Experimental model of nephropathy associated with diabetes mellitus in mice

Purpose: Nontransmissible chronic diseases, such as diabetes mellitus (DM) and nephropathy, affect a significant portion of the population, often treated due to injuries that require healing and regeneration. To create an experimental model of associated comorbidities, for healing and regeneration studies, protocols for induction of nephropathy by ischemia and reperfusion (I/R) and induction of DM by injection of streptozotocin (STZ) were associated. Methods: Sixty-four mice (Mus musculus), female, adult, Swiss strain, weighing approximately 20 g, were divided into four groups: G1: control (n = 24), G2: nephropathy group (N) (n = 7), G3, DM (n = 9), and G4: N+DM (n = 24). Arteriovenous stenosis (I/R) of the left kidney was performed as the first protocol. The animals received a hyperlipidemic diet for 7 days after the injection of STZ (150 mg/kg, via i.p.) and an aqueous glucose solution (10%) for 24 h. The animals in the G3 and G4 groups were observed for 14 days before receiving the diet and STZ. The evolution of nephropathy was observed using a urine test strip and the DM, through the analysis of blood glucose with a reagent strip on a digital monitor. Results: The ischemic induction protocols of nephropathy and DM with STZ, associated, were sustainable, low-cost, and without deaths. There were alterations compatible with initial renal alterations, in the first 14 days, such as increased urinary density, pH alteration, presence of glucose, proteins and leukocytes, when compared to the control group. DM was confirmed by the presence of hyperglycemia 7 days after induction and its evolution after 14 days. The animals in the G4 group showed constant weight loss when compared to the other groups. It was possible to observe morphological alterations in the kidneys submitted to I/R, regarding coloration, during surgery and after the end of the observation period, in the volume and size of the left kidney, when compared to the contralateral kidney. Conclusion: It was possible to induce nephropathy and DM associated in the same animal, in a simple way, confirmed with rapid tests, without losses, providing a basis for future studies.

Amelioration of exosome and mesenchymal stem cells in rats infected with diabetic nephropathy by attenuating early markers and aquaporin-1 expression / Melhoria das células-tronco exossomais e mesenquimais em ratos infectados com nefropatia diabética atenuando marcadores precoces e expressão de aquaporina-1

Diabetic nephropathy (DN) is a prevalent diabetic microvascular condition. It is the leading cause of kidney disease in the advanced stages. There is no currently effective treatment available. This research aimed to investigate the curative potentials of exosomes isolated from mesenchymal stem cells affecting DN. This study was performed on 70 male adult albino rats. Adult rats were randomized into seven groups: Group I: Negative control group, Group II: DN group, Group III: Balanites treated group, Group IV: MSCs treated group, Group V: Exosome treated group, Group VI: Balanites + MSCs treated group and Group VII: Balanites + exosome treated group. Following the trial period, blood and renal tissues were subjected to biochemical, gene expression analyses, and histopathological examinations. Results showed that MDA was substantially increased, whereas TAC was significantly decreased in the kidney in the DN group compared to normal health rats. Undesired elevated values of MDA levels and a decrease in TAC were substantially ameliorated in groups co-administered Balanites aegyptiacae with MSCs or exosomes compared to the DN group. A substantial elevation in TNF-α and substantially diminished concentration of IGF-1 were noticed in DN rats compared to normal health rats. Compared to the DN group, the co-administration of Balanites aegyptiacae with MSCs or exosomes substantially improved the undesirable elevated values of TNF-α and IGF-1. Furthermore, in the DN group, the mRNA expression of Vanin-1, Nephrin, and collagen IV was significantly higher than in normal healthy rats. Compared with DN rats, Vanin-1, Nephrin, and collagen IV Upregulation were substantially reduced in groups co-administered Balanites aegyptiacae with MSCs or exosomes. In DN rats, AQP1 expression was significantly lower than in normal healthy rats. Furthermore, the groups co-administered Balanites aegyptiacae with MSCs or exosomes demonstrated a substantial increase in AQP1 mRNA expression compared to DN rats.

