( 0-10 % strain , 60 cycles / min ) , both production and gelatinolytic activity of P08253 REA , but not P14780 REA , were increased in a force-dependent manner . MS-enhanced P08253 REA expression and activity were inhibited by molecular inhibition of Akt using Akt siRNA as well as by PI3K / Akt inhibitors , LY293002 and AI , but not by MAPK inhibitors such as PD98059 , SP600125 and SB203580 . MS also increased Akt phosphorylation in VSMC , which was attenuated by AG1295 , a PDGF receptor ( P09619 REA ) inhibitor , but not by inhibitors for other receptor tyrosine kinase including P01133 REA , IGF , and FGF receptors . Although MS activated P09619 REA - α as well as P09619 REA - β in VSMC , MS-induced Akt phosphorylation was inhibited by molecular deletion of P09619 REA - β using siRNA , but not by inhibition of P09619 REA - α . Collectively , our data indicate that MS induces P08253 REA production in VSMC via activation of Akt pathway , that is mediated by activation of P09619 REA - β signaling pathways .

P04150 REA signaling in a bronchial epithelial cell line . Glucocorticoids are an effective anti-inflammatory therapy for the treatment of asthma . The anti-inflammatory effects of glucocorticoids may be due to the inhibition of transcription factors that regulate cytokine synthesis . Because of the potential role of the bronchial epithelium in asthmatic inflammation and the possibility that this cell may be the main target of inhaled glucocorticoids , we have characterized glucocorticoid receptors ( GR ) and GR signaling in the human bronchial epithelial cell line BEAS - 2B . Western blot analysis and radioligand binding studies demonstrated that BEAS - 2B cells have functional GR that bind to dexamethasone ( DB00514 ) ( dissociation constant = 5.6 nM and maximal density of binding sites = 228 + / - 3.3 fmol / mg protein ) . GR were activated by DB00514 as assessed using a glucocorticoid-responsive reporter plasmid . Transfection of BEAS - 2B cells with an activator protein - 1 ( AP - 1 ) reporter construct followed by 12 - O-tetradecanoylphorbol - 13 - acetate ( TPA ) treatment resulted in a fivefold induction of reporter gene activity . Transfection with a nuclear factor ( NF ) - kappa B reporter construct followed by tumor necrosis factor-alpha ( P01375 REA ) treatment resulted in a 10 - fold induction of reporter gene activity . DB00514 ( 10 ( - 7 ) M ) markedly repressed both the induced AP - 1 and NF-kappa B activity . The GR antagonist DB00834 SUB inhibited the repressive effect of DB00514 on P01375 REA - induced NF-kappa B activity by 81 % but only counteracted the repressive effect of DB00514 on TPA-induced AP - 1 activity by 43 % . These studies demonstrate that cross-signaling between AP - 1 and NF-kappa B with GR may explain the anti-inflammatory properties of glucocorticoids in airway epithelial cells .

An endothelial nitric oxide synthase inhibitor aggravates DB03429 - induced acute pancreatitis in rats . To clarify the role of nitric oxide ( NO ) in the development and progression of acute pancreatitis , we investigated the effect of different NO synthase inhibitors and NO donors on experimental pancreatitis in rats . Closed duodenal loop ( DB03429 ) - induced pancreatitis was produced in male Wistar rats , and the animals were treated with normal saline , the NO-synthase substrate L-arginine , the NO donor S-nitroso-N-acetylpenicillamine , aminoguanidine , which is a more powerful inhibitor of inducible NO synthase ( P35228 REA ) than is endothelial NO synthase ( P29474 REA ) , and N-nitro-L-arginine methyl ester ( L-NAME ) , a more powerful inhibitor of P29474 REA than of P35228 REA . All drugs were infused intravenously during a period of 6 or 12 h in each group . Pancreatic tissue was removed at 6 and 12 h after creating the DB03429 . L - DB00125 , S-nitroso-N-acetyl-penicillamine , and aminoguanidine treatment had no effect on the elevation of serum pancreatic enzymes , whereas L-NAME administration significantly exacerbated their elevation . Pathologically , L-NAME treatment resulted in a significantly worse histologic score at 6 and 12 h , especially in terms of the degree of hemorrhage , acinar cell necrosis , and microvascular thrombosis . Addition of L-arginine clearly reversed the effect of L-NAME . Neither the NO substrate nor NO donor could inhibit the progression of hemorrhagic pancreatitis in DB03429 - induced pancreatitis . DB02533 MEN had no effect on the severity of the pancreatitis . We therefore concluded that NO production by P29474 REA may play a significant role in preventing the development and progression of acute pancreatitis .

