Site specific regulation in the kidney of endothelin and its receptor subtypes by cyclosporine

Site selective regulation in the kidney of endothelin and its receptor subtypes by cyclosporine. Endothelin (Et) has been suggested by us and others to play a role in glomerular dysfunction that characterizes cyclosporine (Cs)-associated nephrotoxicity. Since Et exerts its actions through at least two receptor subtypes, and because these receptor subtypes have particular distributions in the renal parenchyma, we investigated changes in mRNA expression for Et and its receptor subtypes in glomeruli and medulla of rats treated with Cs. Polymerase chain reaction coupled with reverse transcription (RT-PCR) method was used to assess prepro-Et-1, type A (EtA) and type B (EtB) receptor mRNA at 1, 3, 6, and 24 hours after Cs (20 mg/kg body wt i.v.). Results were normalized to the expression of β-actin as an internal standard. Compared with control rats, glomerular mRNA expression for prepro-Et-1 was not affected by Cs. Similarly, Cs did not significantly change the glomerular mRNA expression of either EtA or EtB receptor subtypes. By contrast, in the medulla there was a marked and persistent increase in the expression for prepro-Et-1 and the EtB receptor subtype: prepro-Et-1 at 1, 3, 6, and 24 hours was 336 ± 61, 295 ± 65, 339 ± 73,440 ± 123% of controls, respectively (P < 0.05 compared with controls at each time point). The EtB receptor mRNA at 1, 3, 6, 24 hours was 164 ± 22, 157 ± 15, 148 ± 14, 116 ± 18% (compared with controls, P < 0.01 at 3hr and P < 0.05 at 1 and 6 hr), while the mRNA expression for EtA was not affected by Cs treatment. These results demonstrate that, in vivo, Cs selectively modulates renal mRNA expression for Et peptide and one of its receptor subtypes. Furthermore, the modulation is site specific. These changes are most conspicuous in the renal medulla, such that mRNA expression for prepro-Et-1 and EtB increases and remains elevated for at least six hours after Cs administration. These alterations may contribute to the vasospastic as well as proliferative abnormalities which characterize Cs nephrotoxicity

Strategic locus for the activation of the superoxide dismutase gene in the nephron

Strategic locus for the activation of the superoxide dismutase gene in the nephron. Upon exposure to a transient ischemia, the distal tubule of the kidney often escapes the severe damage which afflicts the proximal tubule. To ascertain whether this feature of the distal tubule is attributable to its intrinsic cellular properties, we focused on two pairs of unique tubule segments; distal versus proximal convoluted tubules in the superficial cortex and distal versus proximal straight tubules in the outer stripe of the outer medulla. These tubules were chosen because, firstly, they can be identified by morphology and immunostaining, and secondly, each pair has the same anatomical relationship to the circulation. Detailed morphometric analyses were performed six hours following unilateral transient ischemia in adult rats to semiquantitate the local tissue damage in these specific nephron segments. The architecture of the distal convoluted and straight tubules was remarkably well preserved, contrasting to the moderate to extensive necrotic changes seen in the proximal tubules. In search of the potential intrinsic cellular mechanism that underlies the observed difference, we examined the segmental distribution along the nephron of manganese superoxide dismutase gene transcripts by in situ hybridization. This antioxidant enzyme gene was expressed primarily in the distal tubules with contrastingly low levels of expression in the proximal tubules. Moreover, following ischemia-reperfusion, this distal tubule-dominant pattern was further accentuated immediately following reperfusion. The study indicates that the marked difference between the proximal and distal tubules in their susceptibility to injury in vivo is attributable to their intrinsic cellular properties, which include the local level of antioxidants

Mineralocorticoid Receptor Blocker Protects against Podocyte-Dependent Glomerulosclerosis

Background: We previously showed that angiotensin type 1 receptor (AT1) blocker (ARB) attenuates glomerular injury in Nphs1-hCD25 (NEP25) transgenic mice, a model of selective podocyte injury. However, subsequent studies in NEP25 mice with podocyte-specific deficiency of AT1 revealed that the protective effects of ARB are not through the podocyte AT1, thereby raising the possibility that the protective effects of ARB involve mineralocorticoids. Methods: NEP25 mice were treated with the mineralocorticoid receptor blocker (MRB) spironolactone (25 mg/kg/day, n = 10), the ARB losartan (250 mg/kg/day, n = 11), both (ARB+MRB, n = 8) or vehicle (Vehicle, n = 9) from day –7 to day 9 of induction of podocyte injury. Results: Although MRB did not reduce systolic blood pressure or proteinuria, addition of MRB to ARB significantly attenuated glomerulosclerosis (glomerulosclerosis index: ARB+MRB 1.67 ± 0.19 vs. MRB 2.01 ± 0.29, ARB 2.35 ± 0.19, and Vehicle 2.25 ± 0.26, p Conclusion: These data suggest that, while MRB does not attenuate proteinuria caused by podocyte-specific injury, it provides protective effects against glomerulosclerosis that is independent of systemic blood pressure

Oxidants induce transcriptional activation of manganese superoxide dismutase in glomerular cells

Oxidants induce transcriptional activation of manganese superoxide dismutase in glomerular cells. Cultured rat glomerular mesangial and epithelial cells and bovine glomerular endothelial cells were exposed to various concentrations of hydrogen peroxide (H2O2). Mesangial cells treated with 10 to 100 µM H2O2 for 24 hours showed a two- to ninefold increase in Mn-SOD mRNA expression associated with significantly (P < 0.005) increased Mn-SOD activity (22.2 ± 1.2 and 12.2 ± 0.7 µ/mg protein for H2O2 100 µM treated and untreated cells, respectively). In contrast, expression of Cu-Zn SOD and β-actin mRNA was not affected by H2O2. Induction of Mn-SOD mRNA by H2O2 was inhibited by actinomycin-D (4 µM) treatment. Glomerular endothelial cells also showed an increase in Mn-SOD mRNA expression following 100 µM H2O2 treatment, as did glomerular epithelial cells following treatment with 500 and 1000 µM H2O2 but not with 100 µM. Transcriptional activity of the Mn-SOD gene was assessed with a fusion reporter gene consisting of a luciferase gene (pGL2P) and a 1.2 kb fragment from the rat Mn-SOD genomic DNA (-806 to +408 bp of the transcription initiation site, -806:+408). The construct was transfected into rat glomerular mesangial and epithelial cells. Mesangial and epithelial cells transfected with pGL2P (-806:+408) and treated with H2O2 (100 µM and 1 mM for mesangial and epithelial cells, respectively) demonstrated some threefold increase in luciferase activity, whereas cells transfected with pGL2P lacking the Mn-SOD fragment did not show changes in luciferase activity following H2O2 treatment. Other oxidants, namely α- and β-naphthoflavone (50 µM to mesangial cells) and puromycin aminonucleoside (25 to 50 µg/ml to epithelial cells), also induced transcriptional activation (2- to 5-fold increase) in these cells. Thus, Mn-SOD levels in glomerular cells are enhanced when they are exposed to oxidant stress, and this up-regulation involves transcriptional activation. Further, the Mn-SOD gene contains enhancer element(s) which respond to diverse oxidant stress. The inducibility by oxidants of local Mn-SOD demonstrates that glomerular SOD may play a decisive role in the pathogenesis of glomerular injuries in which the balance between oxidants and antioxidants is critical

In Vivo Model of Osteoarthritis to Compare Allogenic Amniotic Epithelial Stem Cells and Autologous Adipose Derived Cells

SIMPLE SUMMARY: An early resolution of osteoarthritis (OA), through minimally invasive orthobiological solutions, would be important to enable a return to daily and sport activities, and delay prosthesis solutions. No study has yet evaluated amniotic epithelial stem cells (AECs) in OA. They could be considered a valid alternative to adipose derived cells, expanded or concentrated, because they differentiate into three lineages and express mesenchymal and embryonic markers, without a tumorigenic phenotype. The innovative aspects of this study are the comparison of three injective orthobiological treatments, the in vivo use of AECs in OA, and the evaluation of structural and inflammatory fronts of OA for up to six months. ABSTRACT: The challenge of osteoarthritis (OA) is to find a minimally invasive orthobiological therapy to contrast OA progression, on inflammatory and structural fronts. The aim of the present study is to compare the effects of an intra-articular injection of three orthobiological treatments, autologous culture expanded adipose-derived mesenchymal stromal cells (ADSCs), autologous stromal vascular fraction (SVF) and allogenic culture expanded amniotic epithelial stem cells (AECs), in an animal model of OA. OA was induced in 24 sheep by bilateral lateral meniscectomy and, at 3 and 6 months post-treatment, the results were analyzed with macroscopy, histology, histomorphometry, and biochemistry. All the three treatments showed better results than control (injection of NaCl), but SVF and AECs showed superiority over ADSCs, because they induced higher cartilage regeneration and lower inflammation. SVF showed better results than AECs at 3 and 6 months. To conclude, SVF seems to be more favorable than the other biological options, because it is easily obtained and rapidly used after harvesting, with good healing potential. AECs cause no discomfort and could be also considered for the treatment of OA joints

Preparation method and growth factor content of platelet concentrate influence the osteogenic differentiation of bone marrow stromal cells

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17)脊椎々間板手術症例の検討 : とくに椎間板障害例における手術適応について(第399回千葉医学会整形外科例会,第8回千葉整形外科災害外科臨床懇談会,千葉県労災指定医集談会)

<p>Data are expressed as medians and interquartile ranges; comparisons among time points, as determined by Friedman-ANOVA test, and between L-PRP and HA treatments, as determined by the Mann-Whitney U test, are not significant. w = week, m = month</p

Design and baseline characteristics of the finerenone in reducing cardiovascular mortality and morbidity in diabetic kidney disease trial

Background: Among people with diabetes, those with kidney disease have exceptionally high rates of cardiovascular (CV) morbidity and mortality and progression of their underlying kidney disease. Finerenone is a novel, nonsteroidal, selective mineralocorticoid receptor antagonist that has shown to reduce albuminuria in type 2 diabetes (T2D) patients with chronic kidney disease (CKD) while revealing only a low risk of hyperkalemia. However, the effect of finerenone on CV and renal outcomes has not yet been investigated in long-term trials. Patients and Methods: The Finerenone in Reducing CV Mortality and Morbidity in Diabetic Kidney Disease (FIGARO-DKD) trial aims to assess the efficacy and safety of finerenone compared to placebo at reducing clinically important CV and renal outcomes in T2D patients with CKD. FIGARO-DKD is a randomized, double-blind, placebo-controlled, parallel-group, event-driven trial running in 47 countries with an expected duration of approximately 6 years. FIGARO-DKD randomized 7,437 patients with an estimated glomerular filtration rate >= 25 mL/min/1.73 m(2) and albuminuria (urinary albumin-to-creatinine ratio >= 30 to <= 5,000 mg/g). The study has at least 90% power to detect a 20% reduction in the risk of the primary outcome (overall two-sided significance level alpha = 0.05), the composite of time to first occurrence of CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure. Conclusions: FIGARO-DKD will determine whether an optimally treated cohort of T2D patients with CKD at high risk of CV and renal events will experience cardiorenal benefits with the addition of finerenone to their treatment regimen. Trial Registration: EudraCT number: 2015-000950-39; ClinicalTrials.gov identifier: NCT02545049