A nefropatia diabética (ND) é uma condição microvascular diabética prevalente. É a principal causa de doença renal em estágios avançados. Atualmente, não há tratamento eficaz disponível. Esta pesquisa teve como objetivo investigar os potenciais curativos de exossomos isolados de células-tronco mesenquimais que afetam a ND. Este estudo foi realizado em 70 ratos albinos adultos machos. Ratos adultos foram randomizados em sete grupos: Grupo I: Grupo de controle negativo, Grupo II: Grupo DN, Grupo III: Grupo tratado com Balanites, Grupo IV: Grupo tratado com MSCs, Grupo V: Grupo tratado com exossomos, Grupo VI: Grupo tratado com Balanites + MSCs e Grupo VII: Balanites + grupo tratado com exossomas. Após o período experimental, o sangue e os tecidos renais foram submetidos a análises bioquímicas, de expressão gênica e exames histopatológicos. Os resultados mostraram que o MDA aumentou substancialmente, enquanto o TAC diminuiu significativamente no rim no grupo DN em comparação com ratos saudáveis normais. Valores elevados indesejados de níveis de MDA e uma diminuição no TAC foram substancialmente melhorados em grupos coadministrados Balanites aegyptiacae com MSCs ou exossomas em comparação com o grupo DN. Uma elevação substancial em TNF-α e uma concentração substancialmente diminuída de IGF-1 foram observadas em ratos DN em comparação com ratos saudáveis normais. Em comparação com o grupo DN, a coadministração de Balanites aegyptiacae com MSCs ou exossomas melhorou substancialmente os valores elevados indesejáveis de TNF-α e IGF-1. Além disso, no grupo DN a expressão de mRNA de vanina-1, nefrina e colágeno IV foi significativamente maior do que em ratos saudáveis normais. Comparado com ratos DN, Vanin-1, Nephrin e colágeno IV Upregulation foram substancialmente reduzidos em grupos co-administrados Balanites aegyptiacae com MSCs ou exossomos. Em ratos DN, a expressão de AQP1 foi significativamente menor do que em ratos saudáveis normais. Além disso, os grupos que coadministraram Balanites aegyptiacae com MSCs ou exossomos demonstraram um aumento substancial na expressão de mRNA de AQP1 em comparação com ratos DN.

Genistein improves mitochondrial function and inflammatory in rats with diabetic nephropathy via inhibiting MAPK/NF-κB pathway

Purpose: To investigate the effect of genistein on inflammation and mitochondrial function of diabetic nephropathy. Methods: Diabetic nephropathy model was established in Sprague-Dawley rats. Automatic biochemical analyzer was employed to detect the kidney function index, serum creatinine, serum urea nitrogen, and 24 h-urine protein and blood glucose. Hematoxylin and eosin staining and periodic acid Schiff staining were used to observe renal morphology. Mitochondrial changes and podocyte integrity were monitored by transmission electron microscope. The expression levels of mfn2, NOX4, P53, MAPK, and NF-κB were detected by Western blotting. The changes of mitochondrial membrane potential were measured by JC-1. The level of mfn2 was assessed by immunofluorescence assay. Results: Genistein ameliorated the kidney function with reduced Scr and blood glucose. The expressions of NOX4, MAPK, p65 and p53 were downregulated, while the expression of mnf2 was the opposite in genistein-treated kidneys. Further investigations revealed that genistein reduced expansion of mesangial matrix and oxidative stress, protected podocyte integrity and increased mitochondrial membrane potential. Conclusions: Genistein could alleviate diabetic nephropathy through inhibiting MAPK/NF-κB pathway, improving mitochondrial function and anti-inflammatory.To investigate the effect of genistein on inflammation and mitochondrial function of diabetic nephropathy. Diabetic nephropathy model was established in Sprague-Dawley rats. Automatic biochemical analyzer was employed to detect the kidney function index, serum creatinine, serum urea nitrogen, and 24 h-urine protein and blood glucose. Hematoxylin and eosin staining and periodic acid Schiff staining were used to observe renal morphology. Mitochondrial changes and podocyte integrity were monitored by transmission electron microscope. The expression levels of mfn2, NOX4, P53, MAPK, and NF-κB were detected by Western blotting. The changes of mitochondrial membrane potential were measured by JC-1. The level of mfn2 was assessed by immunofluorescence assay. Genistein ameliorated the kidney function with reduced Scr and blood glucose. The expressions of NOX4, MAPK, p65 and p53 were downregulated, while the expression of mnf2 was the opposite in genistein-treated kidneys. Further investigations revealed that genistein reduced expansion of mesangial matrix and oxidative stress, protected podocyte integrity and increased mitochondrial membrane potential. Genistein could alleviate diabetic nephropathy through inhibiting MAPK/NF-κB pathway, improving mitochondrial function and anti-inflammatory.

