Diabetic Microangiopathy Research Articles

Association between Circulating Levels of 25-Hydroxyvitamin D3 and Matrix Metalloproteinase-10 (MMP-10) in Patients with Type 2 Diabetes.

Background: Matrix metalloproteinase-10 (MMP-10) levels increase progressively starting from early diabetic kidney disease (DKD) stages. Vitamin D3 (vitD3) deficit is associated with a higher risk of diabetic microangiopathy. Reduced MMP-10 expression has been observed after exposure to vitD3. Aim: to assess how vitD3 status is related to MMP-10 levels in patients with Type 2 diabetes (T2D). Methods: 256 patients with T2D were included in this cross-sectional study. Demographic, clinical and serum MMP-10 and 25-hydroxyvitamin D3 (25(OH)D3) levels were collected from each patient. The association between MMP-10 and (25(OH)D3) levels was assessed using a correlation analysis and fitting a multivariate linear regression model. Results: Serum MMP-10 levels were inversely correlated with circulating 25(OH)D3 (rho = −0.25; p < 0.001). In the subgroup analysis this correlation was significant in patients with DKD (rho = −0.28; p = 0.001) and in subjects with vitD3 deficit (rho = −0.24; p = 0.005). In the regression model adjusted for kidney function, body adiposity, smoking and vitD supplementation MMP-10 levels were 68.7 pg/mL lower in patients with 25(OH)D3 > 20 ng/mL, with respect to ≤20 ng/mL (p = 0.006). Conclusions: vitD3 repletion status is an independent predictor of MMP-10 levels in T2D patients. Perhaps, high 25(OH)D3 values should be targeted in these patients in order to prevent vascular complications.

DRDB: A Machine Learning Platform to Predict Chemical-Protein Interactions towards Diabetic Retinopathy.

Diabetic retinopathy (DR), a diabetic microangiopathy caused by diabetes, affects approximately 93 million people, worldwide. However, the drugs used to treat DR have limited efficacy and the variety of side effects. This is possibly because the complicated pathogenesis of DR is associated with multiple proteins. In this work, we attempted to identify potential drugs against DR-associated proteins and predict potential targets for drugs using in silico prediction of chemical-protein interactions (CPI) based on multitarget quantitative structure-activity relationship (mt-QSAR) method. Therefore, we developed 128 binary classifiers to predict the CPI for 15 DR targets using random forest (RF), k-nearest neighbours (KNN), support vector machine (SVM), and neural network (NN) algorithms with MACCS, extended connectivity fingerprints (ECFP6) fingerprints, and protein descriptors. In order to facilitate discovery of the novel drugs and target identification using the 128 binary classifiers, a free web server (DRDB) was developed. Compound Danshen Dripping Pills (CDDP), composed of Salvia miltiorrhiza, Panax notoginseng, and borneol, is commonly used in the treatment of cardiovascular diseases. To explore the applicability of DRDB, the potential CPIs of CDDP in treatment of DR were investigated based on DRDB. In vitro experimental validation demonstrated that cryptotanshinone and protocatechuic acid, two key components of CDDP, are capable of targeting ICAM-1 which is one of the key target of DR. We hope that this work can facilitate development of more effective clinical strategies for the treatment of DR.

Signaling Pathways Related to Oxidative Stress in Diabetic Cardiomyopathy.

Diabetes is a chronic metabolic disease that is increasing in prevalence and causes many complications. Diabetic cardiomyopathy (DCM) is a complication of diabetes that is associated with high mortality, but it is not well defined. Nevertheless, it is generally accepted that DCM refers to a clinical disease that occurs in patients with diabetes and involves ventricular dysfunction, in the absence of other cardiovascular diseases, such as coronary atherosclerotic heart disease, hypertension, or valvular heart disease. However, it is currently uncertain whether the pathogenesis of DCM is directly attributable to metabolic dysfunction or secondary to diabetic microangiopathy. Oxidative stress (OS) is considered to be a key component of its pathogenesis. The production of reactive oxygen species (ROS) in cardiomyocytes is a vicious circle, resulting in further production of ROS, mitochondrial DNA damage, lipid peroxidation, and the post-translational modification of proteins, as well as inflammation, cardiac hypertrophy and fibrosis, ultimately leading to cell death and cardiac dysfunction. ROS have been shown to affect various signaling pathways involved in the development of DCM. For instance, OS causes metabolic disorders by affecting the regulation of PPARα, AMPK/mTOR, and SIRT3/FOXO3a. Furthermore, OS participates in inflammation mediated by the NF-κB pathway, NLRP3 inflammasome, and the TLR4 pathway. OS also promotes TGF-β-, Rho-ROCK-, and Notch-mediated cardiac remodeling, and is involved in the regulation of calcium homeostasis, which impairs ATP production and causes ROS overproduction. In this review, we summarize the signaling pathways that link OS to DCM, with the intention of identifying appropriate targets and new antioxidant therapies for DCM.

