TY - JOUR A1 - Üçeyler, Nurcan A1 - Valet, Michael A1 - Kafke, Waldemar A1 - Tölle, Thomas R. A1 - Sommer, Claudia T1 - Local and Systemic Cytokine Expression in Patients with Postherpetic Neuralgia N2 - Background Postherpetic neuralgia (PHN) is the painful complication of a varicella zoster virus reactivation. We investigated the systemic and local gene expression of pro- and anti-inflammatory cytokine expression in patients with PHN. Methods Thirteen patients with PHN at the torso (Th4-S1) were recruited. Skin punch biopsies were obtained from the painful and the contralateral painless body area for intraepidermal nerve fiber density (IENFD) and cytokine profiling. Additionally, blood was withdrawn for systemic cytokine expression and compared to blood values of healthy controls. We analyzed the gene expression of selected pro- and anti-inflammatory cytokines (tumor necrosis factor-alpha [TNF] and interleukins [IL]-1β, IL-2, and IL-8). Results IENFD was lower in affected skin compared to unaffected skin (p<0.05), while local gene expression of pro- and anti-inflammatory cytokines did not differ except for two patients who had 7fold higher IL-6 and 10fold higher IL-10 gene expression in the affected skin compared to the contralateral unaffected skin sample. Also, the systemic expression of cytokines in patients with PHN and in healthy controls was similar. Conclusion While the systemic and local expression of the investigated pro- and anti-inflammatory cytokines was not different from controls, this may have been influenced by study limitations like the low number of patients and different disease durations. Furthermore, other cytokines or pain mediators need to be considered. KW - neuropathic pain KW - cytokines KW - pain sensation KW - gene expression KW - nerve fibres KW - RNA extraction KW - shingles KW - skin tumors Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-113041 ER - TY - THES A1 - Wang, Ying T1 - Immune and peripheral endogenous opioid mechanisms of electroacupuncture analgesia T1 - Immunologische und periphere endogene Opioidmechanismen bei Analgesie durch Elektroakupunktur N2 - A precious treasure in traditional Chinese medicine (TCM), acupuncture played a vital and irreplaceable role in contributing to people’s health in the thousands of years of Chinese history, and in 2010 was officially added to the “Representative List of the Intangible Cultural Heritage of Humanity” by the United Nations. Because of the side-effects of long-term drug therapy for pain, and the risks of dependency, acupuncture has been widely accepted as one of the most important alternative choice therapies for treating varieties of acute and chronic pain-related disorders. The clinical application and scientific mechanism research of acupuncture have therefore increased intensively in the last few decades. Besides hand acupuncture, other treatment approaches e.g. electroacupuncture (EA) have been widely accepted and applied as an important acupuncture-related technique for acupuncture analgesia (AA) research. The involvement of opioid peptides and receptors in acute AA has been shown via pre-EA application of opioid receptor/peptide antagonists. However, existing publications still cannot illuminate the answer to the following question: how does sustained antinociception happen by EA treatment? The hypothesis of opioid peptide-mediated tonic AA might be able to answer the question. In the first part of this thesis, the institution of a reproducible acupuncture treatment model as well as the endogenous opioid-related mechanisms was demonstrated. An anatomically-based three-dimensional (3D) rat model was established to exhibit a digital true-to-life organism, accurate acupoint position and EA treatment protocol on bilateral acupoint GB-30 Huantiao. The optimal EA treatment protocol (100 Hz, 2-3 mA, 0.1 ms, 20 min) at 0 and 24 h after induction of inflammatory pain by complete Freund’s adjuvant (CFA) on conscious free-moving rats was then established. EA elicited significant sustained mechanical and thermal antinociception up to 144 h. Post-EA application of opioid receptors (mu opioid receptor, MOR; delta opioid receptor, DOR) antagonists naloxone (NLX) and naltrindole (NTI), or opioid peptide antibodies anti-beta-endorphin (anti-END), met-enkephalin (anti-ENK) or -dynorphin A (anti-DYN) could also block this effect at a late phase (96 h) of CFA post-EA, which suggested opioid-dependent tonic analgesia was produced by EA. Meanwhile, EA also reduced paw temperature and volume at 72-144 h post CFA indicating anti-inflammatory effects. Nociceptive thresholds were assessed by paw pressure threshold (Randall-Sellito) or paw withdrawal latency (Hargreaves) and an anti-inflammatory effect was evaluated by measurement of plantar temperature and volume of inflamed paw. The second part of the thesis further suggests the correlation between the chemokine CXCL10 (= interferon-gamma inducible protein 10, IP-10) and opioid peptides in EA-induced antinociception. Based on a comprehensive Cytokine Array of 29 cytokines, targeted cytokines interleukin (IL)-1alpha, interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, interleukin (IL)-4, interleukin (IL)-13, interferon (IFN)-gamma as well as CXCL10 were selected and quantified by enzyme-linked immunosorbent assay (ELISA), and real time reverse transcription-polymerase chain reaction (RT-PCR) quantification confirmed upregulation of CXCL10 mRNA at both 72 and 96 h. The following hyperalgesic assessment suggested the antinociceptive effect of CXCL10. The double immunostaining localizing opioid peptides with macrophages expressed the evident upregulation of CXCR3-receptor of CXCL10 in EA treated samples as well as the significant upregulation or downregulation of opioid peptides by repeated treatment of CXCL10 or antibody of CXCL10 via behavioral tests and immune staining. Subsequent immunoblotting measurements showed non-alteration of opioid receptor level by EA, indicating that the opioid receptors did not apparently contribute to AA in the present studies. In vitro, CXCL10 did not directly trigger opioid peptide END release from freshly isolated rat macrophages. This might implicate an indirect property of CXCL10 in vitro stimulating the opioid peptide-containing macrophages by requiring additional mediators in inflammatory tissue. In summary, this project intended to explore the peripheral opioid-dependent analgesic mechanisms of acupuncture with a novel 3D treatment rat model and put forward new information to support the pivot role of chemokine CXCL10 in mediating EA-induced tonic antinociception via peripheral opioid peptides. N2 - Als wertvoller Schatz in der traditionellen chinesischen Medizin (TCM) spielt die Akupunktur eine wichtige und unersetzliche Rolle für die Gesundheit der Menschen in der über tausendjährigen Geschichte von China und wurde im Jahr 2010 offiziell in das "Weltkulturerbe" der Vereinten Nationen aufgenommen. Aufgrund der Nebenwirkungen von Langzeittherapien zur Schmerzbehandlung und dem Risiko der Abhängigkeit wird Akupunktur weithin als eine wichtigste Alternative für die Behandlung von akuten und chronischen Schmerzen eingesetzt. Die klinische Anwendung und Forschung in der Akupunktur wurden in den letzten Jahrzehnten intensiv vorangetrieben. Neben Handakupunktur gibt es noch andere Behandlungsmöglichkeiten, wie z.B. die Elektroakupunktur (EA). EA ist vor allem eine allgemein akzeptierte und wichtige akupunkturbezogene Technik für die Akupunkturanalgesie (AA) in der Forschung. Die Beteiligung von Opioidpeptiden und Opioidrezeptoren in der akuten AA wurde mittels Anwendung von Opioidrezeptorantagonisten/Opioidpeptidantikörpern appliziert vor EA gezeigt. Nach dem aktuellen Forschungsstand kann man jedoch nicht die Frage beantworten, wie die längerfristige (tonische) Antinozizeption nach EA-Behandlung funktioniert. Mit einer Hypothese zur Opioidpeptid vermittelten tonischen AA könnte man die Frage hierzu beantworten. In der vorliegenden Arbeit wurden in einem Modell der Akupunktur zum ersten Mal endogene Opioid-vermittelte Mechanismen reproduzierbar nachgewiesen. Es wurde ein dreidimensionales (3D) anatomisch-basiertes Rattenmodell entworfen, um am wachen Tier ein genaue Akupunkturbehandlung (EA) an den Akupunkten GB-30 Huantiao beidseitig durchzuführen. Darüber hinaus wurde ein optimiertes Behandlungsprotokoll von EA (100 Hz, 2-3 mA, 0.1 ms, 20 min) bei Ratten 0 und 24 h nach intraplantarer Injektion von komplettem Freunds Adjuvans (CFA) etabliert. Nozizeptive Schwellen wurden mittels Pfotendruckschwelle (Randall-Sellito) oder Pfotenrückzugslatenzzeit (Hargreaves) gemessen und die entzündungshemmende Wirkung durch Messung der Pfotentemperatur und Volumen der entzündeten Pfote ausgewertet. EA bewirkte eine signifikante mechanische und thermische Antinozizeption, welche bis zu 144 h anhielt. Die antinozizeptive Wirkung durch EA war nach Injektion von Opioidrezeptorantagonisten (mu opioid receptor, MOR; delta opioid receptor, DOR) Naloxon (NLX) und Naltrindol (NTI) oder Antikörpern gegen die Opioidpeptide beta-Endorphin (anti-END), Met-Enkephalin (anti-ENK) oder Dynorphin A (anti-DYN) während der späten Entzündungsphase (96 h) mit CFA blockierbar. Dies lässt auf eine durch EA induzierte tonische Analgesie schließen. Darüber hinaus reduzierte EA auch die erhöhte Pfotentemperatur und das erhöhte Pfotenvolumen, welche sehr typisch ist für eine 96-144 h Entzündung mit CFA. Dies spricht für eine entzündungshemmende Wirkung von EA. Nachfolgend wurde die Beteiligung von Opioidpeptiden und dem Chemokin CXCL10 (= Interferon-gamma induziertes Protein 10, IP-10) sowie die Korrelationen zwischen beiden geklärt. Basierend auf umfangreichenden Zytokinarrays mit 29 Zytokinen, wurden die Zytokine Interleukin (IL)-1alpha, Interleukin (IL)-1beta, Tumornekrosefaktor (TNF)-alpha, Interleukin (IL)-4, Interleukin (IL)-13, Interferon (IFN)-gamma und CXCL10 gezielt ausgewählt und im Enzym Immunoassay (ELISA) sowie durch eine Echtzeit Reverse Transkription-Polymerase Kettenreaktion (RT-PCR) quantifiziert. Die Quantifizierung auf mRNA-Ebene zeigte eine Hochregulation von CXCL10 bei 96 h und zum früheren Zeitpunkt von 72 h. Nachfolgende Schmerzschwellenmessungen wiesen auf eine antinozizeptive Wirkung von CXCL10 hin. Eine Doppelimmunfärbung zeigte die Lokalisation von Opioidpeptiden in Makrophagen. Nachweislich wurde die Expression des CXCR3-Rezeptor von CXCL10 in EA behandelten Pfoten erhöht. Ebenso kam es zu einer signifikanten Hochregulation von Opioidpeptiden durch wiederholte Behandlung mit CXCL10 bzw. Herunterregulation der Opioidpeptide nach wiederholter Behandlung mit anti-CXCL10 Antikörper. Dies wurde sowohl in Verhaltenstests als auch in der Immunfärbung zu beobachtet. Immunoblotting zeigte keine Veränderung der Expression von Opioidrezeptoren nach EA, was schließen lässt, dass die Menge an Opioidrezeptoren in den vorliegenden Untersuchungen keine Rolle spielen. Da CXCL10 in vitro keine direkten Effekt auf die Freisetzung des Opioidpeptides beta-END aus frisch isolierten Rattenmakrophagen hat, liegt die Vermutung nahe, dass CXCL10 in vitro eine indirekte Rolle als Mediator zukommt und indirekt die Opioidpeptidfreisetzung aus Makrophagen in entzündlichen Gewebe stimuliert wird. Zusammenfassend wurden in dem hier vorgestellten Projekt die Opioidpeptid-abhängigen analgetischen und peripheren Mechanismen der Akupunktur mit einem neuartigem 3D-Behandlungsmodell der Ratte untersucht und eine Schlüsselrolle des Chemokins CXCL10 bei der Vermittlung der EA-induzierten tonischen Antinozizeption in Abhändigkeit von peripheren Opioidpeptiden. (Translated by Dr. Dagmar Hackel and revised by Priv.