Microvascular Complication Of Diabetes Mellitus Research Articles

Tocotrienol-rich fraction reduces retinal inflammation and angiogenesis in rats with streptozotocin-induced diabetes

BackgroundDiabetic retinopathy (DR) is the second commonest microvascular complication of diabetes mellitus. It is characterized by chronic inflammation and angiogenesis. Palm oil-derived tocotrienol-rich fraction (TRF), a substance with anti-inflammatory and anti-angiogenic properties, may provide protection against DR development. Therefore, in this study, we investigated the effect of TRF on retinal vascular and morphological changes in diabetic rats. The effects of TRF on the retinal expression of inflammatory and angiogenic markers were also studied in the streptozotocin (STZ)-induced diabetic rats.MethodsMale Sprague Dawley rats weighing 200–250 g were grouped into normal rats (N) and diabetic rats. Diabetes was induced by intraperitoneal injection of streptozotocin (55 mg/kg body weight) whereas N similarly received citrate buffer. STZ-injected rats with blood glucose of more than 20 mmol/L were considered diabetic and were divided into vehicle-treated (DV) and TRF-treated (DT) groups. N and DV received vehicle, whereas DT received TRF (100 mg/kg body weight) via oral gavage once daily for 12 weeks. Fundus images were captured at week 0 (baseline), week 6 and week 12 post-STZ induction to estimate vascular diameters. At the end of experimental period, rats were euthanized, and retinal tissues were collected for morphometric analysis and measurement of NFκB, phospho-NFκB (Ser536), HIF-1α using immunohistochemistry (IHC) and enzyme-linked immunosorbent assay (ELISA). Retinal inflammatory and angiogenic cytokines expression were measured by ELISA and real-time quantitative PCR.ResultsTRF preserved the retinal layer thickness (GCL, IPL, INL and OR; p < 0.05) and retinal venous diameter (p < 0.001). TRF also lowered the retinal NFκB activation (p < 0.05) as well as expressions of IL-1β, IL-6, TNF-α, IFN-γ, iNOS and MCP-1 (p < 0.05) compared to vehicle-treated diabetic rats. Moreover, TRF also reduced retinal expression of VEGF (p < 0.001), IGF-1 (p < 0.001) and HIF-1α (p < 0.05) compared to vehicle-treated rats with diabetes.ConclusionOral TRF provided protection against retinal inflammation and angiogenesis in rats with STZ-induced diabetes by suppressing the expression of the markers of retinal inflammation and angiogenesis.

1,8-Cineole ameliorates diabetic retinopathy by inhibiting retinal pigment epithelium ferroptosis via PPAR-γ/TXNIP pathways

1,8-Cineole, the main component of volatile oil in aromatic plants, has diverse pharmacological properties, including antioxidant, anti-inflammatory, and anti-cancer properties. Diabetic retinopathy (DR) is a common microvascular complication of diabetes mellitus (DM). Here, we investigated the protective effect of 1,8-cineole on DR and found that 1,8-cineole treatment could alter the expression of several genes in both high glucose (HG)-induced ARPE-19 cells and retinal tissues of DM mice, as well as inhibit ferroptosis. Subsequent investigations into the molecular mechanisms underlying this inhibition revealed that expression of thioredoxin-interacting protein (TXNIP) was significantly upregulated while that of peroxisome proliferator-activated receptor γ (PPAR-γ) was significantly downregulated in HG-induced ARPE-19 cells, and treatment with 1,8-cineole could effectively reverse these changes. Treatment with a PPAR-γ pharmacological agonist (rosiglitazone), alone or combined with 1,8-cineole, significantly inhibited the transcription of TXNIP and ferroptosis in HG-induced ARPE-19 cells. Conversely, pretreatment with GW9662, a PPAR-γ inhibitor, upregulated the transcription and expression of TXNIP in HG-induced ARPE-19 cells; 1,8-cineole failed to reverse this upregulated expression. To explore these relationships, we constructed a PPAR-γ adenovirus shRNA to elucidate the effect of 1,8-cineole on the negative regulation of TXNIP by PPAR-γ. Taken together, the present findings indicate that HG-induced ferroptosis in retinal tissue plays an essential role in the pathogenesis of DR, which can be ameliorated by 1,8-cineole.

EE350 Cost-Effectiveness of Mirogabalin in Treatment of Diabetic Peripheral Neuropathic Pain in Mainland China

Diabetic peripheral neuropathic pain (DPNP) is a chronic microvascular complication of diabetes mellitus associated with debilitating pain, high morbidity, and poor quality of life. The objective of this study was to compare the cost-effectiveness of mirogabalin, a potent and selective αlpha-2-delta ligand, with pregabalin in patients with DPNP from the healthcare system perspective in mainland China.

Metabolic Syndrome in Type 2 Diabetes Mellitus Patients: Prevalence, Risk Factors, and Associated Microvascular Complications.