Ramipril can alleviate the accumulation of renal mesangial matrix in rats with diabetic nephropathy by inhibiting insulin-like growth factor-1

Purpose:To investigate the impact of Ramipril (RAM) on the expressions of insulin-like growth factor-1 (IGF-1) and renal mesangial matrix (RMM) in rats with diabetic nephropathy (DN).Methods:The Sprague Dawley rats were divided into normal control (NC) group (n = 12), DN group (n = 11), and DN+RAM group (n = 12). The ratio of renal weight to body weight (RBT), fasting blood glucose (FBG), HbA1c, 24-h urine protein (TPU), blood urea nitrogen (BUN), creatinine (Cr), renal pathological changes, the levels of IGF-1, fibronectin (FN), type IV collagen (Col-IV), and matrix metalloproteinases (MMP)-2 were compared among the groups.Results:Compared with NC group, the RBT, FBG, HbA1c, TPU, BUN, Cr, and RMM in DN group were significantly increased (P < 0.05), the IGF-1, FN, and Col-IV were significantly upregulated (P < 0.05), while MMP was significantly downregulated (P < 0.05). Compared with DN group, the indexes except for the FBG and HbA1c in DN+RAM group were significantly improved (P < 0.05), among which IGF-1 exhibited significant positive correlation with TPU(r=0.937), FN(r=0.896) and Col-IV(r=0.871), while significant negative correlation with MMP-2 (r=-0.826) (P<0.05).Conclusion:RAM may protect the kidneys by suppressing IGF-1 and mitigating the accumulation of RMM.(AU)

Nefropatia diabética em cães: revisão de literatura / Diabetic nephropathy in dogs: literature review

O diabetes mellitus que se desenvolve por deficiência de insulina e hiperglicemia crônica usualmente apresenta complicações que incluem catarata, retinopatia, infecções recorrentes e cetoacidose. A hiperglicemia crônica pode promover complicações a longo prazo como a nefropatia diabética. Nos humanos, a nefropatia diabética é caracterizada principalmente por lesão glomerular e geralmente está associada a hipertensão arterial sistêmica. Dentre os mecanismos propostos para a fisiopatologia da nefropatia diabética tem sido referida a lesão glomerular, assim como a lesão nos túbulos renais. Contudo, até o momento existem poucos estudos investigando a relação entre diabetes mellitus e lesão renal em cães. Esta revisão tem por objetivos esclarecer os possíveis mecanismos fisiopatológicos da nefropatia diabética e discutir o conhecimento disponível sobre sua ocorrência em cães.

Diabetes mellitus caused by insulin deficiency and chronic hyperglycemia usually presents complications including cataract, retinopathy, recurrent infections and ketoacidosis. Chronic hyperglycemia can cause long-term complications such as diabetic nephropathy. In humans, diabetic nephropathy is characterized mainly by glomerular injury and usually it is associated with systemic hypertension. The mechanisms proposed for the pathogenesis of diabetic nephropathy could be glomerular injury, as well as renal tubular damage. However, there are few studies analyzing the relationship between diabetes mellitus and kidney damage in dogs. The objective of this review is to clarify the pathophysiological mechanisms of diabetic nephropathy and to analyze the available information about its occurrence in dogs.

Nefropatia diabética em cães: revisão de literatura / Diabetic nephropathy in dogs: literature review

O diabetes mellitus que se desenvolve por deficiência de insulina e hiperglicemia crônica usualmente apresenta complicações que incluem catarata, retinopatia, infecções recorrentes e cetoacidose. A hiperglicemia crônica pode promover complicações a longo prazo como a nefropatia diabética. Nos humanos, a nefropatia diabética é caracterizada principalmente por lesão glomerular e geralmente está associada a hipertensão arterial sistêmica. Dentre os mecanismos propostos para a fisiopatologia da nefropatia diabética tem sido referida a lesão glomerular, assim como a lesão nos túbulos renais. Contudo, até o momento existem poucos estudos investigando a relação entre diabetes mellitus e lesão renal em cães. Esta revisão tem por objetivos esclarecer os possíveis mecanismos fisiopatológicos da nefropatia diabética e discutir o conhecimento disponível sobre sua ocorrência em cães.(AU)

Diabetes mellitus caused by insulin deficiency and chronic hyperglycemia usually presents complications including cataract, retinopathy, recurrent infections and ketoacidosis. Chronic hyperglycemia can cause long-term complications such as diabetic nephropathy. In humans, diabetic nephropathy is characterized mainly by glomerular injury and usually it is associated with systemic hypertension. The mechanisms proposed for the pathogenesis of diabetic nephropathy could be glomerular injury, as well as renal tubular damage. However, there are few studies analyzing the relationship between diabetes mellitus and kidney damage in dogs. The objective of this review is to clarify the pathophysiological mechanisms of diabetic nephropathy and to analyze the available information about its occurrence in dogs.(AU)