Dexamethasone inhibits interleukin - 1β - induced matrix metalloproteinase - 9 expression in cochlear cells . OBJECTIVES : To investigate the effect of interleukin ( IL ) - 1β on matrix metalloproteinase ( MMP ) - 9 expression in cochlea and regulation of IL - 1β - mediated P14780 REA expression by dexamethasone and the molecular and signaling mechanisms involved . METHODS : House ear institute-organ of Corti 1 ( HEI-OC 1 ) cells were used and exposed to IL - 1β with / without dexamethasone . P04150 REA antagonist , DB00834 SUB , was used to see the role of dexamethasone . PD98059 ( an extracellular signal-regulated kinases [ ERKs ] inhibitor ) , SB203580 ( a p38 mitogen-activated protein kinases [ MAPK ] inhibitor ) , SP600125 ( a c-Jun N-terminal kinase [ JNK ] inhibitor ) were also used to see the role of MAPKs signaling pathway ( s ) in IL - 1β - induced P14780 REA expression in HEI-OC 1 cells . Reverse transcription-polymerase chain reaction and gelatin zymography were used to measure mRNA expression level of P14780 REA and activity of P14780 REA , respectively . RESULTS : Treatment with IL - 1β - induced the expression of P14780 REA in a dose - and time-dependent manner . IL - 1β ( 1 ng / mL ) - induced P14780 REA expression was inhibited by dexamethasone . Interestingly , p38 MAPK inhibitor , SB203580 , significantly inhibited IL - 1β - induced P14780 REA mRNA and P14780 REA activity . However , inhibition of JNKs and ERKs had no effect on the IL - 1β - induced P14780 REA expression . CONCLUSION : These results suggest that the pro-inflammatory cytokine IL - 1β strongly induces P14780 REA expression via activation of p38 MAPK signaling pathway in HEI-OC 1 cells and the induction was inhibited by dexamethasone .

Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics . Restricting the cytotoxicity of anticancer agents by targeting receptors exclusively expressed on tumor cells is critical when treating infiltrative brain tumors such as glioblastoma multiforme ( GBM ) . GBMs express an P35225 REA receptor ( IL13Rα2 ) that differs from the physiological P24394 REA / IL13R receptor . We developed a regulatable adenoviral vector ( Ad.mhIL-4.TRE.mhIL - 13 - PE ) encoding a mutated human P35225 REA fused to Pseudomonas exotoxin ( mhIL - 13 - PE ) that specifically binds to IL13Rα2 to provide sustained expression , effective anti-GBM cytotoxicity , and minimal neurotoxicity . The therapeutic Ad also encodes mutated human P05112 REA that binds to the physiological P24394 REA / IL13R without interacting with IL13Rα2 , thus inhibiting potential binding of mhIL - 13 - PE to normal brain cells . Using intracranial GBM xenografts and syngeneic mouse models , we tested the Ad.mhIL-4.TRE.mhIL - 13 - PE and two protein formulations , hIL - 13 - PE used in clinical trials ( DB05305 MEN ) and a second-generation mhIL - 13 - PE . DB05305 MEN doubled median survival without eliciting long-term survival and caused severe neurotoxicity ; mhIL - 13 - PE led to ∼ 40 % long-term survival , eliciting severe neurological toxicity at the high dose tested . In contrast , Ad-mediated delivery of mhIL - 13 - PE led to tumor regression and long-term survival in over 70 % of the animals , without causing apparent neurotoxicity . Although DB05305 MEN was originally developed to target GBM , when tested in a phase III trial it failed to achieve clinical endpoints and revealed neurotoxicity . Limitations of DB05305 MEN include its short half-life , which demanded frequent or continued administration , and binding to P24394 REA / IL13R , present in normal brain cells . These shortcomings were overcome by our therapeutic Ad , thus representing a significant advance in the development of targeted therapeutics for GBM .