Correlation of serum D-dimer levels with stages of diabetic retinopathy and gycosylated hemoglobin levels

Diabetic retinopathy is a disorder where the perfusion of the retina may be affected owing to the microvascular changes taking place in the retinal blood vessels due to hypergycemia. D-dimer level has been used as a biomarker of hypercoagulability and fibrinolytic activity since it is a product of fibrin degradation. D-dimer level (as a marker of coagulation cascade/fibrinolysis activation) assessment in type 1 and type 2 diabetics and its correlation with diabetic retinopathy stages have shown positive correlation. The findings in present study endorse the view that Elevated D dimer levels in severe forms of diabetic retinopathy. 26.9% patients with D dimer values more than 1000 have had severe NPDR to proliferative diabetic retinopathy, while 80% patients with normal values have shown to have no diabetic changes in fundus. Poor glycemic control (&amp;#62;6.5%) becomes a strong predictor for occurrence of severe form of diabetic retinopathy. 92.3% patients have shown the same. Hence, this study showed a positive correlation between increased blood sugar levels, elevated glyco Hb levels and severity of diabetic retinopathy. Hence, prompt understanding and treatment of hyperglycemia may prevent complications of diabetic retinopathy. The levels of Plasma D dimer is higher in type 2 diabetes mellitus complicated with microangiopathy. D-dimer is an important marker for thrombus formation hence may play an important role in the pathogenesis of diabetic microangiopathy. Prophlylactic anticoagulant therapy and strict diabetic control can help in halting the progression of diabetic retinopathy.

Spirometry and Diffusion Studies in Patients with Type-2 Diabetes Mellitus and Their Association with Microvascular Complications

Diabetes is a systemic disease with-well known complications involving eyes, kidneys and nerves. The presence of an extensive pulmonary microvascular circulation and abundant connective tissue raises the possibility that lung may also be a target organ in diabetes. A total of 45 subjects were included in the study. All patients were evaluated for diabetic microangiopathies: nephropathy (by 24-hour protein excretion), retinopathy (by direct ophthalmoscopy) and neuropathy (by clinical examination). The patients were divided into following three groups: Group A: patients with type-2 diabetes mellitus (DM) with evidence of microangiopathy (n=15); Group B: patients with type-2 DM without any evidence of microangiopathy (n=15); Group C: non-diabetic subjects (n=15) as controls. Glycosylated haemoglobin (HbA1C) was measured as an indicator of glycemic control. Spirometry and single-breath diffusion capacity for carbon-monoxide (DLCO) were performed on all patients using Elite Series Body Plethysmograph machine. A significant reduction of diffusion capacity corrected for alveolar volume (%DL/VA) was observed in group A (p < 0.001), as compared to the other groups. There were no differences among the three groups for other pulmonary functions. There was a significant correlation between DL/VA percent predicted and albuminuria (r= -0.975, p < 0.001), and DL/VA percent predicted and the retinopathy (r = -0.550, p < 0.05). This study shows a mild reduction in diffusing capacity in patients with type-2 DM with microangiopathy.

Down-regulation of LCN2 attenuates retinal vascular dysfunction and caspase-1-mediated pyroptosis in diabetes mellitus.

BackgroundDiabetic retinopathy (DR) is a diabetic microangiopathy with increasing incidence, which seriously threatens the quality of life of patients. This study investigated the molecular regulation mechanism of lipocalin-2 (LCN2) in DR by targeting the function of human retinal vascular endothelial cells (HRVECs).MethodsThe expression of LCN2 in the retinal tissue of diabetic and high glucose (HG)-induced HRVECs was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis and western blotting assay. After intravitreal injection of adeno-associated virus (AAV)-NC or AAV-sh-LCN2, in vivo experiments, hematoxylin and eosin (H&E) staining, and retinal trypsin digestion experiments were performed to analyze the effect of LCN2 silencing on DR retinal tissue. Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) staining was used to evaluate apoptosis and immunohistochemical (IHC) staining was performed to detect the expressions of caspase-1. Western blot was used to detect the expressions of pyroptosis-associated proteins. After transfection of sh-NC and sh-LCN2, the function of HRVECs cells induced by HG was evaluated by wound healing assay, Transwell assay, and tube formation assay.ResultsThe expression of LCN2 was significantly up-regulated in diabetic retinal tissue and HG-induced HRVECs. In vivo experiments showed that LCN2 silencing can significantly reduce diabetic retinal injury. Cell function experiments also revealed that LCN2 silencing inhibited cell migration, invasion, and angiogenesis. Flow cytometry and immunofluorescence staining showed that downregulation of LCN2 could inhibit caspase-1 mediated pyroptosis in HG-induced HRVECs.ConclusionsDown-regulation of LCN2 can significantly inhibit cell migration, invasion, and angiopoiesis, and pyroptosis regulated by caspase-1, thus attenuating the progression of DR.