-Doz. Dr. Heike Rittner) KW - Elektroakupunktur KW - electroacupuncture KW - peripheral analgesia KW - Chemokin CXCL10 KW - Analgesie KW - Cytokine KW - Ratte KW - opioid peptide KW - cytokines KW - Periphere Analgesie KW - Opioidpeptide Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-98326 ER - TY - JOUR A1 - Rittner, Heike L. A1 - Wang, Ying A1 - Gehringer, Rebekka A1 - Mousa, Shaaban A. A1 - Hackel, Dagmar A1 - Brack, Alexander T1 - CXCL10 Controls Inflammatory Pain via Opioid Peptide- Containing Macrophages in Electroacupuncture N2 - Acupuncture is widely used for pain treatment in patients with osteoarthritis or low back pain, but molecular mechanisms remain largely enigmatic. In the early phase of inflammation neutrophilic chemokines direct opioid-containing neutrophils in the inflamed tissue and stimulate opioid peptide release and antinociception. In this study the molecular pathway and neuroimmune connections in complete Freund's adjuvant (CFA)-induced hind paw inflammation and electroacupuncture for peripheral pain control were analyzed. Free moving Wistar rats with hind paw inflammation were treated twice with electroacupuncture at GB30 (Huan Tiao - gall bladder meridian) (day 0 and 1) and analyzed for mechanical and thermal nociceptive thresholds. The cytokine profiles as well as the expression of opioid peptides were quantified in the inflamed paw. Electroacupuncture elicited long-term antinociception blocked by local injection of anti-opioid peptide antibodies (beta-endorphin, met-enkephalin, dynorphin A). The treatment altered the cytokine profile towards an anti-inflammatory pattern but augmented interferon (IFN)-gamma and the chemokine CXCL10 (IP-10: interferon gamma-inducible protein) protein and mRNA expression with concomitant increased numbers of opioid peptide-containing CXCR3+ macrophages. In rats with CFA hind paw inflammation without acupuncture repeated injection of CXCL10 triggered opioid-mediated antinociception and increase opioid-containing macrophages. Conversely, neutralization of CXCL10 time-dependently decreased electroacupuncture-induced antinociception and the number of infiltrating opioid peptide-expressing CXCR3+ macrophages. In summary, we describe a novel function of the chemokine CXCL10 - as a regulator for an increase of opioid-containing macrophages and antinociceptive mediator in inflammatory pain and as a key chemokine regulated by electroacupuncture. KW - opioids KW - inflammation KW - macrophages KW - cytokines KW - chemokines KW - enzyme-linkes immunoassays KW - acupuncture KW - analysis of variance Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-112979 ER - TY - JOUR A1 - Meyer zu Hörste, Gerd A1 - Cordes, Steffen A1 - Mausberg, Anne K. A1 - Zozulya, Alla L. A1 - Wessig, Carsten A1 - Sparwasser, Tim A1 - Mathys, Christian A1 - Wiendl, Heinz A1 - Hartung, Hans-Peter A1 - Kieseier, Bernd C. T1 - FoxP3+Regulatory T Cells Determine Disease Severity in Rodent Models of Inflammatory Neuropathies JF - PLOS ONE N2 - Inflammatory neuropathies represent disabling human autoimmune disorders with considerable disease variability. Animal models provide insights into defined aspects of their disease pathogenesis. Forkhead box P3 (FoxP3)+ regulatory T lymphocytes (Treg) are anti-inflammatory cells that maintain immune tolerance and counteract tissue damage in a variety of immune-mediated disorders. Dysfunction or a reduced frequency of Tregs have been associated with different human autoimmune disorders. We here analyzed the functional relevance of Tregs in determining