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Xu B, Guo Y, Zhang M, Zhang R, Chen D, Zhang Q, Xiao J, Xu K, Li N, Qiu Y, Zhu H, Niu J, Zhang X, Fang Q. Central and peripheral modulation of gastrointestinal transit in mice by DN-9, a multifunctional opioid/NPFF receptor agonist. Neurogastroenterol Motil 2020; 32:e13848. [PMID: 32281198 DOI: 10.1111/nmo.13848] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/22/2019] [Revised: 02/05/2020] [Accepted: 03/19/2020] [Indexed: 02/06/2023]
Abstract
BACKGROUND The nonapeptide DN-9 functions as a multifunctional agonist to opioid and neuropeptide FF (NPFF) receptors and exhibits antinociceptive effects at the central and peripheral levels. METHODS The effects of DN-9 on small and colonic intestinal transit were evaluated using the upper gastrointestinal (GI) transit test and colonic bead expulsion assay, respectively. Opioid and NPFF receptor antagonists were used to investigate the mechanisms of DN-9-induced GI inhibition. Furthermore, the agonism of the DN-9 analog [Phg9 ]-DN-9 to opioid and NPFF receptors was tested by the cAMP assay. KEY RESULTS Intracerebroventricular administration of DN-9 dose-dependently slowed upper GI transit and colonic expulsion via mu- and kappa-opioid receptors in the brain, independent of the delta-opioid receptor. Similarly, intraperitoneal injection of DN-9 dose-dependently inhibited GI propulsion via the peripheral opioid receptors. DN-9-induced GI transit inhibitions were significantly aggravated by the NPFF receptor antagonist RF9. Moreover, the DN-9 analog [Phg9 ]-DN-9, an agonist at mu-, delta-, and kappa-opioid receptors but not NPFF receptors, inhibited GI more potently than DN-9. In addition, intracerebroventricular NPFF significantly attenuated the central inhibitory effects induced by [Phg9 ]-DN-9 and morphine. However, central and peripheral injections of NPFF or RF9 almost had no significant effects on GI transit by itself. CONCLUSION AND INFERENCES Intracerebroventricular and intraperitoneal administrations of DN-9 inhibit GI transit via opioid receptors in mice by central and peripheral mechanisms, respectively. In addition, the NPFF agonism of DN-9 possesses antiopioid effects on GI transit, which might explain the reduced constipation at the antinociceptive doses.
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Affiliation(s)
- Biao Xu
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Yuanyuan Guo
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Mengna Zhang
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Run Zhang
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Dan Chen
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Qinqin Zhang
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Jian Xiao
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Kangtai Xu
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Ning Li
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Yu Qiu
- Department of Pharmacology and Chemical Biology, Institute of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Hanwen Zhu
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Jiandong Niu
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Xiaoyu Zhang
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
| | - Quan Fang
- Key Laboratory of Preclinical Study for New Drugs of Gansu Province, and Institute of Physiology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
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Kumar K, Goyal R, Mudgal A, Mohan A, Pasha S. YFa and analogs: Investigation of opioid receptors in smooth muscle contraction. World J Gastroenterol 2011; 17:4523-31. [PMID: 22110284 PMCID: PMC3218144 DOI: 10.3748/wjg.v17.i40.4523] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/19/2011] [Revised: 06/16/2011] [Accepted: 06/23/2011] [Indexed: 02/06/2023] Open
Abstract
AIM: To study the pharmacological profile and inhibition of smooth muscle contraction by YFa and its analogs in conjunction with their receptor selectivity.
METHODS: The effects of YFa and its analogs (D-Ala2) YFa, Y (D-Ala2) GFMKKKFMRF amide and Des-Phe-YGGFMKKKFMR amide in guinea pig ileum (GPI) and mouse vas deferens (MVD) motility were studied using an isolated tissue organ bath system, and morphine and DynA (1-13) served as controls. Acetylcholine was used for muscle stimulation. The observations were validated by specific antagonist pretreatment experiments using naloxonazine, naltrindole and norbinaltorphimine norBNI.