FC 126: Association Between Circulating Levels of 25-Hydroxyvitamin D and Metalloproteinase-10 (MMP 10) in Patients With Type 2 Diabetes Mellitus. Cross-Sectional Study

Abstract BACKGROUND AND AIMS Matrix metalloproteinases (MMP) play an important role in diabetic kidney disease (DKD) (1). MMP-10 levels have been shown to increase progressively starting from the early stages of DKD (2). Vitamin D (vitD) deficit is associated with a higher risk of microangiopathy in diabetes. In humans, vitD status correlates inversely with MMP-9 circulating levels and VitD supplementation is associated with a decrease in these levels (3). In vitro, studies demonstrated reduced expression of MMP-10 after an exposure to vitD (4). However, no studies have yet assessed how vitD status is related to MMP-10 circulating levels. We aimed (1) to evaluate the association between the serum levels of 25-hydroxyvitamin D [25(OH)D3] and MMP-10 in a cohort of type 2 diabetes (T2D) patients and (2) to study a potential effect of chronic kidney disease (CKD) stage or estimated glomerular filtration rate (eGFRcr) on this association. METHOD The cross-sectional association between MMP-10 and 25(OH)D3 levels (&amp;gt;30 ng/mL, normal; 20–30 ng/mL, insufficiency; &amp;lt;20 ng/mL, deficiency; and relevant cut-offs 45 and 15 ng/mL) was investigated, by multivariate linear regression analysis after MMP-10 logarithm transformation, in 256 T2D patients recruited (2009–2016) in the Clinica Universidad de Navarra (Spain). Study protocol was approved by the Ethical Committee (2021.183TFG). Covariates were smoking habit, body adiposity percentage estimated by CUN-BAE formula, CKD stage and vitD3 supplement use. Age and sex were already included in the CKD-EPI formula. Interactions and collinearities were also analysed. Statistical analysis was performed using STATA software and two-tailed P-values &amp;lt;0.05 were considered statistically significant. RESULTS Characteristics of patients are given in Table 1. VitD deficit was significantly (P &amp;lt;0.001) more prevalent in patients with CKD (61%) compared with those without CKD (40%). MMP-10 showed a significant negative correlation with 25(OH)D3 (rho = –0.246; P &amp;lt;0.001), which remained significant including only CKD patients (rho = –0.283; P &amp;lt;0.001) and stronger above 45 ng/mL (rho = –0.7308, P &amp;lt;0.001) or moderate between 30 and 45 ng/mL (rho = –0.3290, P = 0.0045) (Fig. 1). In the fully adjusted model, MMP-10 was negatively associated (P = 0.019) with severe vitD deficiency [estimated coefficient –0.86 (–0.76 to –0.97)]. There was no interaction between covariates and 25(OH)D levels and collinearities were also discarded. CONCLUSION In T2D patients, circulating MMP-10 was inversely associated with serum 25(OH)D3 levels, especially in those patients with CKD and at the extremes of vitD level range. These results suggest that higher values of 25(OH)D3 should be targeted in these patients. The efficacy of vitD supplementation in reducing circulating MMP-10 levels should be further examined.

MO631: Diabetic Retinopathy and/or Diabetic Nephropathy Confers A Worse Renal Prognosis

Abstract BACKGROUND AND AIMS Around 50%–60% lesions of non-diabetic nephropathy (NDN) have been identified in patients with diabetes and renal biopsy. NDN patients have a better renal prognosis and survival. Diabetic retinopathy (DR) is a microvascular complication of diabetes frequently associated with diabetic nephropathy (DN). The objective of the work is to study the renal prognosis and survival of patients with DN with or without DR. METHOD Retrospective, observational, multicenter study of the pathological results of diabetic patients with renal biopsy was undertaken. A total of 18 Spanish centers have participated. RESULTS Cohort of 832 patients: 621 men (74.6%), median age 61.7 ± 12.8 years, median creatinine 2.8 ± 2.2 mg/dL and proteinuria 2.7 (1.2–5.4) g/24 h. Of them, 26.6% (n = 221) presented DR, 18.8% (n = 156) peripheral vasculopathy and 17.7% (n = 147) ischemic heart disease. On biopsy, 39.5% (n = 329) of patients had DN, 413 (49.6%) NDN and 90 (10.8%) mixed forms. We evidenced more DR in the DN group compared with the other two groups (44.1% versus 11.1% in NDN and 33.3% in mixed forms). Regarding renal prognosis, 58.9% (n = 109) of patients with DR and DN and mixed forms required RRT versus 40.7% (n = 88) of patients without DR. A total of 39.1% (n = 18) of patients with DR and NDN required renal replacement therapy (RRT) versus 23.9% (n = 79) of patients without DR. Regarding mortality, 28.6% (n = 50) of patients with DR and DN or mixed forms died during the follow-up versus 22.7% (n = 49) of patients without DR. A total of 15.2% (n = 7) of patients with DR and NDN died and 15.4% (n = 51) of patients without DR. In survival analysis (Kaplan—Meier curves), patients with DR had worse renal prognosis (P &amp;lt; 0.001), patients with DN (P &amp;lt; 0.001) and patients with DR and/or DN and mixed forms (P = 0.02). Cox regression analysis identified as independent risk factors of RRT: age (OR 1.014;1.025–1.004, P = 0.006), creatinine (OR 1.262; 1.217–1.309; P &amp;lt; 0.001), proteinuria (OR 1.057; 1.03–1.085; P &amp;lt; 0.001), DR (OR 1.359; 1.007–1.833; P = 0.045), DR and/or DN (OR 1.59; 2.42–1.04; P = 0.033) and DN (OR1.489; 1.05–2.11; P = 0.026). CONCLUSION The presence of DR and/or DN confers a worse renal prognosis. The diagnosis of diabetic microangiopathy in terms of retinopathy and/or diabetic nephropathy is crucial since it confers a worse renal prognosis indicating patients at risk of progression to end-stage renal disease.

The Role of PKM2 in Diabetic Microangiopathy.

Diabetic microangiopathy is among the most common complications affecting patients with diabetes, and includes both diabetic retinopathy (DR) and diabetic nephropathy (DKD). Diabetic microangiopathy remains a persistent threat to the health and quality of life of affected patients. Mechanistically, the severity of DR and DKD is tied to mitochondrial and glucose metabolism abnormalities, with the activation of the glycolytic enzyme pyruvate kinase M2 (PKM2) contributing to mitochondrial and glomerular dysfunction, abnormal renal hemodynamics, and retinopathy. PKM2 can activate inflammatory bodies in macrophages to promote the release of inflammatory mediators, and serves as a key regulator of inflammatory factors, chemokines and adhesion molecules. As such, there is sufficient evidence that PKM2 can be used as a biomarker for the diagnosis of diabetes and diabetic microangiopathy. Here, we survey the mechanisms whereby PKM2 contributes to diabetes-related microvascular diseases, associated regulatory roles, post-translational modifications, and the potential utility of PKM2 as a therapeutic target. Through this literature review, we have determined that PKM2 offers promise as both a diagnostic marker and therapeutic target with direct relevance to research pertaining to diabetic microangiopathy.

Protective Effect and Possible Mechanisms of Artemisinin and Its Derivatives for Diabetic Nephropathy: A Systematic Review and Meta-Analysis in Animal Models.

Background Artemisinin and its derivatives have potential antidiabetic effects. There is no evaluation of reported studies in the literature on the treatment of diabetic nephropathy (DN), one of the commonest diabetic microangiopathies, with artemisinins. Here, we aimed to evaluate preclinical evidence for the efficacy and possible mechanisms of artemisinins in reducing diabetic renal injury. Methods We conducted an electronic literature search in fourteen databases from their inception to November 2021. All animal studies assessing the efficacy and safety of artemisinins in DN were included, regardless of publication or language. Overall, 178 articles were screened according to predefined inclusion and exclusion criteria. Finally, 18 eligible articles were included in this systematic review. The SYstematic Review Center for Laboratory animal Experimentation (SYRCLE) risk-of-bias tool was used to assess the risk of bias in the included studies. The primary outcomes were kidney function, proteinuria, and renal pathology. Secondary endpoints included changes in fasting plasma glucose (FPG) levels, body weight, and relevant mechanisms. Results Of the 18 included articles involving 418 animal models of DN, 1, 2, 6, and 9 used dihydroartemisinin, artemether, artesunate, and artemisinin, respectively. Overall, artemisinins reduced indicators of renal function, including blood urea nitrogen (P < 0.00001), serum creatinine (P < 0.00001), and kidney index (P = 0.0001) compared with control group treatment. Measurements of proteinuria (P < 0.00001), microalbuminuria (P < 0.05), and protein excretion (P = 0.0002) suggested that treatment with artemisinins reduced protein loss in animals with DN. Artemisinins may lower blood glucose levels (P = 0.01), but there is a risk of weight gain (P < 0.00001). Possible mechanisms of action of artemisinins include delaying renal fibrosis, reducing oxidative stress, and exerting antiapoptotic and anti-inflammatory effects. Conclusion Available evidence suggests that artemisinins may be protective against renal injury secondary to diabetes in preclinical studies; however, high-quality and long-term trials are needed to reliably determine the balance of benefits and harms.

