Renalase regulates renal tubular injury in diabetic nephropathy via the p38MAPK signaling pathway.

Diabetic nephropathy (DN) is an important complication of diabetes and the leading cause of end-stage renal disease globally. Renal tubular damage occurs to varying degrees in the early stages of DN prior to glomerular damage. Renalase (RNLS) is an amine oxidase, which is produced and secreted by the renal tubular epithelial cells. RNLS is reportedly closely related to renal tubular injury in acute and chronic kidney diseases. Herein, we aimed to evaluate the changes in tubular RNLS expression in DN and its correlation with DN-associated renal tubular injury. Conditional permanent renal tubular epithelial rat-cell line NRK-52E was transfected with pcDNA3-RNLS plasmid or administered recombinant rat RNLS protein and high glucose (HG) dose. A total of 22 adult Sprague-Dawley rats were randomly divided into the control (CON, n = 10) or diabetic nephrology (DN, n = 12) group. Random blood glucose levels of the rats were measured by sampling of the caudal vein weekly. After 8 weeks, the rat's body weight, 24-h urinary albumin concentration, and right kidney were evaluated. Our study suggested the decreased expression levels of RNLS in renal tissue and renal tubular epithelial cells in DN rats, accompanied by renal tubulointerstitial fibrosis, apoptosis of renal tubular epithelial cells, and activation of the p38MAPK signal pathway. Reversing the low RNLS expression can reduce the level of p38MAPK phosphorylation and delay renal tubular injury. Thus, the reduction of renal tubular RNLS expression in DN mediates tubulointerstitial fibrosis and cell apoptosis via the activation of the p38MAPK signal pathway. RNLS plays a key mediating role in DN-associated tubular injury via p38MAPK, which provides new therapeutic targets and a theoretical basis for early prevention and treatment of DN.

Hypoxia-inducible factor prolyl hydroxylase inhibitors for anaemia in maintenance dialysis: a meta-analysis.

BACKGROUND: Anaemia is a common complication of end-stage renal disease (ESRD) that relies on dialysis. Hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHI) is a new class of small-molecule oral drugs for the treatment of anaemia in chronic kidney disease. They demonstrate several advantages over traditional exogenous erythropoietin (EPO). We conducted a meta-analysis of studies that compared the efficacy of HIF-PHI in erythropoiesis and iron metabolism, and its safety with EPO in maintenance dialysis patients. METHODS: A sensitive search strategy in the PubMed, EMBASE and Cochrane databases identified all citations for randomised controlled trials (RCTs) comparing HIF-PHI agents with EPO/placebo through December 2021. RESULTS: Fourteen RCTs were identified, which included 2738 patients. No statistical difference was found in haemoglobin increase (p = 0.37) between HIF-PHI treatment and EPO using the random-effects model. HIF-PHI administration upregulated transferrin (MD 36.12, 95% CI 27.04-45.20) and soluble transferrin receptors (sTfR) (MD 1.28, 95% CI 0.44-2.13), but did not statistically reduce hepcidin level (p = 0.37). Total and LDL-cholestrol levels were suppressed by HIF-PHI (MD - 0.99, 95% CI - 1.34 to - 0.63) (MD - 0.99, 95% CI - 1.34 to - 0.64), while triglyceride (TG) was not different between HIF-PHI and EPO (p = 0.74). The total incident rates of treatment-emergent adverse events (TEAE) (p = 0.20) from HIF-PHI treatment were not different from those of erythropoietin, while the treatment-emergent serious adverse events (TSAE) (p = 0.02) were higher in the HIF-PHI group than those in the EPO controls with the fixed-effect model. CONCLUSION: HIF-PHI could effectively upregulate and maintain haemoglobin levels in patients with anaemia receiving maintenance dialysis. Furthermore, HIF-PHI could elevate iron metabolism activity and utility without inducing treatment-associated serious adverse events. Robust data from larger RCTs with longer treatment duration and follow-up are needed.

Association Between IL-6 Polymorphisms and Diabetic Nephropathy Risk: A Meta-analysis.

