TY - THES A1 - Kloka, Jan Andreas T1 - Endogene Lipide als neues Behandlungstarget im TRPA1-vermittelten Entzündungsschmerz T1 - Endogenous lipids as a new treatment target in TRPA1-mediated inflammatory pain N2 - For nociceptive wound pain, the transient receptor potential channel(TRP) mediated calcium influx is essential. Reactive oxygen species (ROS) and their downstream oxidation products such as 4-hydroxynonenal activate the ankyrin 1 homologue TRPA1 in vivo and in vitro. The calcium imaging experiments performed in this study were carried out on stably with TRPA1 and TRPV1 transfected HEK-293 cells and spinal dorsal root ganglion neurons to further understand the mechanistic correlations of nociceptive pain development in inflammatory wound pain. E06, a monoclonal autoantibody (mAb) against oxidized phosphatidylcholine (OxPC) and D-4F, a mimetic peptide of the structural protein apolipoprotein A-I of high density lipoproteins (HDL) were previously used as a diagnostic tools and novel compounds in atherosclerosis. In this study, E06 mAb and D-4F peptide, both, reduced the TRPA1-mediated calcium influx in vitro caused by lipid peroxidation products (OxPL) such as 4-HNE and reactive oxygen species such as H2O2. In addition, we discovered that neither E06 mAb nor D-4F showed a calcium influx-relevant interaction with the Transient Receptor Potential Channel Vanillin 1 (TRPV1) activator capsaicin or the TRPV1 channel itself. Taken together, E06 mAb and D-4F peptide are two promising substances to reduce inflammatory pain and local pain relief. N2 - Für nozizeptive Wundschmerzen ist der Transient Receptor Potential Channel (TRP) vermittelte Kalziumeinstrom unerlässlich. Reaktive Sauerstoffspezies (ROS) und deren Oxidationsprodukte wie 4-Hydroxynonenal (4-HNE) aktivieren das Ankyrin-1-Homolog TRPA1 in vivo und in vitro. Die in dieser Studie durchgeführten Kalzium-Imaging Experimente wurden an stabil mit TRPA1 und TRPV1 transfizierten HEK-293-Zellen und spinalen Hinterwurzelganglien durchgeführt, um die mechanistischen Zusammenhänge der nozizeptiven Schmerzentstehung bei entzündlichen Wundschmerzen besser zu verstehen. E06, ein monoklonaler Autoantikörper (mAb) gegen oxidiertes Phosphatidylcholin (OxPC) und D-4F, ein mimetisches Peptid des Strukturproteins Apolipoprotein A-I aus dem high density lipoprotein (HDL), wurden bisher als diagnostischer Marker bei Atherosklerose eingesetzt. In den durchgeführten Experimenten reduzierten E06 mAb und D-4F den durch Lipidperoxidationsprodukte (OxPL) wie 4-HNE und reaktive Sauerstoffspezies wie H2O2 verursachten TRPA1-vermittelten Kalziumeinfluss in vitro. Darüber hinaus zeigte sich, dass weder E06 mAb noch D-4F eine Kalziumeinstromrelevante Interaktion mit dem Transient Receptor Potential Channel Vanillin 1 (TRPV1)-Aktivator Capsaicin oder dem TRPV1-Kanal aufweisen. E06 mAb und ApoA-I mimetisches Peptid D-4F erscheinen deshalb als zwei vielversprechende Substanzen, um den inflammatorischen Wundschmerz zu verringern. Deshalb sind sie auch als potentielles Analgetikum für eine nebenwirkungsärmere, lokale Schmerzbekämpfung vielversprechend. KW - Entzündung KW - Schmerztherapie KW - Schmerzforschung KW - TRPA1 KW - H2O2 KW - Calcium Imaging KW - 4-HNE KW - ApoA-I D-4F KW - E06 mAb KW - Phospholipide KW - Ionenkanal Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-180844 ER - TY - JOUR A1 - Oehler, Beatrice A1 - Kloka, Jan A1 - Mohammadi, Milad A1 - Ben-Kraiem, Adel A1 - Rittner, Heike L. T1 - D-4F, an ApoA-I mimetic peptide ameliorating TRPA1-mediated nocifensive behaviour in a model of neurogenic inflammation JF - Molecular Pain N2 - Background High doses of capsaicin are recommended for the treatment of neuropathic pain. However, low doses evoke mechanical hypersensitivity. Activation of the capsaicin chemosensor transient receptor potential vanilloid 1 (TRPV1) induces neurogenic inflammation. In addition to the release of pro-inflammatory mediators, reactive oxygen species are produced. These highly reactive molecules generate oxidised phospholipids and 4-hydroxynonenal (4-HNE) which then directly activate TRP ankyrin 1 (TRPA1). The apolipoprotein A-I mimetic peptide D-4F neutralises oxidised phospholipids. Here, we asked whether D-4F ameliorates neurogenic hypersensitivity in rodents by targeting reactive oxygen species and 4-HNE in the capsaicin-evoked pain model. Results Co-application of D-4F ameliorated capsaicin-induced mechanical hypersensitivity and allodynia as well as persistent heat hypersensitivity measured by Randell–Selitto, von Frey and Hargreaves test, respectively. In addition, mechanical hypersensitivity was blocked after co-injection of D-4F with the reactive oxygen species analogue H2O2 or 4-HNE. In vitro studies on dorsal root ganglion neurons and stably transfected cell lines revealed a TRPA1-dependent inhibition of the calcium influx when agonists were pre-incubated with D-4F. The capsaicin-induced calcium influx in TRPV1-expressing cell lines and dorsal root ganglion neurons sustained in the presence of D-4F. Conclusions D-4F is a promising compound to ameliorate TRPA1-dependent hypersensitivity during neurogenic inflammation. KW - TRPA1 KW - capsaicin KW - reactive oxygen species KW - oxidised lipids KW - pain KW - targeting Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-236061 VL - 16 ER - TY - JOUR A1 - Oehler, Beatrice A1 - Kistner, Katrin A1 - Martin, Corinna A1 - Schiller, Jürgen A1 - Mayer, Rafaela A1 - Mohammadi, Milad A1 - Sauer, Reine-Solange A1 - Filipovic, Milos R. A1 - Nieto, Francisco R. A1 - Kloka, Jan A1 - Pflücke, Diana A1 - Hill, Kerstin A1 - Schaefer, Michael A1 - Malcangio, Marzia A1 - Reeh, Peter W. A1 - Brack, Alexander A1 - Blum, Robert A1 - Rittner, Heike L. T1 - Inflammatory pain control by blocking oxidized phospholipid-mediated TRP channel activation JF - Scientific Reports N2 - Phospholipids occurring in cell membranes and lipoproteins are converted into oxidized phospholipids (OxPL) by oxidative stress promoting atherosclerotic plaque formation. Here, OxPL were characterized as novel targets in acute and chronic inflammatory pain. Oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (OxPAPC) and its derivatives were identified in inflamed tissue by mass spectrometry and binding assays. They elicited calcium influx, hyperalgesia and induced pro-nociceptive peptide release. Genetic, pharmacological and mass spectrometric evidence in vivo as well as in vitro confirmed the role of transient receptor potential channels (TRPA1 and TRPV1) as OxPAPC targets. Treatment with the monoclonal antibody E06 or with apolipoprotein A-I mimetic peptide D-4F, capturing OxPAPC in atherosclerosis, prevented inflammatory hyperalgesia, and in vitro TRPA1 activation. Administration of D-4F or E06 to rats profoundly ameliorated mechanical hyperalgesia and inflammation in collagen-induced arthritis. These data reveal a clinically relevant role for OxPAPC in inflammation offering therapy for acute and chronic inflammatory pain treatment by scavenging OxPAPC. KW - chronic pain KW - ion channels in the nervous system KW - molecular medicine KW - pain Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-158536 VL - 7 IS - 5447 ER -