610 Medizin und Gesundheit
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Pericytes are capillary-associated mural cells involved in the maintenance and the stability of the vascular network. This thesis aims to investigate the role of pericytes in the heart in the context of ageing and disease. We highlight the malignant effects of the remodelling in the heart and stress the focus on the role of cardiac pericytes in this context. We show that ageing reduces pericyte coverage and that myocardial infarction (MI) causes an activation of these cells. Single-nuclei and single-cell RNA sequencing analysis of murine hearts further revealed that the expression of the Regulator of G-protein signalling 5 (Rgs5) is reduced in cardiac pericytes both in ageing and transiently at day 1 and day 3 after MI. The loss of RGS5 in pericytes drives an entropic state of these mural cells characterized by morphological changes, excessive extracellular deposition and enhanced Gaq mediated GPCR signalling. The deletion of RGS5 in pericytes causes cardiac systolic dysfunction, induces myocardial fibrosis, and drives the activation of cardiac fibroblasts in a TGFb-dependent manner. In conclusion, our results describe the importance of pericytes maintaining cardiac homeostasis, identify RGS5 as a key regulator of this process and propose pericytes as crucial mediators of cardiac fibrosis and possible therapeutic targets to prevent cardiovascular disease.
Die Ergebnisse der Studie und die Diversität der Datenbanken ist groß.
Für 12 Datenbanken wurde ein Punktesystem mit elf Items entworfen, um die Qualität der einzelnen Datenbanken zu objektivieren. Keine Datenbank konnte alle Bewertungskriterien erfüllen. Der insgesamt schlechte Punktedurchschnitt ist ein Indikator für die Mängel der aktuell verfügbaren Datenbanken. Außerdem konnten wir einen Qualitätsunterschied zwischen kostenpflichtigen und kostenfreien Datenbanken beweisen und mussten im Zuge dieser Ergebnisse die Frage stellen, ob kostenfreie Datenbanken überhaupt nützlich sind. Zwischen den kostenpflichtigen Datenbanken fallen die Qualitätsunterschiede weniger gravierend aus, wenngleich Stärken und Schwächen sich deutlich unterscheiden. Die häufigsten Wechselwirkungen wurden in allen Datenbanken mit großem Abstand zwischen rein psychiatrischen Interaktionspaaren erfasst. Dieses zeigt, wie wechselwirkungsreich Psychopharmaka sind und dass psychiatrische Patienten besonders vulnerabel sind. Die Nutzung digitaler Hilfsmittel scheint bei Betrachtung der hohen Anzahl ausgegebener Warnmeldungen unabdingbar zu sein, dennoch existiert große Uneinheitlichkeit bei der Bewertung der einzelnen Interaktionen. Die Vorstellung, dass zwei Kliniker bei Nutzung zweier unterschiedlicher Datenbanken zu völlig unterschiedlichen Empfehlungen kommen, fällt nicht schwer. Gleichzeitig könnte die Kooperation von Heilberuflern, die unterschiedliche Datenbanken verwenden, die Chance auf zusätzlichen Informationsgewinn und Austausch erhöhen, was im Umkehrschluss in einer erhöhten Arzneimitteltherapiesicherheit resultiert. In Studien konnte der positive Effekt interdisziplinärer Zusammenarbeit bereits bewiesen werden.
Zusammenfassend konnten umfangreiche Differenzen zwischen allen untersuchten Datenbanken aufgezeigt werden. Um den Anforderungen des klinischen Alltags zu genügen, müssen digitale Unterstützungssysteme weiterentwickelt werden.
Die „ideale Datenbank“ gibt es bisher nicht – das lässt sich durch unser Punktesystem beweisen. Um im klinischen Alltag Patientensicherheit zu gewährleisten ist die Nutzung einer einzelnen Datenbank bisher nicht ausreichend.
Die Gewährung der Patientensicherheit sollte unser oberstes Ziel sein und um dieses zu erreichen, bedarf es vieler Komponenten. Neben der Nutzung und vor allem Weiterentwicklung digitaler Unterstützungssysteme sollte auch der zwischenmenschliche Austausch weiter gefördert werden. Interdisziplinäre Zusammenarbeit im Sinne pharmazeutischer Dienstleistungen zur Medikationsanalyse könnten ein zusätzliches Instrument zur Vermeidung arzneimittelbezogener Probleme werden.