Diabetes is an Independent Growth Factor of Ischemic Stroke During Reperfusion Phase Leading to Poor Clinical Outcome

ObjectivesDespite the success of recanalization by bridging therapy, about half of treated stroke patients remain disabled. While numerous reports propose clinical predictors of stroke clinical outcome in this context, we originally aimed to study pre-therapeutic factors influencing infarct growth (IG) and poor clinical outcome in strokes due to large vessel occlusion (LVO) successfully recanalized. Materials and methodsWe enrolled 87 consecutive successfully recanalized patients (mTICI: 2b/2c/3) by mechanical thrombectomy (±rt-PA) after stroke due to middle cerebral artery (M1) occlusion within 6 h according to AHA guidelines. IG was defined by subtracting the initial DWI volume to the final 24 h-TDM volume. Statistical associations between poor clinical outcome (mRS≥2), IG and pertinent clinico-radiological variables, were measured using logistic and linear regression models. ResultsAmong 87 enrolled patients (Age(y): 68.4 ± 17.5; NIHSS: 16.0 ± 5.4), 42/87 (48,28%) patients had a mRS ≥ 2 at 3 months. Diabetic history (OR: 3.70 CI95%[1.03;14.29] and initial NIHSS (/1 point: OR: 1.16 CI95%[1.05;1.27]) were independently associated with poor outcome. IG was significantly higher in stroke patients with poor outcome (+7.57 ± 4.52 vs −7.81 ± 1.67; p = 0.0024). Initial volumes were not significantly different (mRS≥2: 16.18 ± 2.67; mRS[0–1]: 14.70 ± 2.30; p = 0.6771). Explanatory variables of IG in linear regression were diabetic history (β: 21.26 CI95%[5.43; 37.09]) and NIHSS (β: 0.83 CI95%[0.02; 1.64]). IG was higher in diabetic stroke patients (23.54 ± 1.43 vs −6.20 ± 9.36; p = 0.0061). ConclusionsWe conclude that diabetes leads to continued IG after complete recanalization, conditioning clinical outcome in LVO strokes successfully recanalized by bridging therapy. We suggest that poor tissular reperfusion by diabetic microangiopathy could explain this result.

RNA-binding proteins in diabetic microangiopathy.

BackgroundAs the most common complication of diabetes, the diabetic microangiopathy characterizes diabetic retinopathy (DR) and nephropathy (DN). Diabetic microangiopathy has always been a serious clinical problem. A wide variety of nucleic acid interacting factors called the RNA binding proteins (RBPS) take part in several crucial cellular processes.MethodsOver the past decade, studies have shown that RBPs have crucial part in both malignant tumors and diabetes, especially in diabetic microangiopathy. This review examined the research history of RBPS in DR and DN.ResultsWe reviewed the literature and found that RBPS is potentially useful as therapeutic targets, diagnostic markers, or predict disease progression.ConclusionHuR acts as a vital therapeutic targeting protein in diabetic microangiopathy. IGF2BP2, P311, TTP, YBX1, and MBNL1 have a potential role in the treatment of DN.

Monocyte to high-density lipoprotein cholesterol ratio and serum uric acid in Chinese adults: a cross-sectional study

BackgroundPrevious studies have shown that the monocyte to high-density lipoprotein cholesterol (HDL-C) ratio (MHR) is a predictor of various diseases such as coronary heart disease, diabetic microangiopathy, and metabolic syndrome. However, there are few scientific reports on the correlation between MHR and serum uric acid. The objective of this report is to explore the relationship between MHR and serum uric acid in Chinese adults.MethodsThis cross-sectional study included 646 participants from southwest China who underwent a health examination at the Health Management Center of Deyang People’s Hospital. The examination included blood pressure readings, routine blood tests (lipid, fasting glucose, serum transaminase, and serum uric acid levels), and various standardized questionnaires. We employed a generalized additive model and smoothed curve fitting to explore the relationship between MHR and serum uric acid levels. We then performed subgroup analyses to investigate the robustness of this relationship.ResultsAfter adjusting for confounders (age, sex, body mass index, systolic blood pressure, diastolic blood pressure, aspartate transaminase, alanine aminotransferase, fasting glucose, total cholesterol, low-density lipoprotein, smoking, drinking, and exercise status), MHR was found to be positively correlated with serum uric acid levels (P < 0.001). The smoothing curve showed an approximately linear correlation between MHR and serum uric acid levels, and the linear correlation coefficient was 146.74 (95% CI 96.16–197.33, P < 0.0001). The subgroup analyses showed that the effect of MHR on serum uric acid levels was smaller in occasional smokers and smokers than in nonsmokers (P = 0.0194).ConclusionMHR was significantly and positively correlated with serum uric acid levels. Additionally, the effect of MHR on serum uric acid levels was lower in the individuals who smoked more.