RESULTS: YFa did not demonstrate significant inhibition of GPI muscle contraction as compared with morphine (15% vs 62%, P = 0.0002), but moderate inhibition of MVD muscle contraction, indicating the role of κ opioid receptors in the contraction. A moderate inhibition of GPI muscles by (Des-Phe) YFa revealed the role of anti-opiate receptors in the smooth muscle contraction. (D-Ala-2) YFa showed significant inhibition of smooth muscle contraction, indicating the involvement of mainly δ receptors in MVD contraction. These results were supported by specific antagonist pretreatment assays.
CONCLUSION: YFa revealed its side-effect-free analgesic properties with regard to arrest of gastrointestinal transit. The study provides evidences for the involvement of κ and anti-opioid receptors in smooth muscle contraction.
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Moulédous L, Mollereau C, Zajac JM. Opioid-modulating properties of the neuropeptide FF system. Biofactors 2010; 36:423-9. [PMID: 20803521 DOI: 10.1002/biof.116] [Citation(s) in RCA: 55] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/11/2010] [Accepted: 06/30/2010] [Indexed: 12/21/2022]
Abstract
Opioid receptors are involved in the control of pain perception in the central nervous system together with endogenous neuropeptides, termed opioid-modulating peptides, participating in a homeostatic system. Neuropeptide FF (NPFF) and related peptides possess anti-opioid properties, the cellular mechanisms of which are still unclear. The purpose of this review is to detail the phenomenon of cross-talk taking place between opioid and NPFF systems at the in vivo pharmacological level and to propose cellular and molecular models of functioning. A better knowledge of the mechanisms underlying opioid-modulating properties of NPFF has potential therapeutic interest for the control of opioid functions, notably for alleviating pain and/or for the treatment of opioid abuse.
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Affiliation(s)
- Lionel Moulédous
- Institut de Pharmacologie et de Biologie Structurale, CNRS UMR5089, Université de Toulouse, 205 route de Narbonne, Toulouse CEDEX 04, France
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Nakajima H, Goto H, Azuma YT, Fujita A, Takeuchi T. Functional interactions between the SK2 channel and the nicotinic acetylcholine receptor in enteric neurons of the guinea pig ileum. J Neurochem 2007; 103:2428-38. [PMID: 17953675 DOI: 10.1111/j.1471-4159.2007.04960.x] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
Abstract
The neurotransmitter acetylcholine (ACh) plays a critical role in gastrointestinal function. The role of the small conductance Ca2+-activated K+ (SK) channel in ACh release was examined using myenteric plexus preparations of guinea pig ileum. Apamin, an inhibitor of the SK channel, significantly enhanced nicotine-induced ACh release, but neither electrical field stimulation- nor 5-hydroxytryptamine-induced ACh release, suggesting that SK channels might be selectively involved in the regulation of nicotine-induced ACh release. Therefore, we investigated the distribution of SK2 and SK3 subunits and the interaction between SK2 channels and nicotinic ACh receptors (nAChRs) in the guinea pig ileum. The immunoreactivity of SK2 subunits was located in enteric neuronal cells. Furthermore, SK2-immunoreactive cells stained with an antibody for choline acetyltransferase, a marker for cholinergic neurons, and with an antibody for the alpha3/5 subunits of nAChR. In contrast, immunoreactivity of SK3 subunits was not found in enteric neurons. A co-immunoprecipitation assay with Triton X-100-soluble membrane fractions prepared from the ileum revealed an association of the SK2 subunit with the alpha3/5 subunits of nAChR. These results suggest that SK2 channels negatively regulate the excitation of enteric neurons via functional interactions with nAChRs.
