Diabetic Kidney Disease Group Research Articles

The Correlation between Thyroid Hormone Levels and the Kidney Disease Progression Risk in Patients with Type 2 Diabetes.

ObjectiveWe investigated the relationship between thyroid hormones and the risk of diabetic kidney disease (DKD) progression.MethodsA total of 452 patients with type 2 diabetes were included, and a cross-sectional analysis was performed. Urine albumin/creatinine ratio (ACR) and estimated glomerular filtration rate (eGFR) were used to diagnose persistent albuminuria and stage chronic kidney disease, respectively. The Kidney Disease: Improving Global Outcomes (KDIGO) clinical practice guideline was used to describe the risk of DKD progression (low, moderate, and high or very high risks).ResultsThe DKD group had higher levels of thyroid-stimulating hormone (TSH) and lower levels of free triiodothyronine (FT3) and free thyroxine (FT4) than the non-DKD group. The prevalence of thyroid dysfunction in the DKD group was significantly higher than in the non-DKD group, especially the prevalence of subclinical hypothyroidism. FT3 levels decreased gradually with the deterioration of DKD. TSH levels increased with an increasing KDIGO category. FT3 and FT4 levels were negatively correlated with serum creatinine levels and ACR, and positively correlated with eGFR. Contrastingly, TSH was positively correlated with ACR, and negatively correlated with eGFR. After adjustment, an increase in FT3 levels significantly reduced the risk of DKD [odds ratio, OR (95% confidence interval, CI)=0.58 (0.42–0.79)] and DKD progression [ORs (95% CIs)=0.65 (0.45–0.93) for the moderate risk group and 0.50 (0.33–0.74) for the high or very high-risk group, using the low-risk group as a reference]. FT3 levels below 4.30 pmol/L in men and 3.99 pmol/L in women were the cut-off points for an increased risk of DKD progression.ConclusionLow FT3 level is an independent risk factor for DKD and DKD progression. FT3 ≤4.30 pmol/L in men and ≤3.99 pmol/L in women will greatly increase the risk of kidney disease progression in patients with type 2 diabetes.

Insulin Resistance in Early and Advanced Diabetic Kidney Disease

Background: Insulin resistance (IR) is commonly seen in diabetic kidney disease (DKD) and could contribute to the progression of renal disease and cardiovascular risk. In this study, we aim to measure homeostasis model assessment IR (HOMA-IR) in DKD and see the effect of advancing kidney disease on HOMA IR. Material and Methods: We recruited 120 subjects with type 2 diabetes mellitus and divided them into people without kidney disease (controls; n = 20), early DKD (n = 40), and advanced DKD (n = 60). Biochemical tests including fasting plasma glucose and fasting serum C-peptide were done in 120 subjects. IR was calculated by the HOMA model in 109 subjects. Data were presented as median (interquartile range [IQR]). Univariable and multivariable analysis was done. Results: Median of HOMA-IR in the control group was 2.0 (IQR: 1.5–2.8; n = 20), early DKD group was 2.3 (1.8–2.9; n = 37), and advanced DKD group was 3.67 (1.6–3.9; n = 52). P = 0.03 indicated a significant increase in the HOMA IR with advancing kidney disease. Conclusion: In patients with DKD, with advancing kidney disease, there was a significant increase in the HOMA IR, a marker of IR. IR is a modifiable metabolic risk factor, and if it is managed by novel therapeutic ways, it might improve clinical outcomes in DKD.

A Retrospective Analysis of Hematologic Parameters in Patients with Early Diabetic Kidney Disease.

ObjectiveTo retrospectively analyze the hematologic parameters in patients with early Diabetic Kidney Disease (DKD) to define potential biomarkers that can be used to predict early DKD.Methods134 diabetic patients without nephropathy and 49 patients with early DKD were enrolled for this study and the hematologic parameters were retrospectively analyzed. Paired comparison was conducted by T-test and the predicting value of any statistically different parameter was tested using the Receiver Operating Characteristic curve (ROC) analysis model.ResultsThe number of Neutrophil (N) was higher (P <0.001) while monocyte (M) was lower (P <0.01) in the early DKD group than that of DM group without nephropathy. In addition, neutrophil to lymphocyte ratio (NLR) was higher while platelet to lymphocyte ratio (PLR) was significantly lower in the early DKD group(P <0.001). Results from ROC curve analysis showed the sensitivity and specificity of PLR to predict early DKD were 83.7% and 82.6%, respectively.ConclusionsPLR may be a potential hematologic parameter that can be used to predict early DKD.

Fibrinogen to Albumin Ratio as an Independent Risk Factor for Type 2 Diabetic Kidney Disease.

PurposeDiabetic kidney disease (DKD) is an inflammatory disease. This study aimed to investigate the association of fibrinogen to albumin ratio (FAR) with DKD.Patients and MethodsA total of 1022 type 2 diabetes mellitus (T2DM) patients with DKD and 1203 T2DM patients without DKD were enrolled in this study. Laboratory values including blood cell count, hemoglobin A1c, biochemical parameters, and fibrinogen and albumin creatinine ratio were recorded. Patients were classified according to tertile of admission FAR. Clinical parameters were compared between groups. Logistic regression, linear regression, ROC analysis and spline regression were carried out.ResultsFAR in the DKD group was significantly higher than that in the non-DKD group. FAR had the highest odds ratio as an independent risk factor for the development of DKD and the highest area under ROC curve for predicting DKD compared with albumin (ALB) or fibrinogen (FIB) alone. Simple linear regression analyses revealed a significant and linear correlation of FAR with neutrophil and neutrophil-to-lymphocyte ratio. FAR was an independent risk factor for development of DKD. Spline regression showed that there was a significant linear association between DKD incidence and continuous FAR value when it exceeded 67.3mg/g.ConclusionFAR is a stronger independent predictor of DKD than FIB and ALB. FAR is an independent risk factor for DKD development when it exceeded 67.3mg/g. FAR might be one of novel diagnostic biomarkers to predict and prevent DKD progression. However, a prospective study to validate the prognostic model is still needed.

The change of gut microbiota-derived short-chain fatty acids in diabetic kidney disease.

BackgroundPrevious studies found the dysbiosis of intestinal microbiota in diabetic kidney disease (DKD), especially the decreased SCFA‐producing bacteria. We aimed to investigate the concentration of the stool and serum short‐chain fatty acids (SCFAs), gut microbiota‐derived metabolites, in individuals with DKD and reveal the correlations between SCFAs and renal function.MethodsA total of 30 participants with DKD, 30 participants with type 2 diabetes mellitus (DM), and 30 normal controls (NC) in HwaMei Hospital were recruited from 1/1/2018 to 12/31/2019. Participants with DKD were divided into low estimated glomerular filtration rate (eGFR)(eGFR<60ml/min, n=14) and high eGFR (eGFR≥60ml/min, n=16) subgroups. Stool and serum were measured for SCFAs with gas chromatograph‐mass spectrometry.ResultsThe DKD group showed markedly lower levels of fecal acetate, propionate, and butyrate versus NC (p<0.001, p<0.001, p=0.018, respectively) [1027.32(784.21–1357.90)]vs[2064.59(1561.82–2637.44)]μg/g,[929.53(493.65–1344.26)]vs[1684.57(1110.54–2324.69)]μg/g,[851.39(409.57–1611.65)] vs[1440.74(1004.15–2594.73)]μg/g, respectively, and the lowest fecal total SCFAs concentration among the groups. DKD group also had a lower serum caproate concentration than that with diabetes (p=0.020)[0.57(0.47–0.61)]vs[0.65(0.53–0.79)]μmol/L. In the univariate regression analysis, fecal and serum acetate correlated with eGFR (OR=1.013, p=0.072; OR=1.017, p=0.032). The correlation between serum total SCFAs and eGFR showed statistical significance (OR=1.019, p=0.024) unadjusted and a borderline significance (OR=1.024, p=0.063) when adjusted for Hb and LDL. The decrease in serum acetate and total SCFAs were found of borderline significant difference in both subgroups (p=0.055, p=0.050).ConclusionThis study provides evidence that in individuals with DKD, serum and fecal SCFAs levels (fecal level in particular) were lowered, and there was a negative correlation between SCFAs and renal function.

Retinal Neuropathy in IGT Stage of OLETF Rats: Another Characteristic Change of Diabetic Retinopathy.

Aims We investigated the changes of retinal structure in normal glucose tolerance (NGT), impaired glucose tolerance (IGT), diabetes mellitus (DM), and diabetic kidney disease (DKD) stages in Otsuka Long-Evans Tokushima Fatty (OLETF) rats. Methods We assigned OLETF rats to four groups based on their OGTT results and 24 h urinary microalbumin (24 h UMA) levels: NGT, IGT, DM, and DKD groups. We observed the structural and the corresponding pathological changes and quantified the expression of HIF-1α, iNOS, NF-κB, VEGF, ICAM-1, and occludin in the retina. Results Significant damage to the retinal structure, especially in retinal ganglion cells (RGCs), was observed in the IGT stage. The expression of HIF-1α, iNOS, NF-κB, VEGF, and ICAM-1 was significantly upregulated, while that of occludin was downregulated. Conclusion Significant retinal neuropathy occurs in the IGT stage. Inflammation and hypoxia may damage the blood retina barrier (BRB), leading to diabetic retinopathy.

Diabetic kidney disease: Are the reported associations with single-nucleotide polymorphisms disease-specific?

