BackgroundIndoleamine 2, 3-dioxygenase (IDO) is an immunomodulatory molecule that has been implicated in several biological processes. Although IDO has been linked with some renal diseases, its role in renal fibrosis is still unclear. Because IDO may be modulated by TGF-β1, a potent fibrogenic molecule, we hypothesized that IDO could be involved in renal fibrosis, especially acting in the TGF-β1-induced tubular epithelial-mesenchymal transition (EMT). We analyzed the IDO expression and activity in a model of renal fibrogenesis, and the effect of the IDO inhibitor 1-methyl-tryptophan (MT) on TGF-β1-induced EMT using tubular cell culture.MethodsMale Wistar rats where submited to 7 days of UUO. Non-obstructed kidneys (CL) and kidneys from SHAM rats were used as controls. Masson’s Tricrome and macrophages counting were used to chatacterize the tissue fibrosis. The EMT was analysed though immunohistochemistry and qRT-PCR. Immunohistochemestry in tissue has used to show IDO expression.MDCK cells were incubated with TGF- β1 to analyse IDO expression. Additionally, effects of TGF- β1 and the inhibition of IDO over the EMT process was acessed by immunoessays and scrath wound essay.ResultsIDO was markedly expressed in cortical and medular tubules of the UUO kidneys. Similarly to the immunolocalizaton of TGF- β1, accompanied by loss of e-cadherin expression and an increase of mesenchymal markers. Results in vitro with MDCK cells, showed that IDO was increased after stimulus with TGF-β1, and treatment with MT potentiated its expression. MDCK stimulated with TGF-β1 had higher migratory activity (scratch-wound assay), which was exacerbated by MT treatment.ConclusionsIDO is constitutively expressed in tubular cells and increases during renal fibrogenesis. Although IDO is induced by TGF-β1 in tubular cells, its chemical inhibitor acts as a profibrotic agent.

Women with preeclampsia (PE) form a vulnerable group for vitamin D3 deficiency. Reabsorption of vitamin D3 occurs in the proximal tubule after being endocytosed in combination with DBP (vitamin D binding protein) by the megalin/cubilin receptor. Because proteinuria promotes tubule injury and dysfunction, we hypothesized that the proteinuria present in pe could promote the loss of these components into the urine. Twenty preeclamptic patients and ten normal pregnant women with a gestational age greater than 20 weeks composed three groups: NC, normotensive control pregnant patients; PE, non-proteinuric preeclamptic patients; and PPE, preeclamptic patients with proteinuria. When proteinuria was absent, preeclampsia was diagnosed accordingly to the American college of Obstetricians and Gynecologists' (ACOG) guideline. The presence of 24-hour proteinuria equal to or greater than 300 mg was considered to form the PPE group. Urinary cubilin, megalin, and DBP were measured by ELISA and normalized by urinary creatinine. Regarding gestational age, there was no difference between the groups. NC group had arterial pressure within normal values, whereas PE and PPE groups had a significant increase (p < 0.01). As expected, PPE group presented elevated ACR (p < 0.05), accompanied by large amounts of cubilin and DBP in the urine (p < 0.05 vs. NC and PE). No difference was found in urinary megalin. PPE patients showed more chance of shedding cubilin into the urine compared to non-proteinuric patients (odds ratio 12.7 (1.2-136.3). In conclusion, this study further tightens the relationship between pe and vitamin D 3 deficiency, since proteinuria present in pe induces the loss of molecules responsible for renal tubular vitamin D 3 reabsorption for subsequent activation. Combined with other factors, the proteinuria may intensify vitamin D 3 deficiency in PE. Preeclampsia (PE) is a multisystem disorder of pregnancy in which renal damage has a significant contribution. A set level of proteinuria has been used to predict high-risk groups and define treatment for PE. In terms of physiopathology, proteinuria emerges due to processes that disturb the glomerular filtration barrier. Besides the classical mechanisms of glomerular damage such as endotheliosis and loss of podocytes 1,2 , an evidence for tubular injury have also been reported. Markers of proximal tubule injury such as kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), and retinol-binding protein (RBP) were higher in the urine of PE patients than in urine of healthy pregnancies 3 In addition, urinary lysosomal enzyme excretion is elevated in patients with PE compared to a pregnant woman with chronic hypertension 4. Burwick et al. demonstrated that pregnancies affected by severe PE have high amounts of KIM-1 in the urine, which was closely correlated with urinary complement activation products, suggesting that tubular injury in PE is intensified by inflammation-activated complement 5. Direct effects of proteinuria on tubular cell damage have been ...

Gestational hypothyroidism has 5.1% prevalence and is associated with severe consequences as hypertensive complications and preeclampsia (PE). Based on the higher mortality associated with PE, the objective of this study is to identify new biomarkers useful as prognostic and therapeutically parameters. Methodology: The study was approved by the IRB Number: 665331/679727. We selected three volunteers to each group and constructed libraries of circulating RNA, for healthy pregnant women (HPW), preeclampsia (PEC) and gestational hypothyroidism (GHT). We collected blood samples for TSH and Free T4 (FT4) measurement and a PAX Gene Tube for RNA analysis. RNA extraction was performed by the PAXgene Blood RNA extraction kit (Qiagen NL, DE). NGS platform, Ion Proton System was used with Ion AmpliSeq Gene human transcriptome (Thermo Fisher Scientific Manufacturer) kit to construct the transcriptome libraries. The R software platform version 3.4.1 (R Foundation for Statistical Computing, Vienna, Austria. URL: R-project.org/) with the edgeR package 3.16.5 were used to determine the differentially expressed genes (DEG). We compared our GHT DEG with plasma transcriptome libraries from healthy pregnant women in the second and third trimesters (access number: GSE56899) available in GEO Dataset (ncbi.nlm.nih.gov/geo) using the limma package 3.26.8, p <0,05. We also used Random Forest (RF) analysis to rank the variables, Spearman Correlation, and multiclass ROC. Results: We encountered 713 DEG in the HPW with matrix metalloproteinases (MMP) upregulated, MMP8 (logFC 4.91) and MMP9 (logFC 3.44). In GEO Dataset we also found the MMP8 upregulated. We detected 195 DEG in PEC with MMP9 downregulated (logFC -2.49) and 745 DEG in GHT with MMP8 (logFC -4.06) and MMP9 (logFC -3.26) downregulated. We obtained 571 DEG in the analysis obtained by GEO Dataset and 26 genes are in common with our GHT DEG, including the MMP8 . Between the 26 in common, 88.46% of genes were downregulated in GHT and 92.30% were upregulated GEO Dataset. We found a negative correlation of MMP8 with systolic blood pressure (SBP) and diastolic blood pressure (DBP) (r: -0.313; r: -0.285), MMP9 with SBP and DBP (r: -0.349; r: -0.384), MMP8 and MMP9 with weight (r: -0.223; r: -0.209) and MMP9 with TSH (r: 0.633). From our RF model, we selected the variables SBP and DBP, FT4 and weight that is possibly modified by the difference in expression of MMP8 and MMP9 . We obtained an area under the curve of 0.97 for TSH, weight and those genes. Conclusion: Several metalloproteinases are involved in placental development, implantation and angiogenesis. We observed ...