Thromboxane A ( 2 ) generation , in the absence of platelet P23219 REA activity , in patients with and without atherothrombotic myocardial infarction . BACKGROUND : DB00945 MENMAX DB00945 MEN ' s therapeutic action is via inhibition of platelet cyclooxygenase 1 ( P23219 REA ) thromboxane A2 ( TxA 2 ) production . The aim of this study was to evaluate TxA 2 production , in the absence of platelet P23219 REA activity , in coronary atherosclerotic heart disease patients with and without atherothrombotic myocardial infarction ( MI ) . METHODS AND RESULTS : TxA 2 production , in the absence of platelet P23219 REA activity , was evaluated in 44 patients taking aspirin on 3 commercially available assays that detect metabolites of TxA 2 in the urine . Two assays measure urine 11 - dehydro-thromboxane B2 ( TxB 2 ) alone and 1 measures urine 11 - dehydro-TxB 2 plus 11 - dehydro -2,3- dinor-TxB 2 . Platelet P23219 REA inhibition was confirmed on < 10 % platelet aggregation in response to ≥ 1 mmol / L arachidonic acid . Median urine 11 - dehydro-TxB 2 was no different in those with and without a diagnosis of atherothrombotic MI ( 325 vs . 311 pg / mg creatinine , P= 0.59 via polyclonal ELISA ) and ( 312 vs . 244 pg / mg creatinine , P= 0.11 via LC-MS / MS ) . Median urine 11 - dehydro-TxB 2 plus 11 - dehydro -2,3- dinor-TxB 2 , however , was higher in those with vs . those without a diagnosis of atherothrombotic MI ( 1,035 vs . 606 pg / mg creatinine , P= 0.03 via monoclonal ELISA ) . CONCLUSIONS : Differences in TxA 2 production , in the absence of platelet P23219 REA activity , between those with vs . without atherothrombotic MI were not observed when TxA 2 generation was assessed on 11 - dehydro-TxB 2 production alone ( polyclonal ELISA or LC-MS / MS ) , but differences were observed when TxA 2 generation was assessed using 11 - dehydro-TxB 2 plus 11 - dehydro -2,3- dinor-TxB 2 ( monoclonal ELISA ) . These findings highlight important differences between different commercially available assays for TxA 2 generation and suggest that 11 - dehydro -2,3- dinor-TxB 2 may be critical to the biology of atherothrombosis .

P15121 REA regulates TGF-beta 1 - induced production of fibronectin and type IV collagen in cultured rat mesangial cells . AIM : To study the effects of aldose reductase ( AR ) on production of fibronectin and type IV collagen in rat mesangial cells ( MsC ) . METHODS : The vector , pcDNA 3 - AR , was constructed based on pET - 15b - AR . Lipofect AMINE was used for stable transfection and G418 was used for selecting positive clones . DB02712 and zopolrestat were added for suppressing the activity of AR , respectively . The production of fibronectin and type IV collagen and the activation of Smads and MAPK signal transduction pathway were analysed by western blot and AP - 1 activity was analysed by electrophoretic mobility shift assays ( EMSA ) . RESULTS : The normal MsC showed increased expression of fibronectin and type IV collagen with stimulation of TGF-beta 1 . Compared with the normal MsC , the MsC pre-incubated with Q9Y4X5 REA showed reduced expression ( P < 0.05 ) and the AR-transfected MsC showed increased expression ( P < 0.05 ) . The normal MsC showed activation of P29323 REA , JNK and p38 with stimulation of TGF-beta 1 , while the activation of JNK and p38 was inhibited in the MsC pre-incubated with Q9Y4X5 REA and only the activation of JNK was enhanced in the AR-transfected MsC . The normal MsC showed enhanced AP - 1 activity with the stimulation of TGF-beta 1 , and similarly the activity was inhibited in the MsC pre-incubated with Q9Y4X5 REA and was more enhanced in the AR transfected MsC . CONCLUSION : AR can regulate the expression of fibronectin and type IV collagen with the stimulation of TGF-beta 1 in MsC , which may have relations with the activation of JNK-MAPK and p38 - MAPK signalling pathways and AP - 1 .

