Kaempferol in ameliorating diabetes-induced fibrosis and renal damage: An in vitro and in vivo study in diabetic nephropathy mice model

Phytomedicine ◽  
2020 ◽  
Vol 76 ◽  
pp. 153235 ◽  
Author(s):  
Dilip Sharma ◽  
Rakesh Kumar Tekade ◽  
Kiran Kalia
Keyword(s):  
Diabetic Nephropathy ◽  
Renal Damage ◽  
In Vivo Study ◽  
Mice Model

P0972INHIBITION OF LYSINE63 UBIQUITINATION PREVENTS THE PROGRESSION OF RENAL FIBROSIS IN DIABETIC NEPHROPATHY IN VITRO AND IN VIVO

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Francesca Conserva ◽  
Paola Pontrelli ◽  
Rossella Menghini ◽  
Michele Rossini ◽  
Alessandra Stasi ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Drug Combination ◽  
Renal Fibrosis ◽  
Renal Damage ◽  
Mesenchymal Transition ◽  
Ubiquitinated Proteins ◽  
Collagen Iii ◽  
Sirius Red

Abstract Background and Aims Diabetic Nephropathy (DN) is the primary cause of end stage renal disease (ESRD). Our group demonstrated that in DN an accumulation of lysine63 (K63)-ubiquitinated proteins at tubular level is involved in the progression of renal damage, in particular renal fibrosis. Current treatments do not provide complete renoprotection and targeted therapies that prevent fibrosis or delay its progression are still lacking. Aim of the present study was to evaluate the renoprotective effect of specific drug and their combinations, including an inhibitor of K63 ubiquitination (K63Ub) and/or an anti-hypertensive agent, in vitro and in vivo in a murine model of DN. Method Renal Proximal Tubule Epithelial Cells (HK2) were pre-incubated with a specific inhibitor of K63Ub and/or with the ACE-inhibitor Ramipril. Accumulation of K63 ubiquitinated proteins along with α-sma expression, indicator of epithelial-to-mesenchymal transition (EMT), were analyzed through immunofluorescence and western blotting. The same drug combination was also tested in streptozotocin (STZ)-treated DBA/2J mice, a model of human DN. In mice, K63Ub was evaluated by IHC, while renal fibrosis was evaluated by Sirius red and Collagen III expression. Urinary albuminuria was measured by ELISA. Results We observed that the association of the specific K63Ub inhibitor with Ramipril was able to block hyperglycemia-induced EMT in HK2 cells by significantly reducing α-sma expression, when compared to single drugs alone (p<0.05).To demonstrate the efficacy of these drug combinations in reducing the progression of renal damage in DN we firstly confirmed the increased accumulation of K63 Ub proteins in DBA/2J STZ-treated mice (p=0.01). Interestingly, increased K63Ub in diabetic mice was also associated to increased tubular-interstitial fibrosis (p<0.05). Treatment of STZ-mice with the specific K63Ub inhibitor was able to reduce both K63Ub proteins accumulation and renal fibrosis, evaluated on kidney samples by IHC against Collagen III (p≤0.05) and by Sirius Red staining (p≤0.05) when compared to both untreated mice and mice treated with ramipril. Importantly, treatment with the K63Ub inhibitor alone did not reduce albuminuria (STZ-mice: 561.29±390.56; STZ+K63Ubinhibitor: 724.25±690.89; p=n.s.), while the drug combination including the specific K63Ub inhibitor and Ramipril, significantly reduced both K63Ub-related fibrosis and albuminuria (p=0.01), demonstrating an addictive and synergic effect of these molecules when used in combination. Conclusion Our data demonstrated and confirmed the importance of K63Ub in the progression of renal fibrosis in vitro and in vivo. We proposed and patented a novel combination of drugs that ameliorates both fibrosis and proteinuria in DN. Novel treatment regimens could represent an important goal for reducing the incidence of ESRD related to diabetes complication.

scholarly journals SO025DCR2 ACCELERATES RENAL FIBROSIS AND TUBULAR CELL SENESCENCE BY PHOSPHORYLATING TYR 194 OF PRDX1 IN DIABETIC NEPHROPATHY

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Jia Chen ◽  
Yani He
Keyword(s):  
Diabetic Nephropathy ◽  
Peroxidase Activity ◽  
Renal Fibrosis ◽  
Underlying Mechanism ◽  
Cell Senescence ◽  
Tubular Cell ◽  
Mice Model ◽  
Renal Tubulointerstitial Fibrosis

Abstract Background and Aims Cell senescence of renal tubular epithelial cells (RTECs), which is involved in renal tubulointerstitial fibrosis (TIF), is a key event in the progression of diabetic nephropathy (DN). However, the underlying mechanism remains unclear. This study aims to investigate the role and mechanism of decoy receptor 2 (DcR2) in TIF and cell senescence of RTECs. Method Streptozotocin (STZ)-induced DN mice model and high glucose (HG)-induced cell senescent model were constructed. The expression of DcR2 were regulated by ultrasound-mediated gene transfer of DcR2-siRNA or DcR2-overexpression plasmid in vivo. The co-expression of DcR2 with senescent markers (p16, p21 and SA-β-gal) and fibrotic markers (FN, collagen I and α-SMA). The interaction of DcR2 and its ligand TRAIL and antagonistic receptor DR5 were detected by laser confocal and Co-IP. The DcR2-interaction proteins were screened in renal tissue of DN patients and HG-induced cells by Co-IP combining with LC-MS/MS. The interaction of DcR2 and PRDX1 was validated by Co-IP, pull down assay and immunofluorescence co-staining. Peroxidase activity of PRDX1 was assessed by the kits of reactive oxygen species (ROS) and specific 2-cys peroxidase activity. The level of PRDX1 phosphorylation was measured through WB and the phosphorylated site were studied by wild-type PRDX1 and mutant Tyr194 PRDX1. Results DcR2 was co-expressed with senescent markers and co-localization with fibrotic markers in DN patients and STZ-DN mice. Knockdown of DcR2 effectively decreased renal fibrosis and tubular cell senescence in STZ-induced DN mice. However, DcR2 overexpression showed the opposite effects. DcR2 didn’t interacted with TRAIL or DR5 in vivo and vitro. The following study indicated that PRDX1 is a novel DcR2-interacting protein using quantitative proteomics, which have peroxidase and antioxidant activity in cytoplasm. The interaction of DcR2-PRDX1 mediated RTEC senescence in vitro. Knockdown of DcR2 inhibited the level of ROS and promoted the peroxidase activity of PRDX1, resulting in oxidative stress and cell senescence, and DcR2 overexpression presented the contrary results. DcR2 regulated the level of PRDX1 phosphorylation by interacting Tyr194 of PRDX1, and the mutant Tyr194 of PRDX1 can’t be phosphorylated by DcR2. Conclusion DcR2 mediates renal fibrosis and RTEC senescence by phosphorylating with PRDX1, suggesting DcR2 could serve as a potential therapeutic target for the amelioration of DN progression.

scholarly journals Atrasentan increased the expression of klotho by mediating miR-199b-5p and prevented renal tubular injury in diabetic nephropathy

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Wen-Ling Kang ◽  
Gao-Si Xu
Keyword(s):  
Diabetic Nephropathy ◽  
High Glucose ◽  
Tubular Injury ◽  
Mice Model ◽  
Renal Tubular Cells ◽  
Proximal Tubular ◽  
Renal Proximal Tubular ◽  
Renal Tubular

Abstract Atrasentan is a promising therapy for treating diabetic nephropathy (DN). Here we evaluated whether atrasentan down-regulated the miR-199b-5p expression, thereby increasing klotho and preventing renal tubular injury in DN. One-hundred patients with type 2 diabetes mellitus (T2DM) and 40 healthy subjects were included. A DN mice model was established by an injection of streptozotocin (STZ). Human renal proximal tubular epithelial HK-2 cells were exposed to high glucose (20 mmol/L). Treated the mice and HK-2 cells with atrasentan and we then investigated whether and how miR-199b-5p and Klotho were involved in preventing renal tubular injury in DN. In patients, the serum miR-199b-5p level increased and the klotho concentration decreased in accordance with elevated albuminuria. Atrasentan down-regulated miR-199b-5p and up-regulated klotho of the DN mice and HK-2 cells exposed to high glucose. High glucose promoted the binding of histone H3 to the miR-199b-5p promoter and atrasentan canceled this effect. MiR-199b-5p targeted the 3′ UTR of klotho. Overexpression of miR-199b-5p canceled the effects of atrasentan on klotho expression and apoptosis of renal tubular cells in both in vivo and in vitro. The increased serum klotho, mediated by miR-199b-5p, is a possible mechanism by which atrasentan prevents renal tubular injury in DN.

The Natural Course of Defibrination Syndrome Caused by Echis Colorata Venom in Man

1974 ◽  
Vol 31 (03) ◽  
pp. 420-428 ◽  
Author(s):  
M Fainaru ◽  
S Eisenberg ◽  
N Manny ◽  
C Hershko
Keyword(s):  
Factor Viii ◽  
Blood Coagulation ◽  
Major Bleeding ◽  
Renal Damage ◽  
Specific Treatment ◽  
Natural Course ◽  
Factor V ◽  
Thrombocyte Count

SummaryThe natural course of defibrination syndrome caused by Echis colorata venom (ECV) in five patients is reported. All patients developed afibrinogenemia within six hours after the bite. Concomitantly a depression in factor V was recorded. Factor VIII and thrombocyte count in blood were normal in most patients. In the light of the known effects of ECV on blood coagulation in vivo and in vitro it is concluded that the afibrinogenemia is due to intravascular clotting.Four patients had transient renal damage, manifested by oliguria, azotemia, albuminuria and cylindruria, ascribed to microthrombi in the renal glomeruli.After the bite, the natural course was benign, no major bleeding was observed, and all signs of coagulopathy reverted to normal within 7 days. Therefore we recommend no specific treatment for this condition. In the case of heavily bleeding patients, administration of antiserum against ECV and/or heparin should be considered.

Short-Chain Fatty Acids Prevent Diabetic Nephropathy In Vivo and In Vitro

Diabetes ◽  
2018 ◽  
Vol 67 (Supplement 1) ◽  
pp. 92-OR ◽  
Author(s):  
WEI HUANG ◽  
YONG XU ◽  
YOUHUA XU ◽  
LUPING ZHOU ◽  
CHENLIN GAO
Keyword(s):  
Fatty Acids ◽  
Diabetic Nephropathy ◽  
Short Chain Fatty Acids ◽  
Short Chain ◽  
Chain Fatty Acids

Formulation and Evaluation of Atorvastatin Calcium-Poly-ε-Caprolactone Nanoparticles Loaded Ocular Inserts for Sustained Release and Antiinflammatory Efficacy

2020 ◽  
Vol 21 (15) ◽  
pp. 1688-1698
Author(s):  
Germeen N.S. Girgis
Keyword(s):  
Sustained Release ◽  
In Vitro Release ◽  
Pluronic F127 ◽  
In Vivo Study ◽  
Average Weight ◽  
Drug Release Profile ◽  
Atorvastatin Calcium ◽  
Anti Inflammatory

Purpose: The work was performed to investigate the feasibility of preparing ocular inserts loaded with Poly-ε-Caprolactone (PCL) nanoparticles as a sustained ocular delivery system. Methods: First, Atorvastatin Calcium-Poly-ε-Caprolactone (ATC-PCL) nanoparticles were prepared and characterized. Then, the optimized nanoparticles were loaded within inserts formulated with Methylcellulose (MC) and Polyvinyl Alcohol (PVA) by a solvent casting technique and evaluated physically, for in-vitro drug release profile. Finally, an in-vivo study was performed on the selected formulation to prove non-irritability and sustained ocular anti-inflammatory efficacy compared with free drug-loaded ocuserts. Results: The results revealed (ATC-PCL) nanoparticles prepared with 0.5% pluronic F127 were optimized with 181.72±3.6 nm particle size, 0.12±0.02 (PDI) analysis, -27.4± 0.69 mV zeta potential and 62.41%±4.7% entrapment efficiency. Nanoparticles loaded ocuserts manifested compatibility between drug and formulation polymers. Moreover, formulations complied with average weight 0.055±0.002 to 0.143±0.023 mg, and accepted pH. ATC-PCL nanoparticles loaded inserts prepared by 5% MC showed more sustained, prolonged in-vitro release over 24h. In-vivo study emphasized non-irritability, ocular anti-inflammatory effectiveness represented by smaller lid closure scores, and statistically significant lowering in PMN count after 3h. Conclusion: These findings proposed a possibly simple, new and affordable price technique to prepare promising (ATC-PCL) nanoparticles loaded inserts to achieve sustained release with prolonged antiinflammatory efficacy.

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