Nephropathy 1 Formula alleviates kidney injury by ameliorating mitochondrial dysfunction and pyroptosis in diabetic nephropathy through the TMAO-mROS-NLRP3 axis.

Nox4 is the major isoform of NADPH oxidase found in the kidney and contributes to the pathogenesis of diabetic nephropathy. However, the molecular mechanisms of increased Nox4 expression induced by hyperglycaemia remain to be elucidated. Here, the role of the connexin32-Nox4 signalling axis in diabetic nephropathy and its related mechanisms were investigated. Diabetes was induced in mice by low-dose streptozotocin (STZ) combined with a high-fat diet. Effects of connexin32 on Nox4 expression and on renal function and fibrosis in STZ-induced diabetic mice were investigated using adenovirus-overexpressing connexin32 and connexin32-deficient mice. Interactions between connexin32 and Nox4 were analysed by co-immunoprecipitation and immunofluorescence assays. Connexin32 was down-regulated in the kidneys of STZ-induced diabetic mice. Overexpression of connexin32 reduced expression of Nox4 and improved renal function and fibrosis in diabetic mice, whereas connexin32 deficiency had opposite effects. Down-regulation of fibronectin expression by connexin32 was not dependent on gap junctional intercellular communication involving connexin32. Connexin32 interacted with Nox4 and reduced the generation of hydrogen peroxide, leading to the down-regulation of fibronectin expression. Mechanistically, connexin32 decreased Nox4 expression by promoting its K48-linked polyubiquitination. Interestingly, Smurf1 overexpression inhibited K48-linked polyubiquitination of Nox4. Furthermore, connexin32 interacted with Smurf1 and inhibited its expression. Connexin32 ameliorated renal fibrosis in diabetic mice by promoting K48-linked Nox4 polyubiquitination and degradation via inhibition of Smurf1 expression. Targeting the connexin32-Nox4 signalling axis may contribute to the development of novel treatments for diabetic nephropathy.

Objective: To observe the protective effects of exogenous spermine on renal fibrosis induced by diabetic nephropathy (DN) and to explore its mechanism.Methods: Twenty-four male C57 mice were randomly divided into control group, type 1 diabetes group (TID) and spermine pretreatment group (TID+Sp, n=8 in each group). TID mice were induced by STZ (60 mg/kg), and TID+Sp mice were pretreated with spermine (5 mg/(kg·d)) for 2 weeks before STZ injection. The mice were killed at the 12th week. The renal function was determined by serum creatinine and urea nitrogen. HE, PAS and Masson staining were used to evaluate renal tissue injury and fibrosis. The expressions of matrix metalloproteinase (MMP-2, MMP-9) and collagen IV (Coll-IV) in the kidney of mice were detected by Western blot. Results: Compared with the control group, the blood glucose (5.67±0.22 vs 28.40±0.57 mmol/L), creatinine (14.33±1.22 vs 30.67±4.73 μmol/L) and urea nitrogen (6.93±4.94 vs 22.00±1.04 mmol/L) in the T1D group were increased significantly (P＜0.05), the glomerular basement membrane was thickened, the collagen was significantly increased, the expressions of MMP-2, MMP-9 and Coll-IV protein were increased (0.57±0.07 vs 1.06±0.20, 47.00±0.04 vs 1.29±0.09 and 0.42±0.16 vs 0.95±0.18,P＜0.05). Exogenous spermine significantly alleviates the above-mentioned changes. Conclusion: Exogenous spermine pretreatment could significantly alleviate renal fibrosis in diabetic mice by regulating the balance between MMPs and collagen.

