No / Inflammatory changes, characterized by an increase in pro-inflammatory cytokine production and up-regulation of the corresponding signaling pathways, have been described in the brains of aged rats and rats treated with the potent immune modulatory molecule lipopolysaccharide (LPS). These changes have been coupled with a deficit in long-term potentiation (LTP) in hippocampus. The evidence suggests that anti-inflammatory agents, which attenuate the LPS-induced and age-associated increase in hippocampal interleukin-1ß (IL-1ß) concentration, lead to restoration of LTP. Here we report that atorvastatin, a member of the family of agents that act as inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase, exerts powerful anti-inflammatory effects in brain and that these effects are mediated by IL-4 and independent of its cholesterol-lowering actions. Treatment of rats with atorvastatin increased IL-4 concentration in hippocampal tissue prepared from LPS-treated and aged rats and abrogated the age-related and LPS-induced increases in pro-inflammatory cytokines, interferon-¿ (IFN¿) and IL-1ß, and the accompanying deficit in LTP. The effect of atorvastatin on the LPS-induced increases in IFN¿ and IL-1ß was absent in tissue prepared from IL-4¿/¿ mice. The increase in IL-1ß in LPS-treated and aged rats is associated with increased microglial activation, assessed by analysis of major histocompatibility complex II expression, and the evidence suggests that IFN¿ may trigger this activation. We propose that the primary effect of atorvastatin is to increase IL-4, which antagonizes the effects of IFN¿, the associated increase in microglial activation, and the subsequent cascade of events.

Red seaweeds are reported to represent the largest group of algae, with more species accounted for than the combination of brown and green seaweeds. Due to the high amount of polysaccharides in red seaweeds, they are mainly utilized for commercial agar and carrageenan production in industry. However, increasing studies indicate other valuable compounds such as lipids and polyphenols could be potential utilized for multiple human needs (e.g., drug development) (1, 2). With increasing studies demonstrating the potential health benefits of seaweed components, two red seaweed species commonly consumed in Asia, hong qı´ lı´n c a`i (HQL), Eucheuma sp and zhe` gu¯ ca`i (ZGC), Caloglossa leprieurii, were investigated on to determine the anti-inflammatory effects of their extractable lipophilic bioactives (ELB) and bound lipophilic bioactives (BLB) in lipopolysaccharide( LPS)-treated RAW 264.7 macrophages. The chemical composition of ELB and BLB was characterized in terms of total phenolic content (TPC), total flavonoid content (TFC), total tannin content (TTC), oxygen radical absorbance capacity (ORAC), and etc. Six phenolic compounds were identified in ZGC extracts and one was detected in HQL. All extracts inhibited the nitric oxide (NO) production in LPS-induced macrophages, which was associated with downregulation of iNOS and COX-2 protein expression and up-regulation of HQ-1 and NQO1 protein expression. Overall, our results showed that both ELB and BLB in HQL and ZGC seaweeds presented potential anti-inflammatory activities. These results warrant future investigations to determine the mode of actions of red seaweed bioactives and their efficacy in humans.

Lactobacillus delbrueckii est une bactérie laitière non commensale qui transite dans notre tractus gastro-intestinal du fait de son utilisation dans les produits laitiers fermentés, notamment les yaourts. Nous avons cherché à déterminer les profils immuno-modulateurs de 59 souches de L. delbrueckii subsp. lactis et subsp. bulgaricus. Nous avons d'abord criblé notre panel de souches sur leur capacités à inhiber l'activation de NF-kB par le TNF-α dans des cellules HT-29. Nous avons ainsi pu mettre en évidence une variabilité des capacités immuno-modulatrices chez ces souches en repérant des souches aux capacités anti- ou pro-inflammatoires. Nous avons retenu les deux souches les plus anti-inflammatoires (L. delbrueckii subsp. lactis CNRZ327 et 333) et avons observé qu'elles perdaient leurs effets après traitement à la trypsine suggérant que les effecteurs inhibant l'activation de NF-kB soient de nature protéique. Nous avons pu aussi déterminer que cette inhibition passait par la réduction de la phosphorylation d'IkB affectant la translocation de NF-kB dans