BACKGROUNDThe genetic backgrounds of diabetic kidney disease (DKD) and end-stage kidney disease (ESKD) have not been fully elucidated.AIMTo examine the individual and cumulative effects of single-nucleotide polymorphisms (SNPs) previously associated with DKD on the risk for ESKD of diabetic etiology and to determine if any associations observed were specific for DKD.METHODSFourteen SNPs were genotyped in hemodialyzed 136 patients with diabetic ESKD (DKD group) and 121 patients with non-diabetic ESKD (NDKD group). Patients were also re-classified on the basis of the primary cause of chronic kidney disease (CKD). The distribution of alleles was compared between diabetic and non-diabetic groups as well as between different sub-phenotypes. The weighted multilocus genetic risk score (GRS) was calculated to estimate the cumulative risk conferred by all SNPs. The GRS distribution was then compared between the DKD and NDKD groups as well as in the groups according to the primary cause of CKD.RESULTSOne SNP (rs841853; SLC2A1) showed a nominal association with DKD (P = 0.048; P > 0.05 after Bonferroni correction). The GRS was higher in the DKD group (0.615 ± 0.260) than in the NDKD group (0.590 ± 0.253), but the difference was not significant (P = 0.46). The analysis of associations between GRS and individual factors did not show any significant correlation. However, the GRS was significantly higher in patients with glomerular disease than in those with tubulointerstitial disease (P = 0.014) and in those with a combined group (tubulointerstitial, vascular, and cystic and congenital disease) (P = 0.018).CONCLUSIONOur results suggest that selected SNPs that were previously associated with DKD may not be specific for DKD and may confer risk for CKD of different etiology, particularly those affecting renal glomeruli.

Qing-Re-Xiao-Zheng Formula Modulates Gut Microbiota and Inhibits Inflammation in Mice With Diabetic Kidney Disease.

Evidence indicates that the metabolic inflammation induced by gut microbiota dysbiosis contributes to diabetic kidney disease. Prebiotic supplementations to prevent gut microbiota dysbiosis, inhibit inflammatory responses, and protect the renal function in DKD. Qing-Re-Xiao-Zheng formula (QRXZF) is a Traditional Chinese Medicine (TCM) formula that has been used for DKD treatment in China. Recently, there are growing studies show that regulation of gut microbiota is a potential therapeutic strategy for DKD as it is able to reduce metabolic inflammation associated with DKD. However, it is unknown whether QRXZF is effective for DKD by regulating of gut microbiota. In this study, we investigated the reno-protective effect of QRXZF by exploring its potential mechanism between gut microbiota and downstream inflammatory pathways mediated by gut-derived lipopolysaccharide (LPS) in the kidney. High-fat diet (HFD) and streptozotocin injection-induced DKD mice model was established to assess the QRXZF effect in vivo. Mice treated with QRXZF for 8 weeks had significantly lower levels of urinary albumin, serum cholesterol and triglycerides. The renal injuries observed through histological analysis were attenuated as well. Also, mice in the QRXZF group had higher levels of Zonula occludens protein-1 (ZO-1) expression, lower levels of serum fluorescein-isothiocyanate (FITC)-dextran and less-damaged colonic mucosa as compared to the DKD group, implying the benefit role for the gut barrier integrity. QRXZF treatment also reversed gut dysbiosis and reduced levels of gut-derived LPS. Notably, the expression of toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB), which are important inflammation pathways in DKD, were suppressed in the QRXZF groups. In conclusion, our results indicated that the reno-protective effects of QRXZF was probably associated with modulating gut microbiota and inhibiting inflammatory responses in the kidney.

High concentrations of triglycerides are associated with diabetic kidney disease in new-onset type 2 diabetes in China: Findings from the China Cardiometabolic Disease and Cancer Cohort (4C) Study.

AimsThe aims of this study were to evaluate the associations of metabolic abnormalities with incident diabetic kidney disease (DKD) and to explore whether dyslipidaemia, particularly high fasting triglyceride (TG), was associated with the development of DKD.MethodsIn total, 11 142 patients with new‐onset type 2 diabetes with baseline estimated glomerular filtration rates (eGFR) ≥60 mL/min/1.73 m2 were followed up during 2011‐2016. Incident DKD was defined as eGFR <60 mL/min/1.73 m2 at follow‐up. Multiple logistic regression analysis was conducted to explore the relationship of metabolic abnormalities at baseline and at follow‐up with risks of DKD. High TG was defined by TG ≥1.70 mmol/L. Low high‐density lipoprotein cholesterol (HDL‐c) was defined by HDL‐c <1.0 mmol/L for men or <1.3 mmol/L for women.ResultsParticipants who developed DKD had higher levels of waist circumference and systolic blood pressure, and lower levels of HDL‐c at both baseline and follow‐up visits. The DKD group also had higher levels of post‐load plasma glucose and TG at follow‐up. Multivariate logistic regression analysis revealed that both high TG at baseline [odds ratio (OR) = 1.37, p = .012) and high TG at follow‐up (OR = 1.71, p < .001) were significantly associated with increased risks of DKD. Patients with high TG levels at both baseline and follow‐up had higher risk of DKD compared with constantly normal TG (OR = 1.65, p < .001) after adjustment for covariates.ConclusionsIn a large population of patients with new‐onset type 2 diabetes, a high TG level was an independent risk factor for the development of DKD. Tight TG control might delay the occurrence of DKD.

Clinical Predictors of Nondiabetic Kidney Disease in Patients with Diabetes: A Single-Center Study

Background Although diabetic kidney disease (DKD) could affect up to one-third of patients with diabetes mellitus (DM), these patients can develop kidney diseases different from DKD, or these conditions can superimpose on DKD. Several potential predictors of nondiabetic kidney disease (NDKD) have been proposed, but there are no definitive indications available for kidney biopsy in diabetic patients. Methods We designed a single-center, cross-sectional, and retrospective cohort study to identify clinical and laboratory factors associated with a diagnosis of NDKD after native kidney biopsy in diabetic patients and to investigate differences in time to end-stage kidney disease (ESKD) in patients with a diagnosis of DKD and NDKD. Results Of 142 patients included in our analysis, 89 (62.68%) had a histopathological diagnosis of NDKD or mixed NDKD + DKD. Patients in the NDKD group had significantly lower HbA1C, lower prevalence of diabetic retinopathy (DR), and less severe proteinuria, and there was a lower proportion of patients with nephrotic syndrome; the DKD group had significantly lower proportion of patients with hematological conditions. In the multivariate binary logistic regression, only absence of DR and presence of a hematological condition significantly predicted NDKD after adjustment for age and sex. Time to ESKD was significantly higher in patients with NDKD or mixed forms than in those with DKD. Conclusions After a careful selection, more than half of kidney biopsies performed in diabetic patients can identify NDKD (alone or with concomitant DKD). Absence of DR and coexistence of a hematological condition (especially MGUS) were strong predictors of NDKD in our cohort.

Comparative study on therapeutic effects of human stem cells sin experimental diabetic nephropathy

Background Diabetic kidney disease (DKD) is the most common cause of end-stage renal disease. The pathophysiology of DKD includes oxidative stress, the formation of advanced glycation end-products, cellular signaling, and renewal processes, so they are potential targets for future drug therapy. One of these therapies is stem cells, as there is no specific treatment for diabetic nephropathy. Objective This study aimed to compare the therapeutic effects of human hematopoietic or mesenchymal stem cells in the treatment of experimentally induced diabetic nephropathy in rats. Materials and methods A total of 40 Swiss albino male rats were used in the present study. The animals were randomly divided into four groups: group I, control group (10 rats); group II, DKD group (10 rats), where streptozotocin-induced DKD rat models were used; group III, DKD group (10 rats), where streptozotocin-induced DKD rat models were used; group III, DKD treated with mesenchymal stem cells (MSCs); and group IV, DKD treated with hematopoietic endothelial progenitor stem cells (EPSCs). The four studied groups were assessed for renal function tests, serum CD44, interleukin 6, interleukin 18, intercellular adhesion molecule-1), neutrophil gelatinase-associated lipocalin, total antioxidant capacity, and serum rat malondialdehyde. Results There were significant differences between both DKD treated with MSCs and those treated with hematopoietic EPSCs groups and the DKD group without treatment, whereas there was no statistically significant difference between both stem cells-treated groups and the control group. Conclusion This study showed that hematopoietic EPSCs have the same renoprotective effect when compared with MSCs.

Phosphatidylserine on Blood Cells and Microparticles Contributes to the Hyper-Coagulable State in Diabetic Kidney Disease

Introduction: Patients with diabetes mellitus (DM) are considered to be prothrombotic with major disturbances in hemostasis that are associated with an increased risk of venous thromboembolism, especially when microvascular complications of diabetes mellitus occur, such as diabetic kidney disease (DKD). Additionally, microthrombosis in glomerular capillaries could decrease the kidney blood flow and induce renal insufficiency. However, patients with DKD do not receive routine antithrombotic therapy. Therefore, more study is needed on the mechanisms of coagulation in DKD with the aim of identifying therapeutic targets. Recent studies have shown that phosphatidylserine (PS), a membrane constituent, plays an important role in the thrombosis. However, relatively little is known about the precise role of PS in the prothrombotic state of DKD. Our objectives were to study the effect of increased PS exposure on microparticles (MPs) and the outer membrane of MP-origin cells in DKD patients, and to evaluate its effect on procoagulant activity (PCA).Methods: DKD patients (n = 90) were divided into three groups based on urinary albumin excretion rate (UAER), defined as normoalbuminuria (<30 mg/24 h), microalbuminuria (30-299 mg/24 h), or macroalbuminuria (>300 mg/24 h), and compared to healthy controls (n = 30). Lactadherin was used to quantify PS exposure on MPs and their original cells. Healthy blood cells and human umbilical vein endothelial cells (HUVECs) were treated with 25 mM, 5 mM or 2.5 mM glucose and then PS exposure was measured. PCA of MPs and cells was evaluated by clotting time and purified coagulation complex assays. Fibrin production was determined by turbidity. Phosphatidylserine exposure and fibrin strands were observed using confocal microscopy.Results: Usingflow cytometry, we found that PS+ MPs (derived from platelets, erythrocytes, HUVECs, neutrophils, monocytes and lymphocytes) and blood cells (BCs) were significantly higher in patients than in controls. Furthermore, the number of PS+ MPs and BCs in patients with macroalbuminuria was significantly higher than in patients with normoalbuminuria. Similarly, we observed markedly elevated PS exposure on HUVECs cultured with serum from patients with macro-DKD versus serum from patients with micro-DKD or normo-DKD. In addition, circulating PS+ MPs cooperated with PS+ cells, contributing to markedly shortened coagulation time and dramatically increased FXa/thrombin generation and fibrin formation in each DKD group. Confocal microscopy images demonstrated co-localization of fibrin with PS on ECs. Moreover, blockade of exposed PS on MPs and cells with lactadherin inhibited PCA by approximately 80%. Additionally, both hyperglycemia (25 mmol/L) and hypoglycemia (2.5 mmol/L) induced increased PS exposure on cells compared to normoglycemia (5 mmol/L). Linear regression analysis showed that levels of PS+ BCs and MPs were positively correlated with uric acid and proteinuria, but negatively correlated with GFR.Conclusions: Our results suggested that PS+ MPs and blood cells play a prominent role in inducing the hypercoagulable and prothrombotic state in patients with DKD. The occurrence of abnormal glucose during the hypoglycemic therapy is associated with hypercoagulation in DKD. PS+ cells and MPs likely support thrombosis in renal vascular and result in renal dysfunction and disease progression. Our results demonstrate that PS provides binding sites for FXa and prothrombinase complexes and promotes thrombin formation. Thus, directly targeting FXa and prothrombinase complexes might decrease thrombotic and renal disease risk. The results from this study suggest that future research should focus on discovery and deployment of direct PS inhibitors.Disclosure of Interest: Jialan Shi has a patent for the use of lactadherin as a PS probe. DisclosuresNo relevant conflicts of interest to declare.

Ginkgo Biloba Extract EGB761 Ameliorates the Extracellular Matrix Accumulation and Mesenchymal Transformation of Renal Tubules in Diabetic Kidney Disease by Inhibiting Endoplasmic Reticulum Stress.

The study is aimed at investigating the effects of Ginkgo biloba extract EGB761 on renal tubular damage and endoplasmic reticulum stress (ERS) in diabetic kidney disease (DKD). A total of 50 C57BL/6 N mice were randomly divided into the normal group, DKD group, DKD+EGB761 group (36 mg/kg), and DKD+4-phenylbutyrate (4-PBA) group (1 g/kg). The DKD model was replicated by high-fat diet combined with intraperitoneal injection of streptozotocin (STZ). Renal tubular epithelial cells (HK-2) were divided into the control group, high-glucose group (30 mmol/L), EGB761 group (40 mg/L, 20 mg/L, 10 mg/L), TM group, and TM+4-PBA group. After 8 weeks of administration, expressions of serum creatinine (Scr), blood urea nitrogen (BUN), 24 h urinary protein (24 h Pro), fasting blood glucose (FBG), β2-microglobulin (β2-MG), and retinol binding protein 4 (RBP4) of mice were tested. The pathological changes of renal tissue were observed. The expressions of extracellular matrix (ECM) accumulation and epithelial-mesenchymal transition (EMT) markers α-smooth muscle actin (α-SMA), E-cadherin, fibronectin, and collagen IV, as well as the ERS markers GRP78 and ATF6, were tested by Western blot, qPCR, immunohistochemistry, or immunofluorescence. EGB761 could decrease the Scr, BUN, 24 h Pro, and FBG levels in the DKD group, alleviate renal pathological injury, decrease urine β2-MG, RBP4 levels, and decrease the expression of α-SMA, collagen IV, fibronectin, and GRP78, as well as ATF6, while increase the expression of E-cadherin. These findings demonstrate that EGB761 can improve renal function, reduce tubular injury, and ameliorate ECM accumulation and EMT in DKD kidney tubules, and the mechanism may be related to the inhibition of ERS.

Evaluation of Neutrophil–lymphocyte Ratio and Platelet–lymphocyte Ratio as Markers of Diabetic Kidney Disease in Bangladeshi Patients with Type 2 Diabetes Mellitus

Background: The roles of neutrophil–lymphocyte ratio (NLR) and platelet–lymphocyte ratio (PLR) as novel surrogate markers of diabetic kidney disease (DKD) are promising. Objective: The aim of this study was to evaluate whether NLR and PLR can predict DKD in type 2 diabetes mellitus (T2DM). Subjects and Methods: This cross-sectional study was conducted from July 2018 to June 2019, among 312 patients with T2DM recruited from eight tertiary hospitals throughout Bangladesh. Complete blood count (CBC) was performed in fully automated hematology analyzers, and NLR and PLR were calculated. The urine albumin-to-creatinine ratio (ACR) was measured in a random single-voided urine sample. The diagnosis of DKD was made based on the presence of albuminuria (ACR ≥30 mg/g) and/or reduced estimated glomerular filtration rate (eGFR &lt;60mL/min/1.73 m2) in the absence of signs or symptoms of other primary causes of kidney damage. Results: Among 312 study subjects, 150 (48.1%) had DKD. The mean age (51.9 ± 11.9 vs. 48.6 ± 9.6 years), fasting plasma glucose (10.3 ± 4.1 vs. 9.2 ± 3.0 mmol/L), 2-h postprandial plasma glucose (14.6 ± 5.6 vs. 12.8 ± 4.5 mmol/L), glycated hemoglobin (HbA1c) (9.2 ± 2.1 vs. 8.4 ± 1.9%), and serum triglyceride (213.6 ± 109.9 vs. 185.5 ± 100.8) were higher (P ≤0.05 in each instance) in subjects with DKD in comparison to those without DKD. DKD group had higher mean absolute neutrophil count (6.0 ± 2.0 vs. 5.4 ± 1.9 ×109/L, P = 0.022), platelet count (310.4 ± 87.3 vs. 287.0 ± 78.7 ×109/L, P = 0.013), NLR (2.16 ± 1.1 vs. 1.92 ± 0.96, P = 0.040), and PLR (115.45 ± 57.07 vs. 101.02 ± 40.06, P = 0.010). The levels of hemoglobin, total leukocyte count, absolute lymphocyte count, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and red cell distribution width were similar in the two groups. Conclusion: In this study, we found higher NLR and PLR in subjects with DKD than those without DKD. NLR and PLR may be considered as cheap, readily available alternative markers of DKD in resource-poor settings.

Active Vitamin D&amp;lt;sub&amp;gt;3&amp;lt;/sub&amp;gt; Protects Against Diabetic Kidney Disease by Regulating the JNK Signaling Pathway in Rats

Diabetic kidney disease (DKD) is an inflammatory disease caused by metabolic disorder. As an important signaling pathway in the inflammatory response, the JNK signaling pathway plays an crucial role in kidney injury in DKD. Vitamin D3 can reduce the inflammatory reaction and delay or even reverse DKD progression. Unfortunately, the mechanism by which vitamin D3 regulates DKD pathogenesis is unclear. This research established a DKD rat model and vitamin D3 and irbesartan were used as interventions. Then, urine and blood biochemistry; and inflammatory cytokine (IL-1 and IL-6), phosphorylated JNK pathway protein (MEK-4 and JNK1/2/3) and downstream factor (AP-1 and ATF-2) expression were assessed. We found that the DKD group showed body weight and insulin secretion were significantly decreased; significantly increased FPG, HOMA-IR and blood lipids; and significantly increased 24-h urinary protein (UPro) compared with normal group. Additionally, the levels of IL-1 and IL-6 and phosphorylated JNK pathway proteins were significantly elevated. These changes were improved by vitamin D3, especially at a low dosage. These results suggest that active vitamin D3 protects against DKD in rats by reducing IL-6 and IL-1 release, downregulating the JNK inflammatory signaling pathway, and inhibiting downstream transcription factor AP-1- and ATF-2-mediated kidney damage. This research provides a new theoretical support for vitamin D3 treatment of diabetic nephropathy.

Implication of podocin promoter variant haplotype in south Indian diabetic kidney patients