Background The chronic macro and microvascular complications of diabetes mellitus pose serious health challenges. Metabolic syndrome (MetSy) is characterized by central obesity, glucose intolerance, hyperinsulinemia, low high-density lipoproteins (HDLs), high triglycerides (TGs), and hypertension. MetSy precedes or accompanies diabetes, and it has been linked to an increased risk of cardiovascular disease and premature death. This study aimed to estimate prevalence, identify risk factors, and evaluate associated microvascular complications among MetSy patients with type 2 diabetes mellitus (T2DM). Methodology Over the period of March 20, 2022, to March 31, 2023, a prospective cohort study was conducted at the Outdoor Clinic and Medicine Department of Sheikh Zayed Hospital, Rahim Yar Khan. Based on the International Diabetes Federation MetSy criteria, a total of 160 patients fulfilling the inclusion criteria were selected. A special proforma was used to obtain sociodemographic, clinical, and laboratory variables of MetSy in diabetic participants. Blood pressure and anthropometric measurements such as waist circumference (WC) and body mass index (BMI) were measured. Fasting venous blood was collected to analyze biochemical variables such as fasting blood sugar (FBS), TG, and high-density lipoprotein-cholesterol (HDL-C). The microvascular complications of T2DM were established using fundus ophthalmoscopy and neurological and kidney function assessments with the help oflaboratory tests. These variables were matched between MetSy and no MetSy groups along with the presence or absence of diabetes microvascular complications. This information was analyzed based on these assessments and patient interviews. Results Of the 160 T2DM patients, the mean age was 52 years with a predominance of females (51.8%) in the 50-59-year age group (56.8%). The average BMI for females was 29.38 ± 0.54 kg/m², and 32 (20%) had obesity. Females exhibited a large WC of 93.52 ± 1.58 cm, and 48 of 83 females had reported diabetes microvascular complications. A significant p-value was observed for hypertension, high TG, low HDL-C, large WC, obesity, BMI, age, and female gender on comparing diabetics with metabolic syndrome (MetSy+) and those without metabolic syndrome (MetSy-). The prevalence of microvascular complications in T2DM patients with MetSy+ was 52.5% compared with 47.5% in MetSy-. The prevalence of diabetic retinopathy was 24.9% (95% confidence interval (CI) = 20.3%-29.6%), nephropathy was 16.8% (95% CI = 12.8%-20.7%), and neuropathy was 10.8% (95% CI = 7.4%-13.3%). Conclusions The prevalence of MetSy observed among T2DM patients was 65%, with married obese females in the 50-59-year age group being more likely to be affected than males. Hypertension, poor glycemic control, high TG, low HDL-C, and greater anthropometric waist measurements and BMI were additional risk factors that tended to increase the MetSy burden in T2DM. Diabetic retinopathy, nephropathy, and neuropathy were the most prevalent microvascular complications of diabetes, and immediate attention is needed to stop their detrimental effects. Longer uncontrolled diabetes, increasing age, and hypertension were independent predictors of microvascular complications. To further reduce the risks of complications that threaten healthy aging and prognosis for these patients, MetSy screening, health education, and better diabetic management are crucial.

Coptisine mitigates diabetic nephropathy via repressing the NRLP3 inflammasome.

Diabetic nephropathy is a microvascular complication of diabetes mellitus, threatening the health of millions of people. Herein, we explored a blood glucose independent function of coptisine on diabetic nephropathy. A diabetic rat model was established by intraperitoneal administration of streptozotocin (65 mg/kg). Coptisine treatment (50 mg/kg/day) retarded body weight loss and reduced blood glucose. On the other hand, coptisine treatment also decreased kidney weight and the levels of urinary albumin, serum creatinine, and blood urea nitrogen, indicating an improvement of renal function. Treatment with coptisine also mitigated renal fibrosis, with alleviative collagen deposition. Likewise, in vitro study showed that coptisine treatment decreased apoptosis and fibrosis markers in HK-2 cells treated with high glucose. Furthermore, after coptisine treatment, the activation of NOD-like receptor pyrin domain containing protein 3 (NRLP3) inflammasome was repressed, with decreased levels of NLRP3, cleaved caspase-1, interleukin (IL)-1β, and IL-18, indicating that the repression of NRLP3 inflammasome contributed to the effect of coptisine on diabetic nephropathy. In conclusion, this study revealed that coptisine mitigates diabetic nephropathy via repressing the NRLP3 inflammasome. It is indicated that coptisine may have the potential to be used in the diabetic nephropathy treatment.

Withametelin, a steroidal lactone, isolated from datura innoxa attenuates STZ-induced diabetic neuropathic pain in rats through inhibition of NF-kB/MAPK signaling.

Diabetic neuropathic pain is one of the microvascular complications of diabetes mellitus characterized by symmetrical pain and sensory abnormalities. A steroidal lactone isolated from the datura innoxa plant, withametelin (WMT), exhibited significant neuroprotective, anti-inflammatory, antioxidant, and anticancer properties. The current study aimed to investigate anti-neuropathic pain activity and the molecular mechanism of WMT against streptozotocin (STZ)-induced diabetic neuropathy. Rats were given a single injection of STZ (60mg/kg, intraperitoneally (i.p.)) for induction of diabetes on the first day of the study. After the onset of diabetic neuropathy, pregabalin (10mg/kg, i.p.) and WMT (0.1 and 1mg/kg, i.p.) treatments were started from day 14 up to day 42. It was found that STZ-induced neuropathic pain behaviors were markedly reduced by WMT. It inhibited the STZ-associated histopathological changes and genotoxicity in the sciatic nerve and spinal cord. Additionally, Fourier transforms infrared (FTIR) spectroscopy results revealed that STZ-induced alterations in the biochemical components of the sciatic nerve's myelin sheath were inhibited by WMT. In the spinal cord, it markedly reduced the immunoreactivity of mitogen-activated protein kinases (MAPKs) signaling components such as p38-MAPK, c-Jun N-terminal kinase (JNK), extracellular-signal-regulated-kinase (ERK), and activator-protein 1 (AP-1). It also reduced the expression levels of nuclear factor-kappa-B (NF-κB), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS). The production of inflammatory cytokines was considerably reduced by WMT. This study provides convincing evidence that WMT treatment attenuated STZ-induced diabetic neuropathic pain by inhibition of MAPK/NF-κB signaling.

ADAM metallopeptidase domain 10 knockdown enables podocytes to resist high glucose stimulation by inhibiting pyroptosis via MAPK pathway.

Diabetic nephropathy (DN) is a common severe microvascular complication of diabetes mellitus, and podocyