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Phagocytosis is an inherent function of tissue macrophages for the removal of apoptotic cells and cellular debris during acute and chronic injury; however, the dynamics of this event during fibrosis development is unknown. We aim to prove that during the development of kidney fibrosis in the unilateral ureteral obstruction (UUO) model, there are some populations of macrophage with a reduced ability to phagocytose, and whether the infusion of a population of phagocytic macro-phages could reduce fibrosis in the murine model UUO. For this purpose, we have identified the macrophage populations during the development of fibrosis and have characterized their phagocytic ability and their expression of CPT1a. Furthermore, we have evaluated the therapeutic effect of macrophages overexpressing CPT1a with high phagocytic skills. We evidenced that the macro-phage population which exhibits high phagocytic ability (F4/80low-CD11b) in fibrotic animals de-creases during the progression of fibrosis while the macrophage population with lower phagocytic ability (F4/80high-CD11b) in fibrotic conditions, conversely, increases and CPT1a macrophage cell therapy with a strengthening phagocytic ability is associated with a therapeutic effect on kidney fibrosis. We have developed a therapeutic approach to reduce fibrosis in the UUO model by enrichment of the kidney resident macrophage population with a higher proportion of exogenous phagocytic macrophages overexpressing CPT1a., This research was funded by grants from the Ministerio de Economía y Competitividad, reference SAF2015–67770-R, awarded to G.H.; from the Fondo de Investigación Sanitaria (FIS), reference PI20/00900, awarded to G.H and the program Retos Colaboracion, reference: RTC2019- 007197-1, awarded G.H. more...

We propose the use of a peripheral blood mononuclear cell therapy based on cell NGAL release to be used in the clinical setting for acute kidney injury (AKI) and the derived fibrosis. First, we designed a procedure whereby PBMC overexpress NGAL and anti-inflammatory agents when subjected to repetitive anoxia/reoxygenation (PBMC (A/R)). Using an in vivo AKI model, we observed that PBMC(A/R) reduces BUN and creatinine levels in blood and inflammation, enhances anti-inflammation, induces proliferation of tubular epithelial cells and reduces AKI-induced fibrosis. Flow cytometry analysis evidenced that monocytes are the only cells accumulated in the injured kidney and phenotype analysis of freshly isolated kidney macrophages, revealed that the healing phenotype is maintained the time needed for recovery. NGAL release from PBMC(A/R) determines the beneficial effect of the therapy since administration of a NGAL antibody previous to the therapy or injection of PBMC(A/R) obtained from NGAL KO animals abolished the beneficial effects. CD11b–NGAL positive cells were enhanced in tissue after PBMC (A/R) therapy and were produced by the injected monocytes. In an in vitro model with tubular epithelial cells (NRK52e) we proved that NGAL release by PBMC(A/R) induced epithelial proliferation and activation of PI3K/Akt pathway., This research was funded by grants RTC2019-0079197-1 funded by Ministerio de Ciencia e Innovacion/Agencia ´ Estatal de Investigacion ´ MCIN/AEI/10.13039/501100011033, awarded to G.H and grant PI20/ 00900 funded by Fondo de Investigacion ´ Sanitaria (FIS), awarded to G. H. more...

© 2020 by the authors., Diabetic nephropathy (DN) is the leading cause of end-stage renal disease globally. The primary initiating mechanism in DN is hyperglycemia-induced vascular dysfunction, but its progression is due to different pathological mechanisms, including oxidative stress, inflammatory cells infiltration, inflammation and fibrosis. Macrophages (Mφ) accumulation in kidneys correlates strongly with serum creatinine, interstitial myofibroblast accumulation and interstitial fibrosis scores. However, whether or not Mφ polarization is involved in the progression of DN has not been adequately defined. The prevalence of the different phenotypes during the course of DN, the existence of hybrid phenotypes and the plasticity of these cells depending of the environment have led to inconclusive results. In the same sense the role of the different macrophage phenotype in fibrosis associated or not to DN warrants additional investigation into Mφ polarization and its role in fibrosis. Due to the association between fibrosis and the progressive decline of renal function in DN, and the role of the different phenotypes of Mφ in fibrosis, in this review we examine the role of macrophage phenotype control in DN and highlight the potential factors contributing to phenotype change and injury or repair in DN, This study was supported by grants from Ministerio de Economiía y Competitividad, reference SAF2015–67770-R, awarded to G.H. more...

Background: Dyslipidemia causes renal damage; however, the detailed molecular mechanism has not been clarified. It is known that carnitine palmitoyl transferase 1-a (CPT1a) gene encodes an enzyme involved in fatty acid oxidation and, therefore, lipid content. In the present study, we investigated whether the accumulation of lipids induced by 7-ketocholesterol (7-KC) in tubular epithelial cells produce a fibrotic and inflammatory response through CPT1a. Methods: Using an epithelial cell line, NRK-52E, we determine if intracellular accumulation of 7-KC modulates profibrotic and inflammatory events through CPT1a gene expression. In addition, the direct effects of CPT1a genetic modification has been studied. Results: Our results revealed that high levels of 7-KC induce increased expression of CPT1a, TGF-β1, α- SMA and NLRP3 that was correlated with lipid content. GW3965 treatment, which have shown to facilitate the efflux of cholesterol, eliminated the intracellular lipid droplets of 7-KC laden cells and decreased the expression of CPT1a, TGF-β1, α- SMA and NLRP3. Furthermore, CPT1a Knockdown and C75 pre-treatment increased lipid content but decreased TGF-β1, α- SMA and NLRP3. Conclusions: Our findings reveal that the profibrotic effect of 7-KC on renal epithelial cells are mediated by CPT1a overexpression, which acts on TGF-β1, α-SMA and NLRP3. Thus, CPT1a downregulation protects against 7-KC-induced fibrosis in tubular epithelial cells by downregulating TGF-β1, α- SMA and NLRP3., This study was supported by grants from Ministerio de Economía y Competitividad, awarded to G.H reference SAF2015-67770-R.P.C is performing the studies under the PhD program in Medicine ofthe Universitat de Barcelona, funded by the mentioned grant more...

[Background] Macrophage are specialized cells that contributes to the removal of detrimental contents via phagocytosis. Lipid accumulation in macrophages, whether from phagocytosis of dying cells or from circulating oxidized low-density lipoproteins, alters macrophage biology and functionality. It is known that carnitine palmitoyl transferase 1-a (CPT1a) gene encodes an enzyme involved in fatty acid oxidation and, therefore, lipid content. However, the potential of CPT1a to activate macrophage phagocytic function have not been elucidated., [Methods] Using a murine macrophage cell line, RAW264.7, we determine if intracellular accumulation of 7-ketocholesterol (7-KC) modulates macrophage phagocytic function through CPT1a gene expression. In addition, the effects of CPT1a genetic modification on macrophage phenotype and phagocytosis has been studied., [Results] Our results revealed that CPT1a gene expression decreased by the accumulation of 7-KC at the higher dose of 7-KC. This was concomitant with an impair ability to phagocytize bioparticles and an inflammatory phenotype. GW3965 treatment, which have shown to facilitate the efflux of cholesterol, eliminated the intracellular lipid droplets of 7-KC-laden macrophages, increased the gene expression of CPT1a, diminished the gene expression of the inflammatory marker iNOS and restored macrophage phagocytosis. Furthermore, CPT1a Knockdown per se was detrimental for macrophage phagocytosis whereas transcriptional activation of CPT1a heightened the uptake of bioparticles., [Conclusions] Altogether, our findings indicate that downregulation of CPT1a by lipid content modulates macrophage phagocytosis and inflammatory phenotype., This study was supported by grants from Ministerio de Economía y Competitividad, reference SAF2015–67770-R, awarded to G.H and from Fondo de Investigación Sanitaria (FIS), reference PI17/01411, awarded to A.S. more...

To evaluate the diagnostic utility of routine gastric biopsies taken at random versus targeted biopsies with methylene blue staining for the diagnosis of intestinal metaplasia.We performed a validation study in 376 patients. We performed 2 antral biopsies, 2 gastric body biopsies, 1 biopsy for urease test and additional biopsies if demanded. One hundred and one patients underwent 2 biopsies after methylene blue staining. A total of 1,486 biopsies were analyzed. Frequencies of histological and endoscopic diagnosis and the validation of endoscopic diagnosis of intestinal metaplasia with and without staining against histology, which followed updated Sydney and Vienna criteria, were determinedMetaplasia was found in 37.23% ofcases and in 6.38% was incomplete in antrum and body, moderate or severe. The endoscopic diagnosis had: sensitivity 46%, specificity 91%, positive predictive value (PPV) 75% and negative predictive value (NPV) 74%. The low sensitivity indicates the need for gastric biopsies. Staining had: sensitivity 95%, specificity 67%, PPV 84% and NPV 89%, with significant difference for staining (P0.05). The typical endoscopic appearance of intestinal metaplasia always coincided with staining and histology. Dysplasia was found in 0.53% ofcases, gastric cancer in 1.8% and Helicobacter pylori in 56%.Directed biopsy staining is superior to those taken at random for diagnosing metaplasia. Metaplasia was not found when endoscopy was normal and typical endoscopic appearance of metaplasia agreed with staining and histology. more...

The BISAP score is a simple system, which englobes clinical features (laboratory and imagenology tests) allowing to predict the mortality in acute pancreatitis within the first 24 hours of hospitalization.To determine the validity of the BISAP score in the prediction of prognosis and severity of acute pancreatitis.In order to validate the BISAP score, a study was performed in 57 patients with a diagnosis of acute pancreatitis at the moment of admission. The reference test was the Atlanta criteria which confirmed severe clinical course beyond the fifth day, in many cases supported by CT.71.9% were women. The mean age was 45.33 years. Biliary lithiasis was the most frequent cause (66.7%). According to the reference test 71.9% were mild and 28.1% severe. According to the BISAP score 77.2% mild and 22.8% severe. The sensitivity and specificity for the BISAP score was of 75% and 97.56% respectively. The predictive positive value was 92.31% and the predictive negative value was 90.91%.The BISAP score allows to predict the severity of the acute pancreatitis. more...