Type 2 diabetes (T2D) is a global epidemic, 40% of T2D individuals are associated with diabetic kidney disease (DKD) a major microvascular complication leading to end stage renal disease (ESRD). We aimed to screen for three podocin (NPHS2) gene promoter variants -535ins/del (CTTTTTTT)2, rs1079291(C > T) and rs12406197 (G > T) in T2D and DKD patients of south India to explore their role in the propensity to DKD. The present study enrolled a total of 200 T2D patients comprising an equal number of T2D and DKD patients and performed genotyping using PCR followed by Sanger sequencing. Individual genotype analysis performed for three podocin promoter variants yielded different results. -535 ins/del (CTTTTTT)2 and rs1079291(C > T) gene variants showed lack of association. However, the GG vs. GT + TT genotype of rs12406197(G > T) exhibited protective effect (OR = 0.51; 95% CI = 0.28–0.92; p = 0.03). A genotype dependent variation was noted with respect to rs1079291 mutant genotype for serum albumin, serum creatinine, eGFR and rs12406197 risk genotypes for HbA1c, serum albumin and serum creatinine within the DKD group. Haplotype block 2TT found to be a risk haplotype towards DKD (OR = 2.43; 95% CI = 1.17–5.03; p = 0.017). Diabetic individuals carrying mutant allele and the elevated levels of kidney functioning markers are vulnerable to nephropathy. Further 2TT promoter haplotype appears to play a role in the genetic susceptibility to DKD in south Indian patients suggesting that the low production/depletion of podocin may hinder the regeneration and replacement of podocytes. As the sample size of the present study is limited in order to test our contention large studies are warranted.

Ongoing donor-transmitted diabetic kidney disease in kidney transplant recipients with fair sugar control: a single center retrospective study

BackgroundTransplantation with a diabetic donor kidney may have some benefits compared to remaining on the waitlist for selected patients. However, we found that some kidney transplant recipients have ongoing donor-transmitted diabetic kidney disease (DT-DKD) despite fair blood sugar control. This study aimed to survey the incidence and clinical pattern of DT-DKD in kidney transplant recipients.MethodsWe retrospectively reviewed the medical records of kidney transplantations in our hospital. We found 357 kidney transplantations from February 2006 to April 2018. Among these, 23 (6.4%) diabetic donor kidney transplantations were done in the study period.ResultsAmong the 23 recipients, 6 (26.1%) displayed biopsy-proven DKD. Recipients with biopsy-proven DKD had longer dialysis vintage, higher proteinuria amount, lower last estimated glomerular filtration rate (eGFR), and a more rapid decline in the eGFR. The median fasting blood sugar level in the biopsy-proven DKD group was unexpectedly lower than the non-DKD group. Most of the pre-implantation frozen sections in biopsy-proven DKD group showed diabetic lesions worse than diabetic nephropathy (DN) class IIa. In the biopsy-proven DKD group, 5 recipients had no history of diabetes before or after transplantation. Among the 23 recipients, 5 (21.7%) were diagnosed with DT-DKD. Serial post-transplant biopsies showed the histological progression of allograft DN.ConclusionsTo the best of our knowledge, this is the first study to report the phenomenon of ongoing DT-DKD in kidney transplant recipients with fair blood sugar control. The zero-time pre-transplant kidney biopsy may be an important examination before the allocation of diabetic donor kidneys. Further study is needed to elucidate the possible mechanism of ongoing DT-DKD in non-diabetic recipients with fair blood sugar control as well as the impaction of pre-implantation diabetic lesion on the graft outcome.

Urinary miR-3137 and miR-4270 as potential biomarkers for diabetic kidney disease.

BackgroundAs one of the most prevalent diagnostic indicators of diabetic kidney disease (DKD), albumin‐to‐creatinine ratio (ACR) shows considerably limited predictive power in clinical application. We analyzed microarray expression profiling of urine to seek for differentially expressed miRNAs for potential biomarkers of DKD.MethodsUrine samples from type 2 diabetes mellitus (T2DM) patients with (30 mg/g < ACR < 300 mg/g, DKD group) or without DKD (ACR < 30 mg/g, DM group) were collected for miRNA microarray analysis. The differentially expressed miRNAs were screened by bioinformatics analysis and validated by quantitative real‐time PCR. Target genes of differentially expressed miRNAs were predicted in miRDB, Targetscan, and microRNA.org databases. We also conducted the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways analysis to explore for potential mechanisms in DKD.ResultsNine miRNAs were down‐regulated and seventeen miRNAs were up‐regulated in DKD group, compared to DM group. The levels of miR‐3137 and miR‐4270 in DKD group were 0.670 ± 0.505 and 2.116 ± 1.762 times than those in DM group, respectively, showing great significance. A total of 1076 target genes were simultaneously predicted by miRDB, Targetscan, and microRNA.org databases. According to the GO analysis results, disorders of endomembrane system may be one of the major pathological changes in DKD. In addition, Rap 1 signaling pathway is also altered obviously in DKD, discovered by the KEGG analysis.ConclusionMiR‐3137 and miR‐4270 show the potential for urinary biomarkers of DKD. The pathological changes of DKD may be related to disorders of endomembrane system and alternation of Rap1 signaling pathway.