The NLR family pyrin domain containing 3 (NLRP3) inflammasome is one of the best-characterized inflammasomes in humans and other mammals. However, knowledge about the NLRP3 inflammasome in nonmammalian species remains limited. Here, we report the molecular and functional identification of an NLRP3 homolog (DrNLRP3) in a zebrafish (Danio rerio) model. We found that DrNLRP3's overall structural architecture was shared with mammalian NLRP3s. It initiates a classical inflammasome assembly for zebrafish inflammatory caspase (DrCaspase-A/-B) activation and interleukin 1β (DrIL-1β) maturation in an apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC)-dependent manner, in which DrNLRP3 organizes DrASC into a filament that recruits DrCaspase-A/-B by homotypic pyrin domain (PYD)–PYD interactions. DrCaspase-A/-B activation in the DrNLRP3 inflammasome occurred in two steps, with DrCaspase-A being activated first and DrCaspase-B second. DrNLRP3 also directly activated full-length DrCaspase-B and elicited cell pyroptosis in a gasdermin E (GSDME)-dependent but ASC-independent manner. These two events were tightly coordinated by DrNLRP3 to ensure efficient IL-1β secretion for the initiation of host innate immunity. By knocking down DrNLRP3 in zebrafish embryos and generating a DrASC-knockout (DrASC−/−) fish clone, we characterized the function of the DrNLRP3 inflammasome in anti-bacterial immunity in vivo. The results of our study disclosed the origin of the NLRP3 inflammasome in teleost fish, providing a cross-species understanding of the evolutionary history of inflammasomes. Our findings also indicate that the NLRP3 inflammasome may coordinate inflammatory cytokine processing and secretion through a GSDME-mediated pyroptotic pathway, uncovering a previously unrecognized regulatory function of NLRP3 in both inflammation and cell pyroptosis. The NLR family pyrin domain containing 3 (NLRP3) inflammasome is one of the best-characterized inflammasomes in humans and other mammals. However, knowledge about the NLRP3 inflammasome in nonmammalian species remains limited. Here, we report the molecular and functional identification of an NLRP3 homolog (DrNLRP3) in a zebrafish (Danio rerio) model. We found that DrNLRP3's overall structural architecture was shared with mammalian NLRP3s. It initiates a classical inflammasome assembly for zebrafish inflammatory caspase (DrCaspase-A/-B) activation and interleukin 1β (DrIL-1β) maturation in an apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC)-dependent manner, in which DrNLRP3 organizes DrASC into a filament that recruits DrCaspase-A/-B by homotypic pyrin domain (PYD)–PYD interactions. DrCaspase-A/-B activation in the DrNLRP3 inflammasome occurred in two steps, with DrCaspase-A being activated first and DrCaspase-B second. DrNLRP3 also directly activated full-length DrCaspase-B and elicited cell pyroptosis in a gasdermin E (GSDME)-dependent but ASC-independent manner. These two events were tightly coordinated by DrNLRP3 to ensure efficient IL-1β secretion for the initiation of host innate immunity. By knocking down DrNLRP3 in zebrafish embryos and generating a DrASC-knockout (DrASC−/−) fish clone, we characterized the function of the DrNLRP3 inflammasome in anti-bacterial immunity in vivo. The results of our study disclosed the origin of the NLRP3 inflammasome in teleost fish, providing a cross-species understanding of the evolutionary history of inflammasomes. Our findings also indicate that the NLRP3 inflammasome may coordinate inflammatory cytokine processing and secretion through a GSDME-mediated pyroptotic pathway, uncovering a previously unrecognized regulatory function of NLRP3 in both inflammation and cell pyroptosis.

Neuroinflammation is widely recognized as a key factor in the pathogenesis of Alzheimer's disease (AD), alongside ß-amyloid deposition and the formation of neurofibrillary tangles. The NLR family pyrin domain containing 3 (NLRP3) inflammasome, part of the innate immune system, has been implicated in the neuropathology of both preclinical amyloid and tau transgenic models. Activation of the NLRP3 pathway involves an initial priming step, which increases the expression of Nlrp3 and interleukin (IL)-1β, followed by the assembly of the NLRP3 inflammasome complex, comprising NLRP3, ASC, and caspase-1. This assembly leads to the proteolytic maturation of the pro-inflammatory cytokines IL-1β and IL-18. Additionally, the NLRP3 inflammasome induces Gasdermin D (GSDMD) cleavage, forming membrane pores through which IL-1β and IL-18 are secreted. Inhibition of NLRP3 has been shown to enhance plaque clearance by modulating microglial activation. Furthermore, blocking NLRP3 in tau transgenic mice has been found to reduce tau phosphorylation by affecting the activity of certain tau kinases and phosphatases. In this study, organotypic brain slice cultures from P301S transgenic mice were treated with lipopolysaccharide (LPS) plus nigericin as a positive control or exposed to tau seeds (K18) to evaluate NLRP3 inflammasome activation. The effect of tau seeding on NLRP3 activity was further examined using Meso Scale Discovery (MSD) assays to measure IL1β secretion levels in the presence and absence of NLRP3 inhibitors. The role of NLRP3 activity was investigated in full-body Nlrp3 knockout mice crossbred with the tau transgenic P301S model. Additionally, full-body and microglia-selective Gsdmd knockout mice were crossbred with P301S mice, and tau pathology and neurodegeneration were evaluated at early and late stages of the disease using immunohistochemistry and biochemical assays. Activation of the NLRP3 pathway was observed in the mouse organotypic slice culture (OSC) model following stimulation with LPS and nigericin or exposure to tau seeds. However, Nlrp3 deficiency did not mitigate tauopathy or neurodegeneration in P301S mice in vivo, showing only a minor effect on plasma neurofilament (NF-L) levels. Consistently, Gsdmd deficiency did not alter tau pathology in P301S mice. Furthermore, neither full-body nor microglia-selective Gsdmd deletion had an impact on neuronal pathology or the release of pro-inflammatory cytokines. The absence of key components of the NLRP3 inflammasome pathway did not yield a beneficial effect on tau pathology or neurodegeneration in the preclinical Tau-P301S mouse model of AD. Nonetheless, organotypic slice cultures could serve as a valuable ex vivo mechanistic model for evaluating NLRP3 pathway activation and pharmacological inhibitors.