BACKGROUND: The objective of this work was to evaluate the relevance of frequent interleukin-6 (IL-6) polymorphisms and diabetic nephropathy (DN) susceptibility by a systematic meta-analysis. MATERIALS AND METHODS: The included studies related to the relationship between IL-6 and DN risk were searched from Pubmed, Embase and the Cochrane Library, and the Newcastle-Ottawa Scale was used to evaluate the study quality. A heterogeneity test was performed to determine the appropriate effect models based on the Q test and I2 statistic. Odds ratios with 95% confidence intervals were used to determine the strength of associations. Afterwards, subgroup analysis was conducted to assess the effect of specific factors on the corresponding results. Additionally, publication bias and sensitivity analysis were also undertaken. RESULTS: In total, 11 eligible articles were obtained. The meta-analysis revealed that the "C"allele of IL-6 rs1800795 was related to the decreased risk of DN (C versus G: P = 0.0471). The "G"allele of IL-6 rs1800796 was predominately associated with higher DN risks (GG versus CC: P = 0.0194; GG versus CC + GC: P = 0.0196). The "C"allele of IL-6 rs1800797 was implicated with higher prevalence of DN (C versus G: P = 0.0001; CC versus GG: P = 0.0003; CC versus GG + CG: P = 0.0227; CC + CG versus GG: P = 0.0001) while IL-6 rs2069837 and rs2069840 were not correlated with the susceptibility to DN. CONCLUSIONS: This meta-analysis indicated that IL-6 rs1800795, rs1800796 and rs1800797 played important roles in DN development while IL-6 rs2069837 and rs2069840 might not be related to DN.

Diagnostic value of neutrophil gelatinase-associated lipocalin in diabetic nephropathy: a meta-analysis.

Purpose: This meta-analysis aimed to determine the diagnostic performance of neutrophil gelatinase-associated lipocalin (NGAL) in diabetic nephropathy (DN). Methods: We searched the PubMed, Embase, Wanfang, and China National Knowledge Infrastructure databases for articles published up to 24 April 2019. The meta-analyses were conducted by Stata 11.0, and diagnostic accuracy, sensitivity, specificity, negative and positive likelihood ratios (NLR and PLR), diagnostic odds ratio (DOR), and receiver operating characteristic (ROC) curve data were pooled. Moreover, heterogeneity and small trials bias were evaluated. Results: Six cross-sectional studies were included in the meta-analysis. For the studies of microalbuminuria versus normoalbuminuria, the estimates (95% confidence interval) were as follows: sensitivity, 0.75 (0.51-0.89); specificity, 0.78 (0.70-0.84); PLR, 3.37 (2.49-4.56); NLR, 0.33 (0.16-0.69); DOR, 10.31 (4.05-26.25); and area under the ROC curve (AUC), 0.81 (0.77-0.84). For the studies of microalbuminuria + macroalbuminuria versus normoalbuminuria, the results were as follows: sensitivity, 0.83 (0.66-0.93); specificity, 0.88 (0.67-0.97); PLR, 7.20 (1.97-26.31); NLR, 0.19 (0.08-0.46); DOR, 37.83 (4.84-295.65); AUC, 0.92 (0.90-0.94). Deeks' funnel plot suggested that small trials bias was insignificant in this study. Conclusions: Our findings suggest that NGAL is a potential diagnostic marker for patients with DN and that its diagnostic value for microalbuminuria + macroalbuminuria is superior to that for microalbuminuria. Highlights The first meta-analysis to investigate NGAL diagnostic role in DN. NGAL is valuable for the early diagnosis of DN. The diagnostic value of NGAL in microalbuminuria + macroalbuminuria was much higher.

Expression, purification and biological effect of a novel single chain Fv antibody and protamine fusion protein for the targeted delivery of siRNAs to FGFR3 positive cancer cells

Background: Gain-of-function of fibroblast growth factor receptor 3 (FGFR3) is involved in the pathogenesis of many tumors. More and more studies have focused on the potential usage of therapeutic single-chain Fv (ScFv) antibodies against FGFR3. RNA interference (RNAi) has been considered as a promising therapeutic method against cancer. A tool which can deliver small interference RNAs (siRNAs) into FGFR3 positive cancer cells is very promising for anti-tumor therapy. Results: In this study, a novel fusion protein R3P, which consists of FGFR3-ScFv and protamine, was generated in Escherichia coli by inclusion body expression strategy and Ni-NTA chromatography. Its yield reached 10 mg per liter of bacterial culture and its purity was shown to be higher than 95%. 1 µg of R3P could efficiently bind to about 2.5 pmol siRNAs and deliver siRNAs into FGFR3 positive RT112 and K562 cells. Annexin V staining results showed that R3P can deliver the amplified breast cancer 1 (AIB1) siRNAs to induce RT112 cell apoptosis. Conclusion: These results indicated that R3P was a promising carrier tool to deliver siRNAs into FGFR3 positive cancer cells and to exert anti-tumor effect.

12-Lipoxygenase Inhibition on Microalbuminuria in Type-1 and Type-2 Diabetes Is Associated with Changes of Glomerular Angiotensin II Type 1 Receptor Related to Insulin Resistance.

(1) BACKGROUND: 12-lipoxygenase (12-LO) is involved in the development of diabetic nephropathy (DN). In the present study, we investigated whether 12-LO inhibition may ameliorate type-2 DN (T2DN) by interfering with insulin resistance (IR); (2) METHODS: Rat glomerular mesangial cells, glomeruli and skeletal muscles were isolated and used in this study. Kidney histological changes were confirmed by periodic-acid Schiff staining; mRNA expression was detected by competitive reverse transcription polymerase chain reaction; and the protein level was determined by Western blot and the enzyme-linked immunosorbent assay, respectively; (3) RESULTS: The inhibition of 12-LO attenuated microalbuminuria (MAU) increases in type-2 diabetic rats, but not in type-1 diabetic rats. Infusion of 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) significantly increased the expression of angiotensin II (Ang II) and Ang II type 1 receptor (AT1R), but decreased the expression of AT1R-associated protein (ATRAP) in rat glomeruli, compared to the control. An in vitro study revealed that both 12(S)-HETE and insulin upregulated AT1R expression in rat mesangial cells. In the presence of p38 mitogen-activated protein kinase (MAPK) inhibitor, SB202190, the 12(S)-HETE-induced ATRAP reduction was significantly abolished. Interestingly, 12-LO inhibition did not influence AT1R expression in type-1 diabetic rats, but significantly abolished the increased AT1R and Ang II expression in glomeruli of type-2 diabetic rats. Furthermore, the inhibition of 12-LO significantly corrected impaired insulin sensitivity and fast serum insulin level, as well as the p-AMP-activated protein kinase (AMPK) reduction in skeletal muscle of type-2 diabetic rats; (4) CONCLUSION: The inhibition of 12-LO potentially ameliorated MAU by preventing IR through the downregulation of glomerular AT1R expression in T2DN.

p16ink4a Expression Is Increased through 12-Lipoxygenase in High Glucose-Stimulated Glomerular Mesangial Cells and Type 2 Diabetic Glomeruli.

BACKGROUND/AIMS: Arachidonic acid-metabolizing enzyme, 12-lipoxygenase (12-LO), is involved in the glomerular hypertrophy of diabetic nephropathy (DN), in which cyclin-dependent kinase inhibitors (CKIs) play important roles. However, it is unclear whether 12-LO regulates the expression of the CKI p16(ink4a) in DN. METHODS: Primary glomerular mesangial cells (MCs) and glomeruli isolated from rats were used in this study. The rats were fed a high-fat diet and given low-dose streptozotocin to induce type 2 diabetes. The 12-LO product, 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE), was infused through an osmotic minipump. Enzyme-linked immunosorbent assay, Western blot, and morphometric analyses were performed. RESULTS: High glucose (HG) increased the p16(ink4a) protein expression in MCs, but this increase was prevented by the 12-LO inhibitor, cinnamyl-3,​4-dihydroxy-α-cynanocinnamate (CDC). The levels of p-p38MAPK and p16(ink4a) in MCs were significantly elevated after the 12(S)-HETE treatment, whereas the p38MAPK inhibitor SB203580 prevented these increases. Compared with levels in control MCs, marked increases in p38MAPK activation and p16(ink4a) expression were observed in MCs plated on collagen IV, while the CDC treatment prevented these changes. Subcutaneous injection of CDC did not affect glucose levels, but completely attenuated the diabetes-related increases in the 12(S)-HETE content, p16(ink4a) expression, p-p38MAPK levels, glomerular volume, and the kidney/body weight ratio. Compared with levels in controls, p16(ink4a) and p-p38MAPK in the glomeruli derived from 12(S)-HETE-treated rats were significantly higher. CONCLUSIONS: 12-LO-p38MAPK mediates the upregulation of p16(ink4a) in HG-stimulated MCs and type 2-diabetic glomeruli, and new therapies aimed at 12-LO inhibition may prove beneficial in ameliorating diabetes-induced glomerular hypertrophy.