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Affiliation(s)
- Hidemitsu Nakajima
- Laboratory of Veterinary Pharmacology, Graduate School of Life and Environmental Science, Osaka Prefecture University, Naka-ku, Sakai, Japan
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Roumy M, Lorenzo C, Mazères S, Bouchet S, Zajac JM, Mollereau C. Physical association between neuropeptide FF and micro-opioid receptors as a possible molecular basis for anti-opioid activity. J Biol Chem 2007; 282:8332-42. [PMID: 17224450 DOI: 10.1074/jbc.m606946200] [Citation(s) in RCA: 46] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/31/2022] Open
Abstract
Neuropeptide FF (NPFF) modulates the opioid system by exerting functional anti-opioid activity on neurons, the mechanism of which is unknown. By using a model of SH-SY5Y cells, we recently postulated that anti-opioid activity likely takes place upstream from the signaling cascade, suggesting that NPFF receptors could block opioid receptors by physical interaction. In the present study, fluorescence techniques were used to monitor the physical association and the dynamic of NPFF2 and micro-opioid (MOP) receptors tagged with variants of the green fluorescent protein. Importantly, cyan fluorescent protein-tagged NPFF2 receptors retained their capacity to antagonize opioid receptors. Fluorescence resonance energy transfer (FRET) and coimmunoprecipitation studies indicate that NPFF and MOP receptors are close enough to generate a basal FRET signal. The opioid agonist Tyr-D-Ala-Gly-NMe-Phe-Gly-ol disrupts by 20-30% this FRET signal, mainly because it concomitantly induces 40% internalization of receptors. In contrast, the NPFF analog 1DMe significantly increases by 10-15% the basal FRET signal, suggesting an association between both receptors. In addition, 1DMe reduces, by half, MOP receptor internalization, indicating that, besides a functional blockade of opioid receptors, the NPFF analog also inhibits their internalization. Finally, as a first report showing the modulation of the mobility of a G-protein-coupled receptor by another one, fluorescence recovery after photobleaching analysis reveals that 1DMe modifies the lateral diffusion of MOP receptors in the cell membrane, changing them from a confined to a freely diffusing state. By promoting NPFF-MOP receptor heteromerization, 1DMe could disrupt the domain organization of MOP receptors in the membrane, resulting in a reduction of opioid response.
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Affiliation(s)
- Michel Roumy
- Institut de Pharmacologie et Biologie Structurale, CNRS UMR 5089, 205 route de Narbonne, 31077 Toulouse cedex 04, France
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Kersanté F, Mollereau C, Zajac JM, Roumy M. Anti-opioid activities of NPFF1 receptors in a SH-SY5Y model. Peptides 2006; 27:980-9. [PMID: 16488058 DOI: 10.1016/j.peptides.2005.07.025] [Citation(s) in RCA: 32] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/22/2005] [Accepted: 07/04/2005] [Indexed: 11/28/2022]
Abstract
In order to elucidate the mechanisms of the neuronal anti-opioid activity of Neuropeptide FF, we have transfected the SH-SY5Y neuroblastoma cell line, which expresses mu- and delta-opioid receptors, with the human NPFF1 receptor. The SH1-C7 clone expresses high affinity NPFF1 receptors in the same range order of density as opioid receptors. Similarly to the opioids, acute stimulation with the NPFF1 agonist NPVF inhibits adenylyl cyclase activity and voltage-gated (N-type) Ca2+ currents and enhances the intracellular Ca2+ release triggered by muscarinic receptors activation. In contrast, preincubation of cells with NPVF decreases the response to opioids on both calcium signaling, thus reproducing the cellular anti-opioid activity described in neurons. SH1-C7 cells are therefore a suitable model to investigate the interactions between NPFF and opioid receptors.
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Affiliation(s)
- Flavie Kersanté
- Institut de Pharmacologie et de Biologie Structurale, CNRS, UMR 5089, 205 route de Narbonne, 31077 Toulouse cedex 04, France
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Okishio Y, Takeuchi T, Fujita A, Suenaga K, Fujinami K, Munakata S, Hata F. Examination of the role of cholinergic myenteric neurons with the impairment of neural reflexes in the ileum of c-kit mutant mice. J Smooth Muscle Res 2005; 41:49-60. [PMID: 15855739 DOI: 10.1540/jsmr.41.49] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
Our previous study showed that impairment of ascending and descending neural reflexes in the ileum of the c-kit mutant, W/W(V), mice is due to a loss of interstitial cells of Cajal present at the myenteric plexus region (ICC-MY) in the mutant. In the present study, cholinergic interneurons were thought to be involved in these pathways, since hexamethonium, an antagonist of the nicotinic ACh receptor, significantly inhibited both neural reflexes in wild type mice. Therefore, we examined whether the loss of ICC-MY affects cholinergic interneurons involved in these pathways. Immunohistochemistry with anti-choline acetyltransferase revealed that there was no difference in the numbers of immunopositive cells in the myenteric plexus region between the wild type and mutant mice. In addition, there was no difference in the extent of spontaneous and EFS-evoked ACh release from longitudinal muscle with myenteric plexus preparations between the wild type and mutant mice. Exogenously added nicotine induced contraction or relaxation of ileal circular muscle in the absence or presence of atropine, respectively, to a similar extent in both the wild type and mutant mice. These results suggest that loss of ICC-MY resulted in an impairment of the ascending and descending reflex pathways at the step before activation of cholinergic interneurons.