Acetaminophen (APAP) overdose is the primary cause of drug-induced acute liver failure in numerous Western countries. NLR family pyrin domain containing 3 (NLRP3) inflammasome activation serves a pivotal role in the pathogenesis of various forms of acute liver injury. However, the cellular source for NLRP3 induction and its involvement during APAP-induced hepatotoxicity have not been thoroughly investigated. In the present study, hematoxylin and eosin staining was performed to assess histopathological changes of liver tissue. Immunohistochemistry staining(NLRP3, Caspase-1, IL-1β, GSDMD and Caspase-3), western blotting (NLRP3, Caspase-1, IL-1β, GSDMD and Caspase-3) and RT-qPCR (NLRP3, Caspase-1 and IL-1β) were performed to assess the expression of NLRP3/GSDMD signaling pathway. TUNEL staining was performed to assess apoptosis of liver tissue. The serum expression levels of inflammatory factors (IL-6, IL-18, IL-1β and TNF-α) were assessed using ELISA and inflammation of liver tissue was assessed using immunohistochemistry (Ly6G and CD68) and RT-qPCR (TNF-α, Il-6, Mcp-1, Cxcl-1, Cxcl-2). A Cell Counting Kit-8 was performed to assess cell viability and apoptosis. Protein and gene expression were analyzed by western blotting (PCNA, CCND1) and RT-qPCR (CyclinA2, CyclinD1 and CyclinE1). Through investigation of an APAP-induced acute liver injury model (AILI), the present study demonstrated that APAP overdose induced activation of NLRP3 and cleavage of gasdermin D (GSDMD) in hepatocytes, both in vivo and in vitro. Additionally, mice with hepatocyte-specific knockout of Nlrp3 exhibited reduced liver injury and lower mortality following APAP intervention, accompanied by decreased infiltration of inflammatory cells and attenuated inflammatory response. Furthermore, pharmacological blockade of NLRP3/GSDMD signaling using MCC950 or disulfiram significantly ameliorated liver injury and reduced hepatocyte death. Notably, hepatocyte Nlrp3 deficiency promoted liver recovery by enhancing hepatocyte proliferation. Collectively, the present study demonstrated that inhibition of the NLRP3 inflammasome protects against APAP-induced acute liver injury by reducing hepatocyte pyroptosis and suggests that targeting NLRP3 may hold therapeutic potential for treating AILI.

Diabetic nephropathy (DN) is the leading cause of end-stage renal disease. We aimed to explore the role of RNA binding motif protein 15 (RBM15) in high glucose (HG)-induced pyroptosis of renal tubular epithelial cell, thus providing theoretical knowledge and new targets for DN treatment. HG-induced HK-2 cells were used to establish DN cell models. RBM15 expression was inhibited in HK-2 and detected. Cell viability was detected by cell counting kit-8. The levels of NLR family pyrin domain containing 3 (NLRP3), NLR family CARD domain containing 4 (NLRC4), gasdermin D (GSDMD)-N, and cleaved Caspase-1 were detected by Western blot assay. The levels of IL-1β and IL-18 were detected by enzyme linked immunosorbent assay. The enrichment of insulin-like growth factor 2 mRNA binding protein (IGF2BP2) and N6-methyladenosine (m6A) on NLRP3 and NLRC4 were analyzed by methylated RNA immunoprecipitation (MeRIP) and RIP. The stability of NLRP3 and NLRC4 mRNA was analyzed. The mechanism was verified by interfering NLRP3 and NLRC4 expression. RBM15 was highly expressed in HG-induced HK-2 cells. Inhibition of RBM15 reversed cell viability, inhibited inflammation, and alleviated pyroptosis. RBM15 promoted the expression of inflammasomes NLRP3 and NLRC4 through IGF2BP2-dependent m6A modification. RBM15 promoted the expression of inflammasomes NLRP3 and NLRC4 through IGF2BP2-dependent m6A modification, thereby promoting pyroptosis.