disease manifestation and severity in murine models of autoimmune neuropathies. We took advantage of the DEREG mouse system allowing depletion of Treg with high specificity as well as anti-CD25 directed antibodies to deplete Tregs in mice in actively induced experimental autoimmune neuritis (EAN). Furthermore antibody-depletion was performed in an adoptive transfer model of chronic neuritis. Early Treg depletion increased clinical EAN severity both in active and adoptive transfer chronic neuritis. This was accompanied by increased proliferation of myelin specific T cells and histological signs of peripheral nerve inflammation. Late stage Treg depletion after initial disease manifestation however did not exacerbate inflammatory neuropathy symptoms further. We conclude that Tregs determine disease severity in experimental autoimmune neuropathies during the initial priming phase, but have no major disease modifying function after disease manifestation. Potential future therapeutic approaches targeting Tregs should thus be performed early in inflammatory neuropathies. KW - Guillain-Barre-Syndrome KW - regulatory cells KW - C57BL/6 mice KW - demyelinating polyradiculoneuropathy KW - cytokines KW - pathogenesis KW - polyneuropathy KW - enteropathy KW - peptide KW - experimental autoimmune neuritis Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-115239 VL - 9 IS - 10 ER - TY - JOUR A1 - Enigk, Fabian A1 - Wagner, Antje A1 - Samapati, Rudi A1 - Rittner, Heike A1 - Brack, Alexander A1 - Mousa, Shaaban A. A1 - Schäfer, Michael A1 - Habazettl, Helmut A1 - Schäper, Jörn T1 - Thoracic epidural anesthesia decreases endotoxin-induced endothelial injury JF - BMC Anesthesiology N2 - Background: The sympathetic nervous system is considered to modulate the endotoxin-induced activation of immune cells. Here we investigate whether thoracic epidural anesthesia with its regional symapathetic blocking effect alters endotoxin-induced leukocyte-endothelium activation and interaction with subsequent endothelial injury. Methods: Sprague Dawley rats were anesthetized, cannulated and hemodynamically monitored. E. coli lipopolysaccharide (Serotype 0127: B8, 1.5 mg x kg(-1) x h(-1)) or isotonic saline (controls) was infused for 300 minutes. An epidural catheter was inserted for continuous application of lidocaine or normal saline in endotoxemic animals and saline in controls. After 300 minutes we measured catecholamine and cytokine plasma concentrations, adhesion molecule expression, leukocyte adhesion, and intestinal tissue edema. Results: In endotoxemic animals with epidural saline, LPS significantly increased the interleukin-1 beta plasma concentration (48%), the expression of endothelial adhesion molecules E-selectin (34%) and ICAM-1 (42%), and the number of adherent leukocytes (40%) with an increase in intestinal myeloperoxidase activity (26%) and tissue edema (75%) when compared to healthy controls. In endotoxemic animals with epidural infusion of lidocaine the values were similar to those in control animals, while epinephrine plasma concentration was 32% lower compared to endotoxemic animals with epidural saline. Conclusions: Thoracic epidural anesthesia attenuated the endotoxin-induced increase of IL-1 beta concentration, adhesion molecule expression and leukocyte-adhesion with subsequent endothelial injury. A potential mechanism is the reduction in the plasma concentration of epinephrine. KW - endotoxemia KW - myeloperoxidase KW - endothelial injury KW - adhesion molecules KW - inflammatory response KW - intestinal microvascular perfusion KW - cell-adhesion KW - induced impairment KW - reperfusion injury KW - sepsis KW - neutrophil KW - lidocaine KW - lung injury KW - cytokines KW - epidural anesthesia Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-116787 VL - 14 IS - 23 ER -