Zukünftig werden unsere Patienten am meisten von optimaler Nutzung weiterentwickelter Technologien, sowie wachsendem zwischenmenschlichem Austausch profitieren.
Multilevel-Untersuchung des nitrinergen Systems bei affektiven Störungen und schizophrenen Psychosen
(2023)
Das nitrinerge System und damit auch NOx als Neurotransmitter werden mit der Entstehung verschiedener psychischer Erkrankungen in Verbindung gebracht. Die genaue Rolle des Botenstoffs ist jedoch nicht ausreichend geklärt und auch die Frage, ob dieser als diagnostischer oder prädiktiver Biomarker nützlich sein könnte, ist unbeantwortet. In der vorliegenden Arbeit wurde folglich untersucht, ob es Unterschiede zwischen den Diagnosegruppen MDD, BIP, SCZ und der Kontrollgruppe bezüglich peripherer NOx- Konzentrationen gibt. Darüber hinaus wurden Unterschiede innerhalb der Diagnosegruppen im Krankheitsverlauf im Sinne von Phasenunterschieden mittels zweier Messzeitpunkte untersucht und analysiert, ob es Korrelationen mit genetischen Variationen in NOS-Genabschnitten gibt. Insgesamt wurden 185 Probanden in die Studie mitaufgenommen: 52 gesunde CTRL, 43 Patienten mit MDD, 41 Patienten mit BIP und 49 Patienten mit SCZ. Biochemische, genetische und klinische Daten wurden bei Aufnahme und Entlassung in der psychiatrischen Abteilung des Universitätsklinikums Frankfurt erhoben. Klinische Daten, die den Symptomverlauf 90 und die Erkrankungsschwere beurteilten, nutzten dazu standardisierte teilstrukturierte klinische Interviews. Biochemische Daten wurden mittels im Serum gemessener NOx- Spiegel quantifiziert. Bezüglich der Untersuchung der Risikogenvarianten wurden Probanden anhand des NOS1 ex1f-VNTR-Polymorphismus sowie SNPs in den Genen NOS1, NOS3 und NOS1AP genotypisiert. Bei Aufnahme wiesen SCZ-Patienten im Vergleich zu CTRL-, MDD- und BIP-Gruppen signifikant höhere NOx- Konzentrationen auf. Während NOx- Spiegel im Behandlungsverlauf bei MDD- und BIP-Patienten signifikant zunahmen, konnte dies bei SCZ-Patienten nicht beobachtet werden. Weiterhin konnte gezeigt werden, dass Patienten, deren depressive Beschwerden nicht relevant zurückgingen, bei Entlassung signifikant höhere NOx- Konzentrationen aufwiesen, was durch die Beobachtung einer signifikant positiven Korrelation zwischen NOx- Serumspiegeln und depressiven Symptomen bei Entlassung unterstützt wurde. Bei der genetischen Untersuchung der Daten fiel auf, dass homozygote Träger des kurzen VNTR-Allels signifikant erhöhte NOx- Konzentrationen besaßen. Diese Ergebnisse blieben bei jenen Trägern auch nach Entlassung signifikant. Insgesamt gibt es Hinweise darauf, dass erhöhte periphere NOx- Metabolitkonzentrationen mit einer Zunahme der Psychopathologie bzw. der Erkrankungsschwere einhergehen könnten, was möglicherweise auf den NOS1 ex1f-VNRT-Polymorphismus zurückzuführen ist. Außerdem zeigten zwei SNPs, welche beide im NOS1AP-Gen lokalisiert sind, bei BIP Patienten signifikant gesteigerte NOx- Werte. Die vorliegenden Ergebnisse deuten darauf hin, dass NO-Signalübertragung und NOS-Genotypen in der Pathogenese psychischer Erkrankungen eine Rolle spielen könnten. Ob diese Veränderungen allerdings kausal mit Krankheitsprozessen zu tun haben oder ob es eher Epiphänomene der Erkrankungen sind, kann mit dieser Studie nicht geklärt werden. Die Genvarianten könnten wiederum bei der Regulierung von peripheren NOx- Konzentrationen von Bedeutung sein. Die Arbeit liefert zudem Hinweise, die Verwendung von NOx als möglichen peripheren Biomarker weiter zu verfeinern und zu untersuchen. Zukünftige Studien, die die Wirksamkeit von NOx- modulierenden Pharmaka untersuchen, könnten davon profitieren, Diagnosegruppen nach Subgruppen einzuteilen, die sowohl NOS Risikogenvarianten als auch periphere NOx- Spiegel im Sinne eines Biomarker beachten.