Characterization of DB05005 MEN - B , an orally bioavailable antagonist of the P51686 REA chemokine receptor , for treatment of inflammatory bowel disease . The chemokine system represents a diverse group of G protein-coupled receptors responsible for orchestrating cell recruitment under both homeostatic and inflammatory conditions . Chemokine receptor 9 ( P51686 REA ) is a chemokine receptor known to be central for migration of immune cells into the intestine . Its only ligand , O15444 REA , is expressed at the mucosal surface of the intestine and is known to be elevated in intestinal inflammation . To date , there are no reports of small-molecule antagonists targeting P51686 REA . We report , for the first time , the discovery of a small molecule , DB05005 MEN - B , which is an orally bioavailable , selective , and potent antagonist of human P51686 REA . DB05005 MEN - B inhibited P51686 REA - mediated Ca ( 2 + ) mobilization and chemotaxis on Molt - 4 cells with IC ( 50 ) values of 5.4 and 3.4 nM , respectively . In the presence of 100 % human serum , DB05005 MEN - B inhibited P51686 REA - mediated chemotaxis with an IC ( 50 ) of 33 nM , and the addition of α1 - acid glycoprotein did not affect its potency . DB05005 MEN - B inhibited chemotaxis of primary P51686 REA - expressing cells to O15444 REA with an IC ( 50 ) of 6.8 nM . DB05005 MEN - B was an equipotent inhibitor of O15444 REA - directed chemotaxis of both splice forms of P51686 REA ( CCR 9A and CCR 9B ) with IC ( 50 ) values of 2.8 and 2.6 nM , respectively . DB05005 MEN - B also inhibited mouse and rat P51686 REA - mediated chemotaxis . Inhibition of P51686 REA with DB05005 MEN - B results in normalization of Crohn ' s disease such as histopathology associated with the P01375 REA ( ΔARE ) mice . Analysis of the plasma level of drug associated with this improvement provides an understanding of the pharmacokinetic / pharmacodynamic relationship for P51686 REA antagonists in the treatment of intestinal inflammation .

P04150 REA antagonism disrupts the reconsolidation of social reward-related memories in rats . Reconsolidation is the process whereby consolidated memories are destabilized upon retrieval and restabilized to persist for later use . Although the neurobiology of the reconsolidation of both appetitive and aversive memories has been intensively investigated , reconsolidation of memories of physiologically relevant social rewards has received little attention . Social play , the most characteristic social behaviour displayed by young mammals , is highly rewarding , illustrated by the fact that it can induce conditioned place preference ( CPP ) . Here , we investigated the role of signalling mechanisms implicated in memory processes , including reconsolidation , namely glucocorticoid , mineralocorticoid , DB01221 glutamatergic and P21554 REA cannabinoid receptors , in the reconsolidation of social play-induced CPP in rats . Systemic treatment with the glucocorticoid receptor antagonist mifepristone before , but not immediately after , retrieval disrupted the reconsolidation of social play-induced CPP . DB00834 SUB did not affect social play-induced CPP in the absence of memory retrieval . Treatment with the DB01221 receptor antagonist MK - 801 modestly affected the reconsolidation of social play-induced CPP . However , the reconsolidation of social play-induced CPP was not affected by treatment with the mineralocorticoid and P21554 REA cannabinoid receptor antagonists spironolactone and rimonabant , respectively . We conclude that glucocorticoid neurotransmission mediates the reconsolidation of social reward-related memories in rats . These data indicate that the neural mechanisms of the reconsolidation of social reward-related memories only partially overlap with those underlying the reconsolidation of other reward-related memories .

Cannabinoid receptors and T helper cells . We have reported that injection of marijuana cannabinoids , such as Delta ( 9 ) - tetrahydrocannabinol ( THC ) , into mice , followed by infection with Legionella pneumophila ( Lp ) , suppresses the development of cell-mediated immunity T helper 1 ( Th1 ) activity . These effects are accompanied by suppression of interleukin ( IL ) - 12 and interferon ( IFN ) gamma production and enhancement of P05112 REA production suggesting THC-induced T helper cell biasing . In the current report , other T helper cell biasing mechanisms were studied . Mice were injected with THC followed 18 h later by a challenge infection with Lp . Two-hour post-infection , spleens were removed and analyzed for mRNA to either IL - 12Rbeta2 or P23771 REA gene products . The results showed that THC suppressed IL - 12Rbeta2 but increased P23771 REA . Receptor antagonists for P21554 REA ( SR141716A , SR1 ) and CB2 ( SR144528 , SR2 ) were also injected to analyze the involvement of cannabinoid receptors . It was determined that SR1 attenuated the THC suppression of IL - 12Rbeta2 , while SR2 attenuated the increase in P23771 REA mRNA . These results suggest that THC suppresses Th1 biasing activity such as IL - 12Rbeta2 by a P21554 REA mediated mechanism and enhances the Th2 biasing activity , P23771 REA , by a CB2 mechanism . This dichotomy of receptor involvement might result from differential expression and / or signaling function of P21554 REA and CB2 on Th1 and Th2 cells .

Dual ligands targeting dopamine D2 and serotonin P08908 REA receptors as new antipsychotical or anti-Parkinsonian agents . Psychiatric disorders like schizophrenia and neurodegenerative diseases like Parkinson ' s disease are associated with poly-factorial pathogenic mechanisms , with several neurotransmitter systems closely involved . In addition to the cerebral dopaminergic ( DA ) system , the serotoninergic ( 5 - HT ) system also plays a crucial role in regulating psychoemotional , cognitive and motor functions in the central nervous system ( CNS ) . Among the large 5 - HT receptor family , accumulating data have revealed new insights into the therapeutic benefit of the P08908 REA receptor in treating various CNS disorders , especially schizophrenia and Parkinson ' s disease . The present review discusses the advance of dual agents with mixed actions at the dopamine D2 and serotonin P08908 REA receptors in the treatment of these diseases . Aripiprazole was the only marketed drug with dual D2 and P08908 REA profile . It is a partial D2 and P08908 REA receptor agonist and has been prescribed as an atypical antipsychotical drug . Two other drugs DB06016 MEN and Pardoprunox are being investigated in clinic . Most of the other candidate compounds , including DB04888 , Sarizotan , Mazapertine succinate , PF - 217830 , and Adoprazine were discontinued due to either non-optimal pharmacokinetic properties or insufficient therapeutical efficacy . Although much effort has been done to highlight the advantages of the P08908 REA and D2 dual approach , it has to be pointed out that many of these drugs showed poly-pharmacological profile by targeting many other receptors and / or transporters besides the D2 and P08908 REA receptors . In this regard , ' pure ' compounds exclusively acting on the D2 and P08908 REA receptors are highly needed to further validate this approach . Meanwhile , safety concerns and in vivo pharmacokinetic alerts should also be implanted to the drug design art early .