ObjectiveThe prevalence of type 2 diabetes mellitus (T2DM) and bone metabolism disorders increase with age. Diabetic kidney disease (DKD) is one of the most serious microvascular complications of T2DM, and bone metabolism disorders are closely linked to the occurrence of DKD. The relationship between bone turnover markers(BTMs) and the kidney disease in elderly patients with T2DM remains unclear. Therefore, this study aims to investigate the association between common BTMs and DKD in a large sample of elderly patients. The goal is to provide a basis for early identification of high-risk individuals for DKD among elderly T2DM patients from a bone metabolism perspective.MethodsIn this cross-sectional study, BTMs were collected from a cohort of 2,051 hospitalized Chinese patients. The relationships between 25-hydroxyvitamin D (25-OH-D), β-CrossLaps (β-CTX), osteocalcin (OSTEOC), intact parathyroid hormone (iPTH), and total type I collagen N-terminal propeptide (TP1NP), and DKD, as well as urinary albumin-to-creatinine ratio (UACR) and estimated glomerular filtration rate (eGFR) were analyzed using regression analysis and restrictive cubic spline (RCS) curves.ResultsHigher 25-OH-D levels were independently linked to a lower incidence of DKD and decreased UACR. The RCS curves showed a linear association of 25-OH-D and DKD, approaching the L-shape. β-CTX was independently and positively correlated with UACR. There is an independent positive correlation between OSTEOC and UACR and a negative correlation with eGFR. iPTH is independently and positively correlated with DKD incidence and UACR, and negatively correlated with eGFR. Additionally, the RCS curves showed a non-linear association of OSTEOC and iPTH and DKD, approaching the J-shape, and the point of inflection is 10.875 ng/L and 34.15 pg/mL respectively. There is an independent positive correlation between TP1NP and UACR incidence, and a negative correlation with eGFR. Risk estimates significantly increase with higher TP1NP levels in the RCS model.ConclusionBTMs are closely associated with kidney disease in elderly patients with T2DM. These discoveries potentially assist clinicians in establishing more preventive measures and targeted treatment strategies for elderly patients with T2DM.

Expression levels of serum vasohibin-1 and other biomarkers in type 2 diabetes mellitus patients with different urinary albumin to creatinine ratios

Background and Aims Correct identification of diabetic kidney disease (DKD) in type 2 diabetes mellitus (T2DM) patients is crucial to implement therapeutic interventions to prevent disease progression and premature death. Method We have analyzed the prevalence of DKD according to lab criteria and the rate of identification of DKD and/or chronic kidney disease (CKD) on 516,578 hospital care electronic medical records (EMR) in a tertiary hospital-based population using Savana Manager®. Results Out of 24,129 T2DM patients, 15,304 met inclusion criteria. DKD was defined as eGFR<60 ml/min/1.73m2 or urinary albumin to creatinine ratio (UACR) >30 mg/g or urinary protein to creatinine ratio (UPCR) >0.3 g/g after excluding acute kidney injury (AKI). A total of 4,526 (29.6%) T2DM patients had DKD according to lab criteria. However, the terms “CKD” or “DKD” were only present in 33.1% and 7.5% of the 4,526 EMR, with a hidden prevalence of CKD and DKD of 66.9% and 92.5%, respectively. Less severe kidney disease (lower UACR or UPCR, higher eGFR values), female sex, and lack of insulin prescription were associated with the absence of “DKD” or “CKD” terms in EMR (p<0.001) in the patients fulfilling laboratory criteria for DKD. However, while younger age (<70 years) was associated with a missing CKD diagnosis, older age (≥70 years) was associated with a missing DKD diagnosis in EMR. Conclusion In conclusion, the prevalence of DKD among T2DM patients according to laboratory data is higher than prevalence based on specific diagnosis written in EMR. This could imply underdiagnosis of DKD, especially in patients with less severe disease who may benefit the most from optimized therapy