Das Hepatoblastom ist ein embryonaler Lebertumor, dessen Zellen unterschiedliche unreife Stadien aufweisen. Aufgrund dieser Unreife ist die Identifizierung von Krebsstammzellen erschwert, die in diesem Tumor vermutet und für Rückfälle verantwortlich gemacht werden. In Vorarbeiten konnte eine Krebsstammzellpopulation mit der Kombination der hämatopoetischen Stammzellmarker CD90 und CD34 sowie dem „oval cell“ OV-6-Antikörper in den etablierten Hepatoblastomzelllinien HuH6 und HepG2 detektiert werden. Diese CD34+OV-6+CD90+ Zellen wiesen eine erhöhte Expression von Pluripotenzfaktoren auf und zeichneten sich durch ein erhöhtes Migrationsverhalten aus.
In der hier vorgelegten Arbeit wurden zunächst Tumor-Sphäroid-Assays durchgeführt um diese Population als Krebsstammzellen zu bestätigen, da nur Krebsstammzellen unter den gegebenen Umständen wachsen können. Diese waren im Anschluss wieder in der Lage, in „normalem Medium“ zu differenzieren. Des Weiteren wurden die Zelllinien mit dem Standardtherapeutikum Cisplatin behandelt. Da Krebsstammzellen als chemoresistent gelten, konnte in der überlebenden Zellpopulation auch eine Anreicherung der CD34+OV-6+CD90+ Zellen beobachtet werden. Zusätzlich ließ sich eine weitere CD34+OV-6+CD90+ Population identifizieren, deren Expression aller drei Marker schwächer war und die bei steigenden Cisplatin-Konzentrationen den Großteil der CD34+OV-6+CD90+ Population ausmachte.
Neben den erhöhten Transkriptmengen der Pluripotenzmarker Oct4 und Nanog zeichneten sich die CD34+OV-6+CD90+ Krebsstammzellen durch eine verstärkte Expression der aktivierungsinduzierten Zytidin-Desaminase AID aus. AID wirkt induzierend auf die Transkription beider Pluripotenzmarker. Daher könnte es auch bei Krebsstammzellen eine Rolle bei der Induktion von Pluripotenz und bei ihrer langfristigen Erhaltung spielen. Dies unterstützend legten die verminderten Transkriptmengen der Pluripotenzgene nach einem AID siRNA Knock-Down eine Abhängigkeit des Stammzellcharakters von diesem Faktor nahe. Um einen Effekt von Therapeutika, die inhibitorisch auf AID wirken, auf Krebsstammzellen zu untersuchen, wurden die Zellen mit den DNMT-Inhibitoren Decitabine und Zebularine sowie dem HSP90-Inhibitor Tanespimycin behandelt. Tatsächlich konnte vor allem in den HuH6-Zellen ein verminderter Anteil der Krebsstammzellpopulation durch die drei Substanzen beobachtet werden. Wurden die Zellen im Anschluss mit dem Standardzytostatikum Cisplatin behandelt, führte allerdings eine Vorbehandlung mit Zebularine oder Decitabine zu einer starken Anreicherung von Krebsstammzellen. Dies war vor allem auf Zellen mit dem schwach positiven CD34+OV-6+CD90+ Expressionsprofil zurückzuführen, die chemo-induziert zu sein scheinen. Zwar erscheint die Mehrheit der Tumorzellen eliminiert zu werden, jedoch lassen diese Ergebnisse die Entstehung von chemo-induzierten Krebsstammzellen vermuten, die langfristig für einen Rückfall verantwortlich sein könnten. Daher ist von einer Ergänzung der Therapie mit diesen Substanzen abzusehen. Eine Vorbehandlung mit Tanespimycin hingegen konnte im Vergleich zur alleinigen Cisplatin-Behandlung wirksam den Anteil der Krebsstammzellen reduzieren. Interessanterweise war dies nicht auf einen verstärkten Zelltod der Krebsstammzellen, sondern vielmehr auf eine durch Tanespimycin bewirkte Differenzierung derselben zurückzuführen. Diese spiegelte sich auch in einer verminderten Expression von Pluripotenzmarkern auf mRNA-Ebene im Vergleich zur alleinigen Cisplatin-Behandlung wider.