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Affiliation(s)
- Yutaka Okishio
- Department of Veterinary Pharmacology, Graduate School of Agriculture and Life Sciences, Osaka Prefecture University, Sakai, Japan
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Fang Q, Guo J, Chang M, Chen LX, Chen Q, Wang R. Neuropeptide FF receptors exert contractile activity via inhibition of nitric oxide release in the mouse distal colon. Peptides 2005; 26:791-7. [PMID: 15808909 DOI: 10.1016/j.peptides.2004.12.009] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/09/2004] [Revised: 12/10/2004] [Accepted: 12/14/2004] [Indexed: 11/29/2022]
Abstract
Neuropeptide FF (NPFF) and NPVF, two closely NPFF related peptides, have different affinities for the two NPFF receptors (NPFF1 and NPFF2). To assess the peripheral effects of NPFF receptors in the gastrointestinal tract motility, NPFF and NPVF were tested in the mouse isolated distal colon. Both NPFF (1-15 microM) and NPVF (1-15 microM) dose-dependently caused significant colonic contractions. Pre-treatment with the putative NPFF antagonist, BIBP3226 (30 microM) abolished the contractile responses to the two neuropeptides (3 microM). They had no additional contractile activities in colonic preparations contracted by Nomega-nitro-L-arginine (30 microM). Moreover, the contractions of these two neuropeptides were weakened by L-arginine (2 mM). The responses to NPFF (5 microM) and NPVF (5 microM) were not modified by atropine or naloxone (1 microM). Furthermore, NPFF (1 microM) and NPVF (1 microM) did not influence the contractive responses to acetylcholine (0.1-10 microM), morphine (1 microM) or nociceptin (0.1 microM). These data suggest that NPFF and NPVF cause contractions of the mouse distal colon via their NPFF receptors and this effect is mediated by NO but not by cholinergic pathways, independently from opioid system. In addition, the isolated bioassay may be applied as a simple parameter to characterize the potential NPFF agonists and antagonists.
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Affiliation(s)
- Quan Fang
- (a)Department of Biochemistry and Molecular Biology, School of Life Science, Lanzhou University, 222 Tian Shui South Road, Lanzhou 730000, PR China
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Takeuchi T, Fujinami K, Goto H, Fujita A, Taketo MM, Manabe T, Matsui M, Hata F. Roles of M2 and M4 Muscarinic Receptors in Regulating Acetylcholine Release From Myenteric Neurons of Mouse Ileum. J Neurophysiol 2005; 93:2841-8. [PMID: 15574798 DOI: 10.1152/jn.00986.2004] [Citation(s) in RCA: 33] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
We investigated the subtype of presynaptic muscarinic receptors associated with inhibition of acetylcholine (ACh) release in the mouse small intestine. We measured endogenous ACh released from longitudinal muscle with myenteric plexus (LMMP) preparations obtained from M1–M5 receptor knockout (KO) mice. Electrical field stimulation (EFS) increased ACh release in all LMMP preparations obtained from M1–M5 receptor single KO mice. The amounts of ACh released in all preparations were equal to that in the wild-type mice. Atropine further increased EFS-induced ACh release in the wild-type mice. Unexpectedly, atropine also increased, to a similar extent, EFS-induced ACh release to the wild-type mice in all M1–M5 receptor single KO mice. In M2 and M4 receptor double KO mice, the amount of EFS-induced ACh release was equivalent to an atropine-evoked level in the wild-type mouse, and further addition of atropine had no effect. M2 receptor immunoreactivity was located in both smooth muscle cells and enteric neurons. M4 receptor immunoreactivity was located in the enteric neurons, being in co-localization with M2 receptor immunoreactivity. These results indicate that both M2 and M4 receptors mediate the muscarinic autoinhibition in ACh release in the LMMP preparation of the mouse ileum, and loss of one of these subtypes can be compensated functionally by a receptor that remained. M1, M3, and M5 receptors do not seem to be involved in this mechanism.
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Affiliation(s)
- Tadayoshi Takeuchi
- Department of Veterinary Pharmacology, Graduate School of Agriculture and Life Science, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai Osaka 599-8531, Japan.
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Mollereau C, Mazarguil H, Zajac JM, Roumy M. Neuropeptide FF (NPFF) analogs functionally antagonize opioid activities in NPFF2 receptor-transfected SH-SY5Y neuroblastoma cells. Mol Pharmacol 2005; 67:965-75. [PMID: 15608144 DOI: 10.1124/mol.104.004614] [Citation(s) in RCA: 55] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022] Open
Abstract
To elucidate the mechanism of the cellular antiopioid activity of neuropeptide FF (NPFF), we have transfected the SH-SY5Y neuroblastoma cell line, which expresses mu-and delta-opioid receptors, with the human NPFF2receptor. The selected clone, SH2-D9, expressed high-affinity NPFF2 receptors in the same range order as mu- and delta-opioid receptors (100-300 fmol/mg of protein). The NPFF analog [D-Tyr1, (NMe)Phe3]NPFF (1DMe) did not modify the binding parameters of the mu- and delta-specific agonists [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin and deltorphin-I, respectively. 1DMe dose dependently inhibited 75 to 80% of the cAMP production stimulated by forskolin. Preincubation with 1DMe halved the maximal inhibition of N-type Ca2+ channels by opioid agonists. In the presence of carbachol, acting on muscarinic receptors to release Ca2+ from the intracellular stores, deltorphin-I and 1DMe enhanced this release. Preincubation with 1DMe reduced the maximal effect of deltorphin-I by 40%, demonstrating an antiopioid effect in this experimental model for the first time. By using peptides corresponding to the carboxyl terminus of the alphai1,2, alphai3, alphao, and alphas subunits of G proteins, which specifically uncouple receptors from G proteins, we demonstrated that mu-opioid and NPFF2 receptors couple to the four subunits assayed. The Ca2+ release from the intracellular stores by 1DMe resulted from the coupling of NPFF2 receptors with Galphao and Galphai1,2, whereas the coupling with Galphas reduced the antiopioid effect of 1DMe in the modulation of N-type channels. This SH2-D9 cell line now provides the opportunity to study the interaction between both receptors.
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Affiliation(s)
- Catherine Mollereau
- Institut de Pharmacologie et de Biologie Structurale, CNRS, UMR 5089, 205 route de Narbonne, 31077 Toulouse cedex 04, France
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Bonnard E, Burlet-Schiltz O, Monsarrat B, Girard JP, Zajac JM. Identification of proNeuropeptide FFA peptides processed in neuronal and non-neuronal cells and in nervous tissue. ACTA ACUST UNITED AC 2003; 270:4187-99. [PMID: 14519131 DOI: 10.1046/j.1432-1033.2003.03816.x] [Citation(s) in RCA: 20] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
Peptides which should be generated from the neuropeptide FF (NPFF) precursor were identified in a neuronal (human neuroblastoma SH-SY5Y) cell line and in COS-7 cells after transient transfection of the human proNPFFA cDNA and were compared with those detected in the mouse spinal cord. After reverse-phase high performance liquid chromatography of soluble material, NPFF-related peptides were immunodetected with antisera raised against NPFF and identified by using on-line capillary liquid chromatography/nanospray ion trap tandem mass spectrometry. Neuronal and non-neuronal cells generated different peptides from the same precursor. In addition to NPFF, SQA-NPFF (Ser-Gln-Ala-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-amide) and NPAF were identified in the human neuroblastoma while only NPFF was clearly identified in COS-7 cells. In mouse, in addition to previously detected NPFF and NPSF, SPA-NPFF (Ser-Pro-Ala-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-amide), the homologous peptide of SQA-NPFF, were characterized. These data on intracellular processing of proNeuropeptide FFA are discussed in regard to the known enzymatic processing mechanisms.