le noyau. Nous avons ensuite testé ces deux souches dans un modèle murin de colite induite au DSS. La souche CNRZ327 a amélioré de façon significative les dommages induits par le DSS confirmant ainsi ses effets anti-inflammatoires in vivo. Nous avons ensuite tenté de déterminer les mécanismes des effets anti-inflammatoires de la CNRZ327. Cette souche i) augmente les taux d'IgA sécrétoires; ii) module la production de TGF-β dans le colon et de l'IL-6 et de l'IL-10 dans la rate; iii) augmente les cellules T régultrices CD4+Foxp3+ dans la rate et les ganglions cécaux et iv) module la fréquence des cellules dendritiques TLR2+2 and TLR4+. Nos travaux constituent la première démonstration d'effets immuno-modulateurs d'une souche de lactobacille laitier. Les lactobacilles laitiers très présents dans nos aliments laitiers fermentés peuvent ainsi moduler notre réponse immune et influencer ainsi notre santé. / The work presented in this thesis aims to review the present knowledge on bacteria-host interactions in the GI tract, and to demonstrate the immune modulatory effects of Lactobacillus delbrueckii, a non-commensal dairy bacterium that is in constant transit in our GI tract through the ingestion of fermented food products, like milk and cheese.The main results presented in this thesis can be divided into two parts. In the first part we revisited the anti-inflammatory properties of L. delbrueckii by screening a collection of different strains for their ability to inhibit TNF-α-induced NF-kB activation in an IEC cell line. Our results demonstrated the existence of inter strain variation for immune modulation properties in the L. delbrueckii species. Two of the most effective strains completely lost their ability to suppress NF-kB after trypsin treatment, indicating that the bacterial effectors involved in the NF-kB modulation are proteinaceous in nature. We also showed that L. delbrueckii inhibits NF-kB activation by reducing the phosphorylation of IkB, which would affect translocation of NF-kB to the nucleus. Based on our in vitro results, we selected three strains for tests in a DSS model of experimental colitis. One of the strains tested, L. delbrueckii subsp.Lactis CNRZ327 (Lb CNRZ327) consistently improved the DSS-induced damage, thus confirming its anti-inflammatory properties in vivo. The second part of my thesis was dedicated to the study of the mechanisms involved in the anti-inflammatory effects of L. delbrueckii in the DSS-model of colitis. Lb CNRZ327 showed a tendency to increases s-IgA levels, modulates the production of TGF-β in colonic tissue, and of IL-6 and IL-10 in the spleen, expands the frequency of CD4+Foxp3+ regulatory T cells in the spleen and CLN during colitis and modulates the frequencies of TLR2 and TLR4 expressing dendritic cells in the cecal lymph nodes. TLR2 appears to be involved in Lb CNRZ327 recognition and affect IL-12 production in BMDM. This is the first demonstration of systemic immune modulation effects exerted by a dairy Lactobacillus. The results of this study show that dairy lactobacilli that often are part of a regular diet can modulate innate immune responses and may thus affect health more than generally thought.

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Alžbeta Horvátová Supervisor: prof. PharmDr. Petr Pávek PhD. Title of diploma thesis: Anti-inflammatory effects of ursodeoxycholyl lysophosphatidylethanolamide on THP-1 human macrophages via Toll-like receptor 4 Nonalcoholic steatohepatitis (NASH) became the most common liver disease in developed countries. It is well-known that the level of protectant phosphatidylcholine (PC) is decreased in NASH. The bile acid-phospholipid conjugate ursodeoxycholyl lysophosphatidylethanolamide (UDCA-LPE) was designed in order to specifically deliver PC to hepatocytes. However, previous studies have proved that UDCA-LPE possesses its proper hepatoprotectant capacity and exhibits anti-apoptotic, anti-inflammatory, anti-fibrotic properties and also improved steatosis and hyperlipidaemia in various models in vivo. These effects may be mediated secondary through modulation of immune system. Therefore, in order to dissect if UDCA-LPE directly influences immune cells in vitro, release of pro-inflammatory cytokines TNFα, IL-6 and IL-1β in LPS-induced THP-1-derived human macrophages was measured by ELISA. Moreover, effects of UDCA-LPE on MAPK signalling pathways and nuclear translocation of NFκB were...