Somit präsentierte sich Tanespimycin als potenter Inhibitor von Krebsstammzellen. Auch wenn weitere vorklinische und klinische Tests und Untersuchungen erfolgen müssen, stellt Tanespimycin bzw. die Substanzklasse der HSP90-Inhibitoren einen interessanten und vielversprechenden Kandidaten für eine Ergänzung der Standardtherapie vor allem bei behandlungsresistenten und rekurrenten High-Risk-Hepatoblastomen dar.
Bei THS handelt sich um einen operativen Eingriff der Neurochirurgie, bei dem Impulse in tiefere Hirnstrukturen appliziert werden, ohne diese zu beschädigen. Die THS stellt eine etablierte Behandlungsoption bei Bewegungsstörungen von Morbus Parkinson, essentiellem Tremor und Dystonie dar. Zugleich besteht ein zunehmendes Interesse an weiteren Anwendungsmöglichkeiten für neurologische und psychiatrische Erkrankungen. Insoweit wird von einem weiterwachsenden Therapieverfahren in der Neurochirurgie auszugehen sein.
Die bislang implantierten THS-Systeme sind nicht bzw. nur eingeschränkt MRT-fähig.
Aufgrund der in den letzten Jahren zu beobachtenden steigenden Anzahl an Bildgebungen, insbesondere bei MRT-Untersuchungen, stellt sich hier die Frage nach dem Umgang mit einem bildgebenden Verfahren und der Notwendigkeit MRT-fähiger THS-Systeme für das Patientenkollektiv.
Hierzu wurde in der vorliegenden Dissertation analysiert, wie viele der mit THS versorgten Patienten überhaupt ein MRT - nach erfolgreicher Implantation - benötigt haben und zu welchen Konsequenzen dies gegebenenfalls geführt hat.
Die in diesem Zusammenhang retrospektiv gesammelten Patientendaten stammen sowohl aus dem digitalen Patientensystem als auch aus telefonischen Interviews mit Patienten, die seit mindestens 12 Monaten ein THS-System implantiert bekommen haben. Es wurde erfasst, bei wem und aus welchem Grund eine CT- oder MRT-Untersuchung stattgefunden hat. Zusätzlich sind diese Daten von einem unabhängigen Neurologen dahingehend beurteilt worden, ob ein MRT anstatt eines CTs sinnvoller gewesen wäre.
Bei 28 der 54 hier teilnehmenden Patienten ist mindestens eine Bildgebung im Rahmen einer CT- oder MRT-Untersuchung erfolgt. In 16 Fällen ist dabei bei 14 dieser Patienten die Frage aufgekommen, ob ein MRT durchgeführt werden solle. In diesem Zusammenhang sind letztlich sieben MRT-Untersuchungen an sieben Patienten durchgeführt worden, drei kraniale MRT-Untersuchungen und vier Wirbelsäulen MRT-Untersuchungen.
In sieben Fällen bei sechs Patienten hätte der unabhängige Neurologe zu einer MRT-Untersuchung anstatt der durchgeführten CT-Untersuchung geraten.
Von den durchgeführten MRT-Untersuchungen haben alle kranialen sowie zwei Wirbelsäulen MRT-Untersuchungen zu einer konservativen Therapie geführt. Zu einer operativen Therapie haben zwei der durchgeführten Wirbelsäulen MRT-Untersuchungen geführt. Während der in diesem Patientenkollektiv durchgeführten MRT-Untersuchungen ist es zu keiner kritischen Situationen oder Folgeschäden gekommen. Aus Gründen der Patientensicherheit wird trotzdem empfohlen, soweit möglich, bei Patienten mit einem THS-Implantat eine CT-Untersuchung durchzuführen.