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Affiliation(s)
- Elisabeth Bonnard
- Institut de Pharmacologie et de Biologie Structurale, Toulouse, France
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Li LS, Qu RY, Wang W, Guo H. Significance of changes of gastrointestinal peptides in blood and ileum of experimental spleen deficiency rats. World J Gastroenterol 2003; 9:553-6. [PMID: 12632517 PMCID: PMC4621581 DOI: 10.3748/wjg.v9.i3.553] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To explore the mechanism of spleen deficiency (SD) by studying the relationship of gastro-intestinal peptides level and ileal electro-mechanical activity of SD rats and cold restrain rats.
METHODS: (1) spleen deficiency (SD) model was established by feeding Houpou:Zhishi: Dahuang in the ratio of 3:3:2, 3 ml/time, for 42 days. (2) The cold restrain stress model: Animals were restrained on grille and placed in a cool water at 18 °C for 3 h. (3) Substance P (SP) and vasoactive intestinal peptide (VIP) levels in all layers of initial part of ileum and blood in rats were measured by radioimmunoassays (RIA) while changes of electric activity and motility in ileum of rats were recorded with electrode and strain gauge.
RESULTS: SP levels in ileum and blood of experimental SD rats were significantly higher than that of the control groups (9.89 ± 5.65 vs 1.22 ± 1.18, P < 0.005, in ileum; 22.7 ± 3.95 vs 6.60 ± 1.47, P < 0.001, in blood) while the VIP levels of the SD rats were significantly lower than that of the controls (3.50 ± 2.01 vs 9.10 ± 4.91, P < 0.05, in ileum; 229.8 ± 62.4 vs 560.4 ± 151.3, P < 0.001, in blood). As compared with the controls, the average frequency of slow electric waves (21.3 ± 0.96 vs 18.2 ± 2.28, P < 0.05) and motility (21.5 ± 0.58 vs 18 ± 2.65, P < 0.005) of SD rats increased obviously and the frequency of fast waves of SD rat also increased. In spontaneous recovery cases, SP levels recovered significantly (compared with the SD groups, 2.99 ± 0.62 vs 9.89 ± 5.65, P < 0.001, in ileum; 14.4 ± 4.22 vs 22.7 ± 3.95, P < 0.001, in blood) but did not drop to normal. After the SD rats treated with Chinese herbs (Jiawei Sijun zi Tang), SP improved (compared with SD cases, 2.20 ± 1.25 vs 9.89 ± 5.65, (P < 0.001), in ileum; 10.7 ± 1.88 vs 22.7 ± 3.95, (P < 0.001), in blood) and VIP in blood also improved (compared with SD rats, 485.7 ± 229.0 vs 229.8 ± 62.4, P < 0.01) while the amplitude of motility decreased apparently (compared with the SD rats, 0.64 ± 0.096 vs 0.89 ± 0.15, P < 0.01). The ileal SP levels of cool stress didn’t change while the ileal VIP levels of cool stress became significantly lower than that of the control groups (2.87 ± 0.87 vs 9.10 ± 4.91, P < 0.01). The blood SP levels of cool stress were significantly higher (15.60 ± 1.83 vs 6.60 ± 1.47, P < 0.001) whereas the blood VIP levels of cool stress were significantly lower than that of the control group (153.4 ± 70.46 vs 560.4 ± 151.30, P < 0.001).
CONCLUSION: Changes of SP and VIP levels in initial part of ileum and blood of SD rats and cool stress rats may be closely related to the gastrointestinal motility disorders presented in SD and cool stress rats. the Chinese herbs (Jiawei Sijun zi Tang) currently used have partially therapeutic effect.
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Affiliation(s)
- Li-Sheng Li
- Department of Physiology, Capital University of Medical Sciences, 100054, Beijing, China.
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