A cirurgia endoscópica transluminal por orifícios naturais (NOTES) é uma nova modalidade cirúrgica caracterizada pela ausência de incisões cutâneas. Diferentes vias de acesso para a NOTES já foram testadas, porém, existem poucos estudos sobre os efeitos imunomoduladores da via transretal e de sua associação com as células-tronco mesenquimais (MSCs) como adjuvante nos efeitos anti-inflamatórios e na cicatrização retal. Este trabalho objetivou avaliar a exequibilidade da técnica NOTES transretal, os efeitos imunomodulatórios e regenerativos desta abordagem cirúrgica e sua associação com as MSCs em um modelo experimental suíno. A tese foi divida em quatro capítulos sendo que o primeiro trata de uma revisão bibliográfica sobre as vias de acesso para a NOTES e suas implicações. O segundo objetivou testar a exequibilidade da via transretal em três suínos, o que demonstrou ser um acesso alternativo para procedimentos abdominais. O terceiro capítulo avalia a resposta inflamatória da técnica NOTES e da sua associação com MSC derivadas de tecido adiposo. Para isso, 11 suínos foram submetidos a NOTES transretal e distribuídos aleatoriamente em dois grupos: GNOTES e GNOTES+MSC. Amostras de sangue foram coletadas em diferentes tempos para exames de hemograma, albumina, proteína C reativa (PCR) e interleucina- 6 (IL-6). Não houveram alterações entre os grupos quanto aos valores de hemograma, albumina e PCR, sendo que a IL-6 foi indetectável em todos os animais. Histologicamente, foi observado menor quantidade de células inflamatórias no GNOTES+MSC na camada muscular. Estes dados sugerem que tanto a técnica NOTES quanto as MSCs atenuam os efeitos inflamatórios do procedimento. Um prolapso retal ocorrido como complicação da técnica NOTES transretal foi detalhadamente descrito no quarto capítulo. Assim, a técnica NOTES associada ou não às MSCs no sítio da incisão apresenta mínima resposta inflamatória em suínos submetidos à peritoneoscopia transretal. / Natural orifice transluminal endoscopic surgery (NOTES) is an evolving surgical approach to be performed without skin incisions. Different access sites have been tested, however, studies regarding immunomodulation of the transrectal approach and its association with mesenchymal stem cells (MSC) are still lacking. The viability of transrectal NOTES and the associated effects of MSC local therapy on rectal healing and inflammation were tested in a porcine model. The results of the study are depicted in fours chapters. The first chapter presents a literature review and state-of-the-art on the accesses for NOTES. The subsequent paper describes the transrectal approach performed in three pigs and its viability as an alternative access to abdominal endoscopic procedures. In the third chapter, the evaluation of the inflammatory response with transrectal NOTES and adipose tissue MSC therapy is presented. For that, 11 pigs were either submitted to NOTES only or NOTES with local MSC application. Blood samples, peritoneal fluid, and post-mortem rectal tissue specimens were collected. Total blood count, albumin and C-reactive protein showed no significant differences between groups. Interleukin-6 was undetectable in both groups. Histological evaluations demonstrated less inflammatory cells in the muscular layer of rectum of the animals where NOTES was associated with MSC. A possible lessening of the immune response may be seen with NOTES with or without MSCs; however, the presence of MSC attenuate the local inflammatory reaction in the muscular layer of porcine rectum after transrectal NOTES. A rectal prolapse that occurred after transrectal NOTES in one pig is reported in the last chapter. The transrectal NOTES technique with or without the MSCs at the incision site shows minimal inflammatory response in porcine.

A cirurgia endoscópica transluminal por orifícios naturais (NOTES) é uma nova modalidade cirúrgica caracterizada pela ausência de incisões cutâneas. Diferentes vias de acesso para a NOTES já foram testadas, porém, existem poucos estudos sobre os efeitos imunomoduladores da via transretal e de sua associação com as células-tronco mesenquimais (MSCs) como adjuvante nos efeitos anti-inflamatórios e na cicatrização retal. Este trabalho objetivou avaliar a exequibilidade da técnica NOTES transretal, os efeitos imunomodulatórios e regenerativos desta abordagem cirúrgica e sua associação com as MSCs em um modelo experimental suíno. A tese foi divida em quatro capítulos sendo que o primeiro trata de uma revisão bibliográfica sobre as vias de acesso para a NOTES e suas implicações. O segundo objetivou testar a exequibilidade da via transretal em três suínos, o que demonstrou ser um acesso alternativo para procedimentos abdominais. O terceiro capítulo avalia a resposta inflamatória da técnica NOTES e da sua associação com MSC derivadas de tecido adiposo. Para isso, 11 suínos foram submetidos a NOTES transretal e distribuídos aleatoriamente em dois grupos: GNOTES e GNOTES+MSC. Amostras de sangue foram coletadas em diferentes tempos para exames de hemograma, albumina, proteína C reativa (PCR) e interleucina- 6 (IL-6). Não houveram alterações entre os grupos quanto aos valores de hemograma, albumina e PCR, sendo que a IL-6 foi indetectável em todos os animais. Histologicamente, foi observado menor quantidade de células inflamatórias no GNOTES+MSC na camada muscular. Estes dados sugerem que tanto a técnica NOTES quanto as MSCs atenuam os efeitos inflamatórios do procedimento. Um prolapso retal ocorrido como complicação da técnica NOTES transretal foi detalhadamente descrito no quarto capítulo. Assim, a técnica NOTES associada ou não às MSCs no sítio da incisão apresenta mínima resposta inflamatória em suínos submetidos à peritoneoscopia transretal. / Natural orifice transluminal endoscopic surgery (NOTES) is an evolving surgical approach to be performed without skin incisions. Different access sites have been tested, however, studies regarding immunomodulation of the transrectal approach and its association with mesenchymal stem cells (MSC) are still lacking. The viability of transrectal NOTES and the associated effects of MSC local therapy on rectal healing and inflammation were tested in a porcine model. The results of the study are depicted in fours chapters. The first chapter presents a literature review and state-of-the-art on the accesses for NOTES. The subsequent paper describes the transrectal approach performed in three pigs and its viability as an alternative access to abdominal endoscopic procedures. In the third chapter, the evaluation of the inflammatory response with transrectal NOTES and adipose tissue MSC therapy is presented. For that, 11 pigs were either submitted to NOTES only or NOTES with local MSC application. Blood samples, peritoneal fluid, and post-mortem rectal tissue specimens were collected. Total blood count, albumin and C-reactive protein showed no significant differences between groups. Interleukin-6 was undetectable in both groups. Histological evaluations demonstrated less inflammatory cells in the muscular layer of rectum of the animals where NOTES was associated with MSC. A possible lessening of the immune response may be seen with NOTES with or without MSCs; however, the presence of MSC attenuate the local inflammatory reaction in the muscular layer of porcine rectum after transrectal NOTES. A rectal prolapse that occurred after transrectal NOTES in one pig is reported in the last chapter. The transrectal NOTES technique with or without the MSCs at the incision site shows minimal inflammatory response in porcine.