Aus den dieser Arbeit zu Grunde liegenden Daten lässt sich konstatieren, dass der Einsatz MRT-fähiger THS-Geräte nicht zwingend geboten ist, dies insbesondere bei jüngeren Patienten jedoch durchaus in Betracht gezogen werden sollte.
Die klinische Pfade zielten in erster Linie darauf ab, die Aufenthaltsdauer zu verkürzen und unnötige Kosten zu sparen, während die Qualität der Pflege erhalten blieb oder verbessert wurde. Bei der laparoskopischen Cholezystektomie gibt es keine ausreichenden Beweise für einen Einfluss auf postoperative Komplikationen.
In dieser retrospektiven Studie wurde die logistische Regression verwendet, um einen Neigungswert zu berechnen, und nach dem Abgleich werden 296 Patienten in beiden Gruppen im Hinblick auf postoperative Komplikationen unter Verwendung des Clavien-Dindo-Klassifizierungssystems als primäres Ziel analysiert. Darüber hinaus wurden sekundäre Ziele wie Aufenthaltsdauer, Einhaltung und Abweichung vom klinischen Pfad in Bezug auf die Entlassung von Patienten analysiert. Das relative Risiko des primären Ergebnisses wurde berechnet und mit dem E-Wert als Ansatz für Sensitivitätstests verglichen.
Aufgrund des obligatorischen Teil der klinischen Pfad bei den Patienten betrug die Compliance 100 Prozent. In 16% der Fälle trat eine Abweichung vom Pfad in Bezug auf die geplante Entlassung des Patienten am zweiten Tag nach der Operation auf. Nach Anpassung um potenzielle Faktoren beträgt das relative Risiko beim Vergleich der Clavien-Dindo-Komplikationsbewertung 0 versus 1-4 ist 1,64 (95% CI 0,87; 3,11), was nicht signifikant unterschiedlich ist (p = 0,127).
Nach Matching beträgt die Verweildauer 3,69 Tage ohne bzw. 3,26 Tage mit dem klinischen Pfad.
Vergleich zu bereits implementierter strukturierter Standardoperationsverfahren kann ein klinischer Pfad postoperative Komplikationen nicht reduzieren.
Dennoch betrachten wir unseren klinischen Pfad als ein äußerst wertvolles Instrument für die interdisziplinäre Verwaltung des Krankenhausaufenthalts des Patienten unter der Aufsicht eines erfahrenen Chirurgen.
Dieser Arbeit lag die Fragestellung zugrunde, welchen Einfluss die jeweilige Transfusionsstrategie auf AML-Patienten unter intensiver Induktionschemotherapie hat.
Dafür wurde ein am Universitätsklinikum Frankfurt zwischen 2007 und 2018 behandeltes Kollektiv von 352 Patienten untersucht. So erfolgte hier bis ins Jahr 2015 hinein die Transfusion eines EK ab einem Hb-Wert <8g/dl und nach Änderung des Transfusionstriggers ab einem Hb <7g/dl. AML-Patienten aus dem Jahr 2015 – dem Jahr der Änderung der Transfusionsregel – wurden von weiterer Untersuchung ausgeschlossen, um zwei klar abgrenzbare Kohorten erhalten zu können.
Es zeigte sich, dass die weniger restriktive Transfusionskohorte unter Induktionschemotherapie einen durchschnittlich um 1g/dl höheren Hb-Wert aufwies und früher als die restriktive Kohorte transfundiert wurde. Die Anzahl an Fiebertagen, CRP-Werte, Aufenthalte auf Intensivstation sowie die Dauer des Krankenhausaufenthaltes betrachtend, zeigte sich hingegen kein signifikanter Unterscheid zwischen beiden Kohorten.
Basierend auf unserer Arbeit ergeben sich keine Hinweise dafür, dass die restriktive Transfusionspraxis einer weniger restriktiven für AML-Patienten unterlegen ist. Limitierend auf die Aussagekraft der Ergebnisse wirken sich dabei die retrospektive Natur der Arbeit sowie die zeitliche Verschiebung der Behandlungszeiträume beider Kohorten aus.