Inhibition of cyclooxygenase - 2 elicits a P21554 REA - mediated decrease of excitatory transmission in rat P00915 REA hippocampus . Cannabinoid receptor ( P21554 REA ) ligands decrease excitatory and inhibitory transmission in the hippocampus , but the influence of endogenously formed cannabinoids ( eCBs ) on basal excitatory transmission remains uncertain . Here , we investigated the influence of eCBs on synaptic transmission in P00915 REA hippocampus using the slice preparation . Blockade of P21554 REA with the selective receptor antagonists SR141716 ( rimonabant ) or AM251 augmented synaptic responses evoked upon stimulation of the Schaffer collaterals . This effect persisted in the presence of bicuculline or CGP 55845 to block GABA ( A ) or GABA ( B ) receptors , revealing a tonic eCB influence on excitatory transmission . Selective inhibition of cyclooxygenase - 2 ( P35354 REA ) with meloxicam or NS - 398 decreased excitatory responses partly in a P21554 REA - dependent manner , independently of GABA ( A ) transmission . Paired-pulse paradigms suggested a presynaptic P21554 REA mechanism to decrease glutamate release . Inhibition of P23219 REA or other routes of eCB degradation did not affect synaptic transmission . We conclude that P35354 REA regulates the formation of P21554 REA ligands that decrease hippocampal excitatory transmission .

Endocannabinoids as autoregulatory signaling molecules : coupling to nitric oxide and a possible association with the relaxation response . Endocannabinoid signaling processes are present in diverse organisms and in organisms 500 million years divergent in evolution . Cannabinoid receptor - 1 expression ( P21554 REA ) , anandamide , and anandamide amidase have been found in invertebrates . Furthermore , this signaling system is coupled to constitutive nitric oxide synthase ( P29474 REA ) - derived nitric oxide ( NO ) release in both vertebrates and invertebrates , thereby regulating neural , immune , and vascular-like functions in these divergent organisms . In human endothelial cells from various blood vessels , P21554 REA immunoreactive components are present as is its coupling to anandamide-stimulated P29474 REA - derived NO production , which exerts an autoregulatory role on P29474 REA release . The modulation of vascular diameter and vascular tone represents a crucial point of interest in these pathways , and interactions between NO and the sympathetic nerve system are of importance , i . e , norepinephrine . Here , a possible association of NO and endocannabinoid signaling with the relaxation response , a physiological counterpart of the stress response , may exist .

P60568 REA suppression by 2 - arachidonyl glycerol is mediated through peroxisome proliferator-activated receptor gamma independently of cannabinoid receptors 1 and 2 . 2 - Arachidonyl glycerol ( 2 - AG ) is an endogenous arachidonic acid derivative that binds cannabinoid receptors P21554 REA and CB2 and is hence termed an endocannabinoid . 2 - AG also modulates a variety of immunological responses , including expression of the autocrine / paracrine T cell growth factor interleukin ( IL ) - 2 . The objective of the present studies was to determine the mechanism responsible for P60568 REA suppression by 2 - AG . Because of the labile properties of 2 - AG , 2 - AG ether , a nonhydrolyzable analog of 2 - AG , was also used . Both 2 - AG and 2 - AG ether suppressed P60568 REA expression independently of P21554 REA and CB2 , as demonstrated in leukocytes derived from P21554 REA / CB2 - null mice . Moreover , we demonstrated that both 2 - AG and 2 - AG ether treatment activated peroxisome proliferator-activated receptor gamma ( PPARgamma ) , as evidenced by forced differentiation of 3T3 - Q9NUQ9 cells into adipocytes , induction of aP2 mRNA levels , and activation of a PPARgamma-specific luciferase reporter in transiently transfected 3T3 - Q9NUQ9 cells . Consequently , the putative role of PPARgamma in P60568 REA suppression by 2 - AG and 2 - AG ether was examined in Jurkat T cells . Concordant with PPARgamma involvement , the PPARgamma-specific antagonist 2 - chloro - 5 - nitro-N - ( 4 - pyridyl ) - benzamide ( T0070907 ) blocked 2 - AG - and 2 - AG ether-mediated P60568 REA suppression . Likewise , 2 - AG suppressed the transcriptional activity of two transcription factors crucial for P60568 REA expression , nuclear factor of activated T cells and nuclear factor kappaB , in the absence but not in the presence of T0070907 . 2 - AG treatment also induced PPARgamma binding to a Q07869 REA response element in activated Jurkat T cells . Together , the aforementioned studies identify PPARgamma as a novel intracellular target of 2 - AG , which mediates the suppression of P60568 REA by 2 - AG in a manner that is independent of P21554 REA and / or CB2 .

Reduction of human monocytic cell neurotoxicity and cytokine secretion by ligands of the cannabinoid-type CB2 receptor . 1 Two cannabinoid receptors , P21554 REA and CB2 , have been identified . The P21554 REA receptor is preferentially expressed in brain , and the CB2 receptor in cells of leukocyte lineage . We identified the mRNA for the P21554 REA receptor in human neuroblastoma SH-SY 5Y cells , and the mRNA and protein for the CB2 receptor in human microglia and THP - 1 cells . 2 Delta ( 9 ) - and Delta (8 ) - tetrahydrocannabinol ( THC ) were toxic when added directly to SH-SY 5Y neuroblastoma cells . The toxicity of Delta ( 9 ) - THC was inhibited by the P21554 REA receptor antagonist SR141716A but not by the CB2 receptor antagonist SR144528 . The endogenous ligand anandamide was also toxic , and this toxicity was enhanced by inhibitors of its enzymatic hydrolysis . 3 The selective CB2 receptor ligands JWH - 015 and indomethacin morpholinylamide ( BML - 190 ) , when added to THP - 1 cells before stimulation with lipopolysaccharide ( LPS ) and P01579 REA , reduced the toxicity of their culture supernatants to SH-SY 5Y cells . JWH - 015 was more effective against neurotoxicity of human microglia than THP - 1 cells . The antineurotoxic activity of JWH - 015 was blocked by the selective CB2 receptor antagonist SR144528 , but not by the P21554 REA receptor antagonist SR141716A . This activity of JWH - 015 was synergistic with that of the P09917 REA ( 5 - P28300 REA ) inhibitor REV 5901 . 4 Cannabinoids inhibited secretion of IL - 1beta and tumor necrosis factor-alpha ( P01375 REA ) by stimulated THP - 1 cells , but these effects could not be directly correlated with their antineurotoxic activity . 5 Specific CB2 receptor ligands could be useful anti-inflammatory agents , while avoiding the neurotoxic and psychoactive effects of P21554 REA receptor ligands such as Delta ( 9 ) - THC .

Q07869 REA agonists for the treatment of cardiovascular disease in patients with diabetes . Diabetes is a complex disease defined by hyperglycaemia ; however , strong associations with abdominal obesity , hypertension and dyslipidaemia contribute to the high risk of cardiovascular disease . Although aggressive glycaemic control reduces microvascular complications , the evidence for macrovascular complications is less certain . The theoretical benefits of the mode of action of peroxisome proliferator-activated receptor ( Q07869 REA ) agonists are clear . In clinical practice , Q07869 REA - α agonists such as fibrates improve dyslipidaemia , while Q07869 REA - γ agonists such as thiazolidinediones improve insulin resistance and diabetes control . However , although these agents are traditionally classed according to their target , they have different and sometimes conflicting clinical benefit and adverse event profiles . It is speculated that this is because of differing properties and specificities for the Q07869 REA receptors ( each of which targets specific genes ) . This is most obvious in the impact on cardiovascular outcomes - - in clinical trials pioglitazone appeared to reduce cardiovascular events , whereas rosiglitazone potentially increased the risk of myocardial infarction . The development of a dual Q07869 REA - α / γ agonist may prove beneficial in effectively managing glycaemic control and improving dyslipidaemia in patients with type 2 diabetes . Yet , development of agents such as muraglitazar and tesaglitazar has been hindered by various serious adverse events . DB08915 MEN , a balanced dual Q07869 REA - α / γ agonist , is currently the most advanced in clinical development and has shown promising results in phase II clinical trials with beneficial effects on glucose and lipid variables . A phase III study , ALECARDIO , is ongoing and will establish whether improvements in laboratory test profiles translate into an improvement in cardiovascular outcomes .