Renal Tissues Of Diabetic Nephropathy Rats Research Articles

Radix puerariae in the treatment of diabetic nephropathy: A network pharmacology analysis and experimental validation

Abstract Radix puerariae has become the most commonly used medicine for diabetic nephropathy (DN). However, the mechanism of Radix puerariae in the treatment of DN is not completely clear. This study is to determine the active ingredients, targets, and signaling pathways of Radix puerariae for the treatment of DN using network pharmacology analysis and animal experiments to confirm its possible mechanism of action. A total of 12 potential effective components and 10 key therapeutic targets were obtained. The Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that the use of Radix puerariae in DN treatment mainly involves HIF-1 signaling pathway, TNF signaling pathway, mTOR signaling pathway, PI3K-Akt signaling pathway, FoxO signaling, and VEGF signaling pathway. Molecular docking proved that the combined activity of the components with potential key targets were excellent. Animal experiments showed that Radix puerariae could improve the renal pathological structure in DN rats. Radix puerariae could decrease the content of AKT1, IL-6, INS, and reduce the expression levels of p-Akt/Akt and p-FoxO1/FoxO1 in renal tissue of DN rats. This study provides insight into the therapeutic potential and molecular mechanisms of Radix puerariae for treating DN.

RETRACTED: BMSCs-derived exosomal microRNA-let-7a plays a protective role in diabetic nephropathy via inhibition of USP22 expression

Diabetic nephropathy (DN) is the leading cause of end-stage kidney disease. The molecular pathogenesis of DN is still poorly understood. This study was designed to investigate the protective effect of bone marrow mesenchymal stem cell (BMSCs)-derived exosome (Exo)-transported microRNA-let-7a (miR-let-7a) on DN by targeting ubiquitin-specific protease 22 (USP22). BMSCs of rats were cultured, and exosomes were identified. A rat models of DN were established in this study, and the rats were injected with Exo, Exo-let-7a mimic, or si-USP22 to figure their functions in renal function indicators blood urea nitrogen (BUN) and serum creatinine (SCr), blood lipid-related indicators total cholesterol (TC) and triglyceride (TG), renal cell apoptosis, oxidative stress, and expression of N-cadherin and vimentin in renal tissues. MiR-let-7a and USP22 targeting relationship was validated. Suppressed miR-let-7a and over-expressed USP22 exhibited in renal tissues of DN rats. Exosomes increased miR-let-7a and repressed USP22 in renal tissues of DN rats. Moreover, elevated exosomal miR-let-7a or silenced USP22 reduced SCr, BUN, TG and TC, suppressed apoptosis of renal cells and oxidative stress, and inhibited N-cadherin and vimentin expression in renal tissues of DN rats. MiR-let-7a had a targeting relationship with USP22. Our study highlights that BMSCs-derived exosomal miR-let-7a represses renal cell apoptosis, which plays a protective role in DN through down-regulation of USP22.

Liraglutide attenuates renal tubular ectopic lipid deposition in rats with diabetic nephropathy by inhibiting lipid synthesis and promoting lipolysis

Liraglutide is a new hypoglycemic drug. The previous studies have shown that liraglutide can improve the renal outcomes of patients with type 2 diabetes. Recently, it was approved by the U.S. FDA for used as a weight-loss drugs. However, the mechanism of its improvements of renal function in diabetic nephropathy patients is unclear. In addition, the effect of liraglutide on lipid metabolism is also not clear. The purpose of this study was to investigate the effects and mechanisms of liraglutide in alleviating ectopic lipid deposition (ELD) in rats with diabetic nephropathy (DN). Male Sprague-Dawley (SD) rats were treated with high-fat diet + unilateral nephrectomy + low-dose STZ combined to establish a DN rat model to evaluate the lipid-lowering effect of liraglutide. Liraglutide at 0.4 mg/kg/d were subcutaneous injected into for 12 weeks (DN + liraglutide group). After the DN rat model was established, body weight loss, 24-h urine volume increasing, serum triglycerides (TG) and serum total cholesterol (TCh) increasing, ectopic lipid droplet deposition in renal tubular increasing, mesangial proliferation in renal tissue were observed in DN rats. The treatment with liraglutide could reduce the body weight and the average daily food intake of the rats, as well as TG, TCh, and ectopic lipid droplet deposition in renal tubular. Metabolomics result showed that serum differential metabolites between the DN - vehicle control group and the DN + liraglutide group mainly included serine, threonine, phenylalanine, oxyproline, threonine, sorbitol, glyceryl monostearate, glycerol monostearate, and β-d-glucuronic acid. Moreover, liraglutide can reduce plasma lipid levels in DN rats by increasing the products of lipolysis including 1-monopalmitin and 1-monoostearin. Immunohistochemistry and Western blot showed that the expression levels of lipid synthesis-related sterol regulatory element binding protein 1 (SREBP-1) and fatty acid synthase (FAS) were significantly increased, and lipolysis-related adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) were significantly decreased both in the renal tissue of DN rats and PA-induced HK-2 cells (lipid droplet accumulation model). However, liraglutide can attenuate renal tubular ectopic lipid deposition in DN rats by inhibiting SREBP-1, FAS and increasing ATGL, HSL protein expression level, and also ameliorated PA-induced lipid accumulation in renal tubular epithelial cells. These lipid metabolism changes were attributed to liraglutide by upregulating AMP-activated protein kinase (AMPK) phosphorylation in the kidney of DN rats. Collectively, these findings confirm that liraglutide inhibits lipid synthesis and promotes lipolysis to attenuate renal ectopic lipid deposition in DN rats by promoting AMPK phosphorylation.

Regulatory Effect of 1,25(OH)2D3 on TGF-β1 and miR-130b Expression in Streptozotocin-Induced Diabetic Nephropathy in Rats.

Objective To investigate the role of microRNA-130b in 1,25(OH)2D3 mediated improvement of renal fibrosis via transforming growth factor-beta 1 in a rat model of diabetic nephropathy (DN). Methods DN was induced in 30 rats by intraperitoneal injection of streptozotocin. These rats were randomly allocated to the DN group, TGF-β1 overexpression group (in situ injection of TGF-β1 lentivirus to kidney tissues), and TGF-β1 siRNA group (in situ injection of TGF-β1 siRNA lentivirus to kidney tissues). Rats with different expression levels of TGF-β1 were administered 1,25(OH)2D3 (0.03 μg/kg/d) or peanut oil as control. DN rats were treated only with peanut oil. All rats were randomly divided into five groups (n = 6 per group): TGF-β1 overexpression + oil, TGF-β1 overexpression + 1,25(OH)2D3, TGF-β1 siRNA + oil, TGF-β1 siRNA + 1,25(OH)2D3, and DN + oil groups. After 37 days, kidney samples were collected and the expression of TGF-β1 and miR-130b was determined by real-time PCR, western blotting, and immunohistochemistry. Hematoxylin and eosin staining and Masson staining were used to evaluate kidney morphological and fibrogenic changes. Differences were determined using ANOVA and Student's t-test. Results RT-PCR, western blotting, and immunohistochemistry revealed that interference of TGF-β1 significantly decreased mRNA and protein levels of TGF-β1 in renal tissues of DN rats compared to those in renal tissues of rats overexpressing TGF-β1 (p < 0.05). Histological analysis showed that upregulated TGF-β1 led to disorganized kidney structure and severe kidney fibrosis. The expression of miR-130b was significantly lowered upon lentivirus-mediated overexpression of TGF-β1 than upon downregulation of TGF-β1 (p < 0.05). Treatment with 1,25(OH)2D3 led to a significant reduction of TGF-β1 at the mRNA and protein levels (both p < 0.05), improvement of renal structure and fibrosis, and an increase in miR-130b expression (p < 0.05). Conclusion TGF-β1 can decrease the expression of miR-130b in kidney tissues of DN rats. Moreover, miR-130b may be involved in the protective effect of 1,25(OH)2D3 on renal fibrosis via TGF-β1.

Mesenchymal Stem Cells Reverse Diabetic Nephropathy Disease via Lipoxin A4 by Targeting Transforming Growth Factor β (TGF-β)/smad Pathway and Pro-Inflammatory Cytokines.

BackgroundDiabetic nephropathy (DN) is the leading cause of end-stage renal disease. Mesenchymal stem cells (MSCs) treatment has been proved to be effective in DN models by protecting renal function and preventing fibrosis. However, the underlying mechanism is unclear. Previous research indicated diabetes and associated complications may be attributed to failed resolution of inflammation, which is deliberately regulated by pro-resolving lipids, including lipoxins (LXs), resolvins (Rv) D and E series, protectins, and maresins. In this study, we monitored pro-resolving mediators in a DN model to explore the mechanism of MSCs treatment.Material/MethodsThe DN model was induced by STZ injection in SD rats. UPLC-MS/MS was performed to determine pro-resolving lipids in kidney tissue and serum of DN model before and after MSCs treatment, as well as in supernatants of HBZY-1-MSCs co-culture.ResultsLXA4 was highly accumulated in renal tissue of DN rats with MSCs treatment; ex vivo, LXA4 was significantly increased in the supernatants of HBZY-1 cells co-cultured with MSCs in a high-glucose (HG) medium. Western blot analysis indicated that ALX/FPR2, the receptor of LXA4, was markedly expressed in renal tissue of the DN-MSC group and HBZY-1 after incubating with MSCs in HG. Intraperitoneal injection of LXA4 inhibited renal fibrosis by targeting TGF-β/Smad signaling and downregulated serum TNF-α, IL-6, IL-8, and IFN-γ in DN rats. Notably, all the protective effects induced by MSCs or LXA4 were abolished by ALX/FRP2 blocking.ConclusionsOur results demonstrate that MSCs intervention prevented DN procession via the LXA4-ALX/FPR2 axis, which inhibited glomerulosclerosis and pro-inflammatory cytokines, eventually contributing to kidney homeostasis.

Upregulation of microRNA-424 relieved diabetic nephropathy by targeting Rictor through mTOR Complex2/Protein Kinase B signaling.

To investigate the role of miR-424 in diabetic nephropathy (DN) and its relationship with Rictor in mammalian target of rapamycin (mTOR) C2/Akt signaling. The western blot analysis and real-time polymerase chain reaction were used to determine the differential expression of Rictor, mTOR, and miR-424 in DN rats. The upregulation of miR-424 was achieved by caudal vein injection of miR-424 mimics. The renal lesion was evaluated by hematoxylin-eosin staining (H&E) and periodic acid schiff staining. The dual-luciferase reporter assay was conducted to determine the binding target of miR-424. The effect of miR-424 upregulation on apoptosis was detected by the terminal deoxynucleotidyl transferase-mediated 2-Deoxyuridine-5-Triphosphate (dUTP) nick-end labeling assay and western blot analysis. A significantly lower expression of miR-424 and a significantly higher expression of Rictor and mTOR were found in renal tissues of DN rats. The upregulation of miR-424 improved renal lesion and DN symptoms of blood glucose level, urine protein level, body weight, creatinine level, blood urea nitrogen, and KW/BW ratio. The upregulation of miR-424 could significantly reduce apoptosis rates of tissue cells by decreasing the expression levels of caspase-3 and Bax as well as increasing the level of Bcl-2. Furthermore, Rictor was the direct target for miR-424, and upregulation of miR-424 inhibited Rictor through Akt signaling in renal tissue of DN rats and high-glucose-treated human glomerular mesangial cells. miR-424 contributes to alleviating the symptoms in DN rat models by targeting Rictor through mTORC2/Akt signaling.

Combination of leflunomide and benazepril reduces renal injury of diabetic nephropathy rats and inhibits high-glucose induced cell apoptosis through regulation of NF-κB, TGF-β and TRPC6

Objective: To investigate effects of combination use of leflunomide and benazepril on diabetic nephropathy (DN) both in vivo and in vitro.Methods: The streptozotocin (STZ) induced Sprague-Dawley rats were treated with leflunomide (15 mg/kg/d), benazepril (15 mg/kg/d) or both the two drugs. Fasting blood glucose (FBG) and renal function indexes including blood urea nitrogen (BUN), serum creatinine (Scr), and proteinuria and kidney/body weight ratio (KW/BW) were measured. HE staining was used for histological analysis. The rat glomerular mesangial cells (RMCs) were treated with high-glucose (150 mg/ml) and the leflunomide and benazepril with both concentrations of 50 μmol/l were used to treat the high-glucose induced cells. TUNEL assay was used for measurement of cell apoptosis. Western blotting was conducted to determine expression of nuclear factor Kappa B (NF-κB), transforming growth factor-β (TGF-β) and transient receptor potential canonical 6 (TRPC6).Results: The body weight was significantly lower and all indexes of FBG, BUN, Scr, proteinuria and KW/BW ratio, GFR, as well as inflammatory factors TNF-α and IL-6 were significantly increased in the DN group after STZ treatment for 4 weeks. The treatment with leflunomide, benazepril or the both dramatically reduced the above effects induced by STZ, and the alteration was the most significant in the combination group. Treatment of leflunomide and benazepril significantly reduced expression levels of NF-κB, TGF-β and TRPC6 in renal tissues of DN rats as well as in high-glucose induced RMCs. It was also observed leflunomide and benazepril reduced high-glucose induced cell apoptosis of RMCs.Conclusion: The combination use of leflunomide and benazepril could improve the renal function and reduce the renal injury of DN rats and could reduce the levels of NF-κb, TGF-β and TRPC6 in both DN rats and high-glucose induced RMCs.