Ergebnisse der bislang ausstehenden randomisierten Studien, die den Einfluss unterschiedlicher Transfusionsregimes auf Patienten mit hämatologischen Krankheiten untersuchen, sind in Bälde zu erwarten. Die bereits vorliegenden Pilotstudien und Ergebnisse der TRIST-Studie decken sich mit der von uns beobachteten Nicht-Unterlegenheit der restriktiven Transfusionspraxis für ein hämatoonkologisches Patientenkollektiv, sodass es abzuwarten gilt, ob sich dies auch in weiteren größeren randomisierten und kontrollierten Studien beweisen kann.
Das Harnblasenkarzinom ist einer der häufigsten Tumoren weltweit. Insbesondere die muskelinvasiven Tumoren haben eine schlechte Prognose und stellen sich morphologisch sehr unterschiedlich dar. Diese Heterogenität wird bislang bei Therapieentscheidungen nicht beachtet. Um Patienten zukünftig individuell auf den vorliegenden Subtyp des muskelinvasiven Harnblasenkarzinoms (MIBC, muscle invasive bladder cancer) behandeln zu können und dadurch unnötige Belastungen durch Chemotherapien vermeiden zu können, ist eine einfache und kostengünstige Diagnostik erforderlich. Das Ziel der vorliegenden Arbeit war es, den Einsatz bestimmter immunhistochemischer Färbungen als ein mögliches diagnostisches Routineverfahren zur Bestimmung der vorliegenden Subtypen auszutesten. Hierzu wurde die Expression von „luminalen“ und „basalen“ Proteinen mit histologischen Subtypen des MIBCs korreliert. In einem zweiten Schritt wurde der Einfluss auf das Überleben mit und ohne adjuvante Chemotherapie untersucht.
Es wurden insgesamt 181 Tissue-Microarray-Spots analysiert. Alle histologischen Patientenproben sowie klinischen Daten stammten aus dem Universitätsklinikum Frankfurt am Main. Aus den entsprechenden Gewebeblöcken wurden Stanzen zur Erstellung eines Tissue-Microarrays (TMAs) entnommen. Diese wurden konventionell angeschnitten und histologisch mit Hämatoxylin-Eosin (H/E) sowie immunhistochemisch mit Cytokeratin 5/6 (CK5/6), Cytokeratin 20 (CK20), Glutamyl Aminotransferase-Untereinheit A bindendem Protein 3 (GATA3), Tumorsuppressor- protein p53 und Synaptophysin (SYNAPT) gefärbt.
Anhand der H/E-Schnitte wurden die vorliegenden histologischen Subtypen lichtmikroskopisch bestimmt und es folgte eine statistische Auswertung der Färbeergebnisse. Die deskriptive statistische Analyse zeigte insbesondere für die beiden Färbungen CK5/6 und GATA3 signifikante Ergebnisse. Deshalb wurden in einem zweiten Schritt alle Fälle der Studienkohorte den bekannten vier Gruppen: CK5/6 positiv, GATA3 positiv, doppelt positiv und doppelt negativ zugeordnet und näher untersucht. Es folgte eine Überlebensanalyse nach Kaplan Meier Schätzer sowie uni- und multivariate Analysen.
Die Ergebnisse zeigten eine Assoziation der Expression von CK5/6 mit einer squamösen Differenzierung (96%) und eine Assoziation der Expression von GATA3 mit einer mikropapillären Differenzierung (100%). Die adjuvante Chemotherapie ging mit einem Überlebensvorteil (HR 0,15 95%KI 0,1-0,3; p<0,001) der Patienten mit MIBC einher. Immunhistochemisch doppelt negative Patienten mit MIBC wiesen ein verringertes Gesamtüberleben auf (HR 4,96; 95%KI 1,6-15,6; p=0,006); in der Gruppe der doppelt negativen MIBCs fanden sich fünf verschiedene histologische Subtypen. Zusammenfassend lässt sich sagen, dass die immunhistochemische Klassifizierung des MIBCs mit histologischen Subtypen assoziiert ist und dabei helfen kann, Fälle in der pathologischen Routine in „luminal“ und „basal“ zu unterteilen. Jedoch ist ein auf zwei Markern basierendes Klassifizierungssystem nicht ausreichend, um die Heterogenität des MIBCs abzubilden und die Basis für Therapieentscheidungen zu bilden.