Objective To investigate the relationship between platelet to lymphocyte ratio (PLR) and diabetic nephropathy (DN) in patients with type 2 diabetes mellitus (T2DM). Methods Eighty inpatients with T2DM treated in our hospital from July 2015 to December 2015 were included, and were divided into DN group and non-DN group (normo-albuminuria group) based on the guideline of prevention and management T2DM in China (2013 edition). The patients in DN group were assigned into micro-albuminuria and macro-albuminuria group according to the levels of urine albumin to creatinine ratio (UACR). Clinical and biochemical data were collected, and stasistical analysis were performed. Results Patients in DN group had higher PLR levels as compared to those in non-DN group (127.04±60.67 vs 92.23±36.44, P=0.003), also PLR discriminated among normo-, micro- and macro-albuminuria groups(92.23±36.44 vs 127.87±66.53 vs 125.57±50.83, P=0.010). However, there were no differences among the 3 groups, although PLR positively correlated to UACR(r=0.225, P=0.045). In binary regression analysis, PLR(OR=1.022, 95%CI: 1.004-1.041, P=0.017)and duration of diabetes(OR=1.037, 95%CI: 1.002-1.073, P=0.039)were predicted to be independent risk factors of DN. Conclusions PLR significantly elevated in T2DM patients with DN, correlated with UACR, and might be one of the independent risk factors for DN. Key words: Platelet count; Lymphocyte count; Type 2 diabetes; Diabetic nephropathy; Albuminuria

ObjectivesAmong all the diabetes complications brought on by persistent inflammation is diabetic kidney disease (DKD). One essential method of the inflammatory response's programmed cell death is anthrax. One of the main causes of diabetic renal disease progression in a high-glycemic environment is the lysis of renal resident cells.MethodThis investigation sought to determine whether Astragaloside IV (AS-IV)'s anti-pyroptosis action provides a protective function for the kidneys. For 12 weeks, db/db mice received 40 mg/kg of AS-IV by transgastric gavage. To validate the possible in vitro mechanism, mouse podocytes were cultivated for additional experiments.ResultsIn vitro, AS-IV led to a significant reduction in blood urea nitrogen (BUN), urine albumen-to-creatinine ratio (UACR), serum creatinine (CREA), and hyperglycemia in db/db mice and lessen the pathological alterations in the kidney. Moreover, pyrin structural domain of the NLR family pyrin domain containing 3 (NLRP3), cleaved-caspase-1, gasdermin D (GSDMD), IL-18, and IL-1β were down-expressed and podocyte markers podocin and nphs1 were up-regulated following AS-IV intervention. By silencing GSDMD, we demonstrated in vitro that HG-stimulated podocytes undergo pyroptosis. We also discovered that AS-IV can mitigate this pyroptosis. To confirm that AS-IV prevented the NLRP3 inflammasome from activating, the NLRP3 inhibitor CY-09 was employed. It was also discovered that AS-IV prevents the expression of TXNIP and NLRP3 as well as their interaction. GSDMD expression was significantly downregulated following TXNIP-siRNA treatment, whereas GSDMD expression was upregulated in TXNIP overexpression cells; this upregulation could be undone with AS-IV.ConclusionsThe anti-pyroptosis effect of AS-IV via the TXNIP-NLRP3-GSDMD axis improves the renal function and podocyte damage of db/db mice and delays the onset of DKD, according to in vivo and in vitro experimental data.

Background: Patients with type 2 diabetes mellitus (T2DM) pass through pre-diabetic stages and during diagnosis, half of them may have different complications including diabetic nephropathy. Hypertension has adverse impacts on diabetic nephropathy. We aimed to evaluate the risk factors for diabetic nephropathy among newly detected T2DM patients with normal blood pressure.
 Methods: A case-control study was done at a tertiary care hospital in Dhaka, Bangladesh from January 2018 to June 2019. Newly detected (<3 months), adult ( ³18 years), T2DM patients, who underwent tests for urine albumin-to-creatinine ratio (UACR), at least twice, 6-weeks apart, were included in this study. Patients with hypertension, known kidney disease, features of glomerulonephritis, systemic diseases including systemic lupus erythematosus and vasculitis, history of recent fever and exercise, urinary tract infection and pregnancy were excluded. Patients with UACR ³30 mg/g in at least two samples were cases and those with UACR <30 mg/g were controls.
 Results: Among the total 135 patients, 27 were cases [moderately increased proteinuria (previously, microalbuminuria) (UACR 30–299 mg/g) = 25 and severely increased proteinuria (previously, overt proteinuria) (UACR ³300 mg/g) = 2] and 108 were controls. Multivariate logistic regression revealed that family history of diabetes (OR=2.13; 95% CI, 1.412 to 4.216, p=0.003) and diabetic nephropathy (OR=3.31; 95% CI, 2.145 to 5.249, p=0.001), smoking (OR=3.11; 95% CI, 2.234 to 4.123, p=0.003), dyslipidaemia (OR=2.14; 95% CI, 1.363 to 3.324, p=0.005) and diabetic retinopathy (OR=6.23; 95% CI, 4.197 to 9.464, p=0.004) were significant risk factors.
 Conclusion: We concluded that, family history of diabetes and diabetic nephropathy, smoking, dyslipidaemia and diabetic retinopathy appeared as significant risk factors for diabetic nephropathy among newly diagnosed T2DM patients with normal blood pressure.
 BIRDEM Med J 2022; 12(2): 152-157

Background and Aims Diabetes mellitus (DM) is the leading cause of chronic kidney disease; principally resulting from the increasing prevalence of type 2 DM (T2DM). Patients with T2DM pass through pre-diabetic stages and at the time of diagnosis, up to half of the T2DM patients may have different macro- and micro-vascular complications, including diabetic nephropathy. Hypertension has adverse impacts on diabetic nephropathy. This study was designed to evaluate the risk factors for diabetic nephropathy among newly detected T2DM patients with normal blood pressure. Method This case-control study was done at out-patient department of a referral hospital in Dhaka, Bangladesh from January 2018 to June 2019. Newly detected (<3 months), adult (≥18 years), T2DM patients of either sex, who underwent test for urine albumin-to-creatinine ratio (UACR), at least twice (6 weeks apart), within a 6-month period, were included in this study. Patients with hypertension (newly diagnosed or known cases/on antihypertensive medications), known kidney disease, features of glomerulonephritis, systemic diseases including systemic lupus erythematosus and vasculitis, history of recent fever and exercise, urinary tract infection and pregnancy were excluded. Patients with UACR ≥30 mg/g in at least two (of three, if done) samples were cases and those with UACR <30 mg/g were controls. Results Total patients were 135, including 27 cases [moderately increased proteinuria (previously, microalbuminuria) (UACR 30 – 299 mg/g) = 25 and severely increased proteinuria (previously, overt proteinuria) (UACR ≥300 mg/g) = 2] and 108 controls. Mean age was 42.9 years and there was female (64.8%) predominance. Thirteen percent patients were smokers, 12% had dyslipidaemia, 51% had family history of DM and 44.9% had family history of diabetic nephropathy. Besides diabetic nephropathy, other chronic complications of DM were diabetic retinopathy (6.7%), diabetic peripheral neuropathy (0.7%) and coronary artery disease (1.5%). Regarding risk factors for diabetic nephropathy, family history of DM [odds ratio (OR) = 2.31, 95% confidence interval (CI) = 0.923 – 5.415, p = 0.003) and diabetic nephropathy (OR = 3.28, 95% CI = 1.523 – 9.297, p = 0.001), smoking (OR = 3.08, 95% CI = 1.066 – 0.934, p = 0.003), dyslipidaemia (OR = 2.11, 95% CI = 0.241 – 3.464, p = 0.004) and coexisting diabetic retinopathy (OR = 6.51, 95% CI = 1.46 – 11.79, p = 0.003) were significant. On multivariate logistic regression, family history of DM (OR = 2.13, 95% CI = 1.412 – 4.216, p = 0.003) and diabetic nephropathy (OR = 3.31, 95% CI = 2.145 – 5.249, p = 0.001), smoking (OR = 3.11, 95% CI = 2.234 – 4.123, p = 0.003), dyslipidaemia (OR = 2.14, 95% CI = 1.363 – 3.324, p = 0.005) and diabetic retinopathy (OR = 6.23, 95% CI = 4.197 – 9.464, p = 0.004) were significant. Conclusion Family history of DM and diabetic nephropathy, smoking, dyslipidaemia and concomitant diabetic retinopathy were significant risk factors for diabetic nephropathy among newly diagnosed T2DM patients with normal blood pressure.

This study aimed to investigate the correlation between serum miR-154-5p and urinary albumin to creatinine ratio (UACR) in patients with type 2 diabetes mellitus (T2DM) and the association with biomarkers of inflammation and fibrosis in diabetic kidney disease (DKD). A total of 390 patients with T2DM were divided into three groups: normal albuminuria (UACR < 30 mg/g, n = 136, NA), microalbuminuria (UACR at 30-300 mg/g, n = 132, MA), and clinical albuminuria (UACR > 300 mg/g, n = 122, CA). Circulating miR-154-5p, inflammatory (C-reactive protein (CRP); erythrocyte sedimentation rate (ESR); and tumor necrosis factor-α (TNF-α) and fibrotic markers (vascular endothelial growth factor (VEGF); transforming growth factor-β1 (TGF-β1); and fibronectin (FN)), and other biochemical indicators were assessed via real-time PCR, enzyme-linked immunosorbent assay, and chemiluminescence assay in patients with T2DM and 138 control subjects (NC). UACR, miR-154-5p, glycated hemoglobin (HbA1c), serum creatinine (sCr), blood urea nitrogen (BUN), ESR, CRP, VEGF, TNF-α, TGF-β1, and FN were significantly higher and the estimated glomerular filtration rate (eGFR) was significantly lower in NA, MA, and CA groups than in NC subjects (P < 0.05). Elevated levels of UACR and miR-154-5p were directly correlated with HbA1c, sCr, BUN, ESR, CRP, VEGF, TNF-α, TGF-β1, and FN and negatively correlated with eGFR (P < 0.05). miR-154-5p, HbA1c, sCr, BUN, eGFR, ESR, CRP, VEGF, TNF-α, TGF-β1, and FN were important factors affecting UACR. These findings indicated that elevated serum miR-154-5p is significantly correlated with high UACR in patients with T2DM and may offer a novel reference for the early diagnosis of DKD.

The aim is to investigate the correlations of serum retinol-binding protein (RBP) and stromal cell-derived factor-1 (SDF-1) with renal function in patients with diabetic kidney disease (DKD). A total of 438 patients with type 2 diabetes mellitus (T2DM) treated from October 2017 to October 2020 were enrolled in this prospective study and divided into simple T2DM and DKD groups. According to urinary albumin-to-creatinine ratio (UACR), DKD patients were divided into moderate, severe, and nephrotic groups. They were assigned to one of the following categories of estimated glomerular filtration rate (eGFR): G1, G2, G3a, G3b, G4, and G5 stages. The correlations of RBP and SDF-1 with renal function were analyzed. The DKD group had a longer T2DM course and higher RBP, uric acid (UA), blood urea nitrogen (BUN), β2-microglobulin (β2-MG), serum creatinine (Scr) levels and UACR, and lower SDF-1 level and eGFR than those of simple T2DM group (p < 0.05). The areas under the receiver operating characteristic curves of RBP and SDF-1 for identifying DKD were 0.903 and 0.868, and the optimal cutoff values were 70.71 mg/L and 5.69 ng/mL, respectively. With increasing urinary albumin and clinical stage, RBP, UA, BUN, β2-MG and Scr levels and UACR significantly rose, while SDF-1 level and eGFR declined (p < 0.05). In patients with DKD, RBP was correlated positively with UACR, UA, BUN, β2-MG, and Scr (r = 0.764/0.787/0.693/0.577/0.801, p < 0.0001), and negatively with eGFR (r = -0.782, p < 0.0001). SDF-1 was correlated negatively with UACR, UA, BUN, β2-MG and Scr (r = -0.744/-0.794/-0.666/-0.605/-0.820, p < 0.0001), and positively with eGFR (r = 0.767, p < 0.0001). The multiple linear regression equation was RBP = 29.852 + 0.007 x UACR + 0.101 x UA + 0.497 x BUN + 0.034 x Scr-0.083 x eGFR (p < 0.001). RBP and SDF-1 can identify DKD in patients with T2DM, and the degree of renal function damage is correlated positively with RBP and negatively with SDF-1. Elevated levels of UA, BUN, Scr and UACR as well as reduced eGFR are risk factors for evaluating RBP.