Clinical significance and influencing factors of carotid pulse wave velocity in patients with diabetic microangiopathy.

To evaluate the utility of carotid ultrafast pulse wave velocity (PWV) and explore its influencing factors in patients with type 2 diabetes mellitus (T2DM) microangiopathy. Seventy-seven patients with T2DM were divided into two groups according to the absence (Group A, n=45) or presence (Group B, n=32) of microangiopathy. The control group comprised 1544 healthy volunteers. Two-dimensional ultrasonography was used to measure intima-media thickness (IMT) of the carotid arteries, and ultrafast ultrasound imaging was used to measure PWV of the carotid arteries at the beginning of systole (PWV-BS) and the end of systole (PWV-ES). The IMT, PWV-BS, and PWV-ES were higher in the T2DM group than in the control group, and the values in T2DM Group B were higher than those in Group A. IMT was positively correlated with PWV-BS and PWV-ES. Age and uric acid were influencing factors of PWV-ES, while age, uric acid, body mass index, glycated hemoglobin, and urine albumin/creatinine ratio were influencing factors of PWV-BS. PWV-ES was a more sensitive predictor than PWV-BS, and a PWV-ES critical value predicted carotid elasticity in patients with T2DM microangiopathy. Ultrafast PWV can reflect early atherosclerosis and provide a noninvasive assessment of microangiopathy in patients with T2DM.

Correlation of Albuminuria and Diabetic Retinopathy in Type-II Diabetes Mellitus Patients

Background: Diabetic ocular disease is a leading cause of blindness today. The most common microvascular complications of diabetes are diabetic retinopathy and diabetic nephropathy. Multiple risk factors like the duration of the disease, age of the patient, high blood pressure, pregnancy, blood glucose control, and nephropathy have been studied to be associated with the development and progression of diabetic microangiopathy. However, the association of albuminuria has still not been studied in detail, especially in type-II diabetes mellitus.Aim: The primary objective of our study is to quantify the relationship between diabetic retinopathy and urine albumin excretion and to correlate the urinary albumin excretion (normoalbuminuria, microalbuminuria, macroalbuminuria) with the severity and grade (mild, moderate, severe non-proliferative diabetic retinopathy [NPDR] or proliferative diabetic retinopathy [PDR]) of diabetic retinopathy.Methods: In this cross-sectional study, 250 patients with type-II diabetes above 40 years of age attending the ophthalmic outpatient department (OPD), Kalinga Institute of Medical Sciences (KIMS), Bhubaneswar in India between September 2019 and September 2021 were subjected to a detailed evaluation of history and a thorough ocular examination. Besides, a blood sugar estimation and urine albumin levels were documented. The grade of diabetic retinopathy was correlated with albumin levels.Results: The duration of diagnosis of diabetes ranged from 1-25 years. The association between the grade of diabetic retinopathy and the duration since diagnosis was significant. Sixty-nine percent of the cases were hypertensives, and 66.7% of hypertensives had diabetic retinopathy. In patients without retinopathy, 83.03% had normoalbuminuria levels, and 16.96% had microalbuminuria. In the mild NPDR group, 37.94% of cases had normoalbuminuria, and 62.06% had microalbuminuria. In the moderate NPDR group, 11.1% of cases had normoalbuminuria, and 88.8% had microalbuminuria. In the severe NPDR group, 57.14% of cases had microalbuminuria, while 42.86% had macroalbuminuria. In the very severe NPDR group, 42.86% of cases had microalbuminuria, and 57.14% had macroalbuminuria. In the PDR group, only 6.6% of cases had microalbuminuria, and the rest, 93.3%, had macroalbuminuria.Conclusion: This study concluded that there is a definite association between albuminuria and severe diabetic retinopathy in type-II diabetes. Microalbuminuria was a finding associated with all grades of retinopathy with skewing towards the lower grades of diabetic retinopathy; a proportion of diabetics without retinopathy also had microalbuminuria, while macroalbuminuria was associated only with those patients who had either severe NPDR, very severe NPDR, or PDR.

A Study of the Protective Effect of Bushen Huoxue Prescription on Cerebral Microvascular Endothelia Based on Proteomics and Bioinformatics.