Polyunsaturated fatty acids (PUFAs) play essential roles in mediating inflammation and its resolution. PUFA metabolites generated by the cytochrome P450 (CYP) - soluble epoxide hydrolase (sEH) axis are known to regulate macrophage activation/polarization but little is known about their role in the resolution of inflammation. Monocytes were isolated from murine bone marrow or human peripheral blood and differentiated to naïve macrophages (M0). Thereafter cells were polarized using LPS and IFNγ (M1), IL-4 (M2a), or TGFβ1 (M2c). Gene expression was analyzed by RNA sequencing, RT-qPCR and Western blotting. Phagocytosis of zymosan and oxo-LDL were also assessed in vitro. Zymosan-induced peritonitis combined with immune cell profiling was used to evaluate the resolution of inflammation in vivo. The expression of sEH was comparable in M0, M1 and M2a macrophages but markedly elevated in M2c polarized cells. The increase in sEH expression elicited by TGFβ relied on the TGFβ receptor ALK5 and the phosphorylation of SMAD2, which was able to bind to the sEH promoter. In macrophages lacking sEH, M2c polarization was incomplete and characterized by lower levels of pro-resolving phagocytosis associated receptors (Tlr2 and Mrc1), as well as higher levels of the pro-inflammatory markers; Nlrp3, IL-1β and TNFα. Fitting with the failure to upregulate phagocytosis associated receptors, the uptake of zymosan and ox-LDL was less efficient in M2c macrophages from sEH-/- mice. The latter animals also demonstrated a retarded resolution of inflammation (zymosan-induced peritonitis) in vivo with fewer resident macrophages and recruited macrophages. PUFA profile analysis indicated decreased sEH substrates e.g., 11, 12-EET, as well as increased sEH products e.g., 11, 12-DHET, indicating an increased sEH activity in M2c macrophages. Taken together, our data indicates that sEH expression is required for the effective M2c polarization of macrophages and thus the resolution of inflammation.
Test-Retest-Reliabilität der Präpulsinhibition (PPI) und PPI-Korrelation mit dem Arbeitsgedächtnis
(2023)
Sensomotorisches Gating – ein Mechanismus zur Filterung des sensorischen Inputs und zur Regulierung des motorischen Outputs – wird experimentell durch die Präpulsinhibition (PPI) der akustisch ausgelösten Schreckreaktion (ASR) operatio-nalisiert. Frühere Studien deuten auf eine hohe Test-Retest-Reliabilität der PPI und eine mögliche Korrelation mit dem Arbeitsgedächtnis (engl. Working Memory (WM)) hin. Ziel dieser Studie war es, die Test-Retest-Reliabilität der PPI bei ge-sunden Menschen und ihre Korrelation mit der Leistung des WM zu überprüfen. Hier wurde ein akustisches Schreckreiz-PPI-Paradigma mit vier verschiedenen Präpuls-Intensitäten (64, 68, 72 und 76 dB(A)) und zwei verschiedene WM-Aufgaben (n-back, Change-Detection-Task (CDT)) verwendet. Es konnte eine ho-he Retest-Reliabilität der PPI mit einer mittleren Intraklassenkorrelation (engl. In-traclass Correlation (ICC)) von >.80 und eine signifikante positive Korrelation der PPI mit der n-back-, aber nicht mit der CDT-Leistung bestätigt werden. Eine detail-lierte Analyse zeigte, dass die PPI über alle Präpulsintensitäten hinweg sowohl mit den 2-back- als auch mit den 0-back-Bedingungen signifikant korrelierte, was auf eine Regulation durch konditionsübergreifende Prozesse (z. B. Aufmerksamkeit) schließen lässt. Wird jedoch die 0-back-Komponente aus den 2-back-Daten aus-partialisiert, sind spezifische und signifikante Korrelationen mit der Arbeitsgedächt-nisleistung für die 76 dB(A) PPI-Bedingung zu finden. Mit der vorliegenden Studie konnte die hohe Test-Retest-Reliabilität der PPI beim Menschen bestätigt und die Korrelation mit der Arbeitsgedächtnisleistung validiert und erweitert werden.