Circulating Expression Level of LncRNA Malat1 in Diabetic Kidney Disease Patients and Its Clinical Significance

Background Long noncoding RNA MALAT1 is closely related to diabetes and kidney diseases and is expected to be a new target for the diagnosis and treatment of diabetic nephropathy. Objective This study aimed to explore the circulating expression level and significance of lncRNA Malat1 in patients with type 2 diabetes mellitus (T2DM) and diabetic kidney disease (DKD). Methods Quantitative real-time PCR (qPCR) was conducted to assess the expression of lncRNA Malat1 in 20 T2DM patients, 27 DKD patients, and 14 healthy controls, and then, the clinical significance was analyzed. Results LncRNA MALAT1 expression in peripheral blood mononuclear cells (PBMC) was significantly upregulated in T2DM and DKD groups when compared to control. Pearson's correlation analysis showed correlation of lncRNA MALAT1 levels with ACR, urine β2-microglobulin (β2-MG), urine α1-microglobulin (α1-MG), creatinine (Cr), and glycosylated hemoglobin (HbA1c), while negative with superoxide dismutase (SOD) (r = −0.388, P < 0.05). Binary regression analysis showed that ACR, creatinine, α1-MG, and LncRNA Malat1 were the risk factors for diabetic nephropathy with OR value of 1.166, 1.031, 1.031, and 2.019 (P < 0.05). The area under ROC curve (AUC) of DKD identified by the above indicators was 0.914, 0.643, 0.807, and 0.797, respectively. The AUC of Joint prediction probability of DKD recognition was 0.914, and the sensitivity and specificity of DKD diagnosis were 1.0 and 0.806, respectively. (Take ≥0.251 as the diagnostic cutoff point). Conclusion LncRNA Malat1 is highly expressed in DKD patients, and the combined detection of ACR, creatinine, α1-MG, and LncRNA Malat1 with diabetes mellitus may be the best way to diagnose diabetic nephropathy.

SO004IDENTIFICATION OF HUB GENES, FOCUSED ON CHEMOKINES AND CHEMOKINE RECEPTORS, ASSOCIATED WITH THE DECLINE OF RENAL FUCTION OF DIABETIC KIDNEY DISEASE BY WEIGHTED GENE CO-EXPRESSION NETWORK ANALYSIS

Abstract Background and Aims The fact that activation of the innate immune system and chronic inflammation are closely involved in the pathogenesis of diabetic Kidney disease (DKD). Recent studies have suggested the inflammatory process plays a crucial role in the progression of DKD. Identifying novel inflammatory molecules closely related to the decline of renal function is of significance in diagnosing and predicting the progression of DKD. The weighted gene co-expression network analysis (WGCNA) algorithm represents a novel systems biology method that provide the approach of association between gene modules and clinical traits to find the genes involvement into the certain phenotypic trait. The goal of this study was to identify hub genes and their roles in DKD from the gene sets associated with the decline of renal function by WGCNA. Method The Gene Expression Omnibus (GEO) database and “Nephroseq” website were searched and transcriptome study from DN biopsies with well-established clinical phenotypic data were selected for analysis. Next, we constructed a weighted gene co-expression network and identified modules negatively correlated with eGFR by WGCNA in the data of glomerular tissue. Functional annotations of the genes in modules negatively correlated with eGFR were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Through protein-protein interaction (PPI) analysis and hub gene screening, the hub genes were obtained. Furthermore, we compared the expression level of hub genes between DKD and normal control and drew ROC curves to determine the diagnosis value to DKD of these genes. Results The microarray-based expression datasets GSE30528 were screened out for analysis, which included glomeruli tissue of 9 cases of DKD and 13 cases of control. This microarray platform represented the transcriptome profile of 12411 well-characterized genes. Using WGCNA, a total of 19 gene modules were identified. Then module eigengene were analyzed for correlation with clinical traits of age, sex, ethnicity and eGFR and the “MEhoneydew1” module showed negative associated with eGFR (r=-0.58). GO functional annotation showed that these 551 genes in the “MEhoneydew1” module mainly enriched in the T cell activation. KEGG annotation showed mainly enriched in chemokine signaling pathway. Except for C3, top 10 hub genes, CCR5, CXCR4, CCR7, CCL5, CXCL8, CCR2, CCR1, CX3CR1, C3AR1 and C3, are all members of chemokines or chemokine receptors. Furthermore, we compared the expression level of these 9 genes between DKD and control, and found that all of these 9 genes increased in the DKD group, and the differences of 6 genes, CCR5, CCR7, CCL5, CCR2, CCR1, C3AR1, were of statistical significance. Linear correlation analysis showed that the expression of these 6 genes was negatively correlated with eGFR, and the ROC curve showed that the area under the curve could reach 0.812∼1.0. Conclusion We identified a panel of 6 hub genes focused on chemokines and chemokine receptors critical for decline of renal function of DKD using WGCNA. These genes may serve as biomarkers for diagnosis/prognosis and as putative novel therapeutic targets for DKD.