A cirurgia endoscópica transluminal por orifícios naturais (NOTES) é uma nova modalidade cirúrgica caracterizada pela ausência de incisões cutâneas. Diferentes vias de acesso para a NOTES já foram testadas, porém, existem poucos estudos sobre os efeitos imunomoduladores da via transretal e de sua associação com as células-tronco mesenquimais (MSCs) como adjuvante nos efeitos anti-inflamatórios e na cicatrização retal. Este trabalho objetivou avaliar a exequibilidade da técnica NOTES transretal, os efeitos imunomodulatórios e regenerativos desta abordagem cirúrgica e sua associação com as MSCs em um modelo experimental suíno. A tese foi divida em quatro capítulos sendo que o primeiro trata de uma revisão bibliográfica sobre as vias de acesso para a NOTES e suas implicações. O segundo objetivou testar a exequibilidade da via transretal em três suínos, o que demonstrou ser um acesso alternativo para procedimentos abdominais. O terceiro capítulo avalia a resposta inflamatória da técnica NOTES e da sua associação com MSC derivadas de tecido adiposo. Para isso, 11 suínos foram submetidos a NOTES transretal e distribuídos aleatoriamente em dois grupos: GNOTES e GNOTES+MSC. Amostras de sangue foram coletadas em diferentes tempos para exames de hemograma, albumina, proteína C reativa (PCR) e interleucina- 6 (IL-6). Não houveram alterações entre os grupos quanto aos valores de hemograma, albumina e PCR, sendo que a IL-6 foi indetectável em todos os animais. Histologicamente, foi observado menor quantidade de células inflamatórias no GNOTES+MSC na camada muscular. Estes dados sugerem que tanto a técnica NOTES quanto as MSCs atenuam os efeitos inflamatórios do procedimento. Um prolapso retal ocorrido como complicação da técnica NOTES transretal foi detalhadamente descrito no quarto capítulo. Assim, a técnica NOTES associada ou não às MSCs no sítio da incisão apresenta mínima resposta inflamatória em suínos submetidos à peritoneoscopia transretal. / Natural orifice transluminal endoscopic surgery (NOTES) is an evolving surgical approach to be performed without skin incisions. Different access sites have been tested, however, studies regarding immunomodulation of the transrectal approach and its association with mesenchymal stem cells (MSC) are still lacking. The viability of transrectal NOTES and the associated effects of MSC local therapy on rectal healing and inflammation were tested in a porcine model. The results of the study are depicted in fours chapters. The first chapter presents a literature review and state-of-the-art on the accesses for NOTES. The subsequent paper describes the transrectal approach performed in three pigs and its viability as an alternative access to abdominal endoscopic procedures. In the third chapter, the evaluation of the inflammatory response with transrectal NOTES and adipose tissue MSC therapy is presented. For that, 11 pigs were either submitted to NOTES only or NOTES with local MSC application. Blood samples, peritoneal fluid, and post-mortem rectal tissue specimens were collected. Total blood count, albumin and C-reactive protein showed no significant differences between groups. Interleukin-6 was undetectable in both groups. Histological evaluations demonstrated less inflammatory cells in the muscular layer of rectum of the animals where NOTES was associated with MSC. A possible lessening of the immune response may be seen with NOTES with or without MSCs; however, the presence of MSC attenuate the local inflammatory reaction in the muscular layer of porcine rectum after transrectal NOTES. A rectal prolapse that occurred after transrectal NOTES in one pig is reported in the last chapter. The transrectal NOTES technique with or without the MSCs at the incision site shows minimal inflammatory response in porcine.