Diabetic cognitive dysfunction is a serious complication of type 2 diabetes mellitus (T2DM), which can cause neurological and microvascular damage in the brain. At present, there is no effective treatment for this complication. Bushen Huoxue prescription (BSHX) is a newly formulated compound Chinese medicine containing 7 components. Previous research indicated that BSHX was neuroprotective against advanced glycosylation end product (AGE)-induced PC12 cell insult; however, the effect of BSHX on AGE-induced cerebral microvascular endothelia injury has not been studied. In the current research, we investigated the protective effects of BSHX on AGE-induced injury in bEnd.3 cells. Our findings revealed that BSHX could effectively protect bEnd.3 cells from apoptosis. Moreover, we analyzed the network regulation effect of BSHX on AGE-induced bEnd.3 cells injury at the proteomic level. The LC-MS/MS-based shotgun proteomics analysis showed BSHX negatively regulated multiple AGE-elicited proteins. Bioinformatics analysis revealed these differential proteins were involved in multiple processes, such as Foxo signaling pathway. Further molecular biology analysis confirmed that BSHX could downregulate the expression of FoxO1/3 protein and inhibit its nuclear transfer and inhibit the expression of downstream apoptotic protein Bim and the activation of caspase, so as to play a protective role in AGE-induced bEnd.3 injury. Taken together, these findings demonstrated the role of BSHX in the management of diabetic cerebral microangiopathy and provide some insights into the proteomics-guided pharmacological mechanism study of traditional Chinese Medicine.

DR1 Activation Inhibits the Proliferation of Vascular Smooth Muscle Cells through Increasing Endogenous H2S in Diabetes.

Tissue ischemia and hypoxia caused by the abnormal proliferation of smooth muscle cells (SMCs) in the diabetic state is an important pathological basis for diabetic microangiopathy. Studies in recent years have shown that the chronic complications of diabetes are related to the decrease of endogenous hydrogen sulfide (H2S) in diabetic patients, and it has been proven that H2S can inhibit the proliferation of vascular SMCs (VSMCs). Our study showed that the endogenous H2S content and the expression of cystathionine gamma-lyase (CSE), which is the key enzyme of H2S production, were decreased in arterial SMCs of diabetic mice. The expression of PCNA and Cyclin D1 was increased, and the expression of p21 was decreased in the diabetic state. After administration of dopamine 1-like receptors (DR1) agonist SKF38393 and exogenous H2S donor NaHS, the expression of CSE was increased and the change in proliferation-related proteins caused by diabetes was reversed. It was further verified by cell experiments that SKF38393 activated calmodulin (CaM) by increasing the intracellular calcium ([Ca2+]i) concentration, which activated the CSE/H2S pathway, enhancing the H2S content in vivo. We also found that SKF38393 and NaHS inhibited insulin-like growth factor-1 (IGF-1)/IGF-1R and heparin-binding EGF-like growth factor (HB-EGF)/EGFR, as well as their downstream PI3K/Akt, JAK2/STAT3 and ERK1/2 pathways. Taken together, our results suggest that DR1 activation up-regulates the CSE/H2S system by increasing Ca2+-CaM binding, which inhibits the IGF-1/IGF-1R and HB-EGF/EGFR pathways, thereby decreasing their downstream PI3K/Akt, JAK2/STAT3 and ERK1/2 pathways to achieve the effect of inhibiting HG-induced VSMCs proliferation.

Carvedilol activates nuclear factor E2-related factor 2/ antioxidant response element pathway to inhibit oxidative stress and apoptosis of retinal pigment epithelial cells induced by high glucose

ABSTRACT Diabetic retinopathy (DR) is the most prominent manifestation of diabetic microangiopathy and is a serious complication of diabetes. Despite extensive researches focusing on DR, treatment options for DR are still limited. Carvedilol (CAR) has vasodilatory, antioxidant stress and anti-inflammatory effects and poses a vital role in addressing the issue of diabetic complications. This paper attempts to explore this property of CAR and investigate into its effects on DR. First, ARPE-19 cells were treated with different concentrations of CAR and cells were induced with 30 mM high glucose (HG) to establish a DR cell model. Cell viability was assayed by cell counting kit-8 (CCK-8) with or without HG induction. Cellular inflammation and oxidative stress were evaluated by enzyme-linked immunosorbent assay (ELISA) and corresponding kits. The measurement of apoptosis levels was conducted by Terminal dUTP nick-end labeling (TUNEL) and Western blotting. The protein levels related to Nrf2/ARE signaling pathway were assessed by Western blotting. Finally, cellular inflammation, oxidative stress and apoptosis in ARPE-19 cells pretreated with Nrf2 inhibitor ML385 were tested again by the same methods. Results showed that under HG induction, CAR effectively improved ARPE-19 cell viability, inhibited cellular inflammation, oxidative stress, and apoptosis. Moreover, CAR activated Nrf2/ARE signaling pathway, which further suppressed cellular inflammation, oxidative stress, and apoptosis. Overall, CAR inhibited HG-induced oxidative stress and apoptosis in retinal pigment epithelial cells by activating Nrf2/ARE pathway.

Age features of morphological changes of the hematotesticular barrier in experimental diabetes mellitus.

Background. Diabetes mellitus is an acute medical and social problem in all parts of the world due to the constant increase in the incidence of the disease, severe complications, disability and high mortality. Hyperglycemia leads to oxidative stress and increased processes of formation of active oxygen species, which in turn make a significant contribution to the development of male infertility. Objective. Therefore, the aim of our study was to establish morphological changes in the vessels of the hemomicrocirculatory flow and testicular sustentocytes of adult rats with experimental streptozotocin diabetes mellitus (SDM). Methods. The material for the study were the testicles of 20 sexually mature 6-month-old rats (weighing 150-180 g). SDM in animals of the experimental group was simulated by a single intraperitoneal injection of streptozotocin (dissolved in 0.1 M citrate buffer solution with a pH of 4.5) at a dose of 6 mg per 100 g of mass. An equivalent dose of 0.1 M citrate buffer was injected intraperitoneally to animals of the control group. Histological, electron microscopic, biochemical, morphometric and statistical research methods were used. Results. It was found that in the early stages of SDM (28th day) on the background of hyperglycemia in the hemomicrocirculatory flow of the testes there is a spasm of the vessels of the afferent link, which is confirmed by a decrease in the lumen of arterioles and an increase in their Wagenworth index. On the 70th day of SDM on the background of elevated levels of glucose and glycosylated hemoglobin in the links of the hemomicrocirculatory flow of the testis there are signs of diabetic microangiopathy, manifested by: hemorheological disorders in micro-hemo-vessels, (erythrocyte sludge, adhesion of erythrocytes and platelets, microclasmatosis), decrease in the capacity of arterioles and capillaries (increase in the Wagenworth index, respectively by 1.8 and 1.9 times), microclasmatosis, thickening and proliferation of the basement membrane of capillaries. Against the background of diabetic microangiopathy, there is a decrease in the number of sustentocytes by 0.01 mm2 of testicular parenchyma by 1.8 times, compared with control indicators, the area of their profile increases by 2.2 times (in all cases p&lt;0.05). The nucleolar areas probably do not change, which leads to an increase in the nuclear-cytoplasmic ratio by 2.9 times (p&lt;0.05). Such morphometric changes of sustentocytes are caused by development of vacuolar hydropic dystrophies in them, and an apoptosis that is confirmed by data of histo-ultrastructural studies. Changes in sustentocytes against the background of the development of diabetic microangiopathy lead to a violation of the hematotesticular barrier and to dystrophic changes in the spermatogenic epithelium of the testis. Conclusion. Thus, on the 70th day of SDM in the hemomicrocirculatory flow of the testis, the development of diabetic microangiopathy is observed, which leads to a violation of the hematotesticular barrier, and as a consequence, to a violation of spermatogenesis.

Necrobiosis lipoidica: a rare clinical and pathomorphological case

According to modern scientific researches, necrobiosis lipoidica (NL) is a disease characterized by focal disorganization and lipid collagen dystrophy. It is believed that the basis of skin changes in this dermatosis is diabetic microangiopathy that is accompanied by sclerosis and obliteration of blood vessels, which leads to necrobiosis with subsequent deposition of lipids in the dermis. This pathology is registered relatively rarely, in 1 % of patients with diabetes mellitus (DM) on average. The combination of NL with DM, according to the literature data, ranges from 25 to 70%; more often (in 40–60 % of cases) DM is preceded by NL, and in 10–25 % of cases they occur simultaneously. In addition, in 10–50 % of cases NL is diagnosed in people without concomitant diabetes. The variability of clinical, epidemiological features and the relatively low prevalence of this pathology is often the cause for misdiagnosis or late diagnosis. The described clinical case is typical in terms of the epidemiological data: sex, age, presence of DM. At the same time, it is rare in terms of the clinical picture: it is not classically diabetic by localization (symmetrical areas of the legs are typical), by appearance of necrobiosis areas — granulomatous type of necrobiosis in the form of granuloma annulare, by histological structure — area of chronic perivascular lymphoplasmocytic inflammation with the involvement of single giant cells, which required additional clinical and anamnestic data for an objective report of the pathologist. Biopsy in this case was used as a differential diagnosis between granuloma annulare and necrobiotic necrogranuloma. In addition, this method of diagnosis has played an additional therapeutic role. This case may have demonstrated the activation of the cellular and humoral immune response in the area of chronic inflammation in response to a mechanical damage and the resolution of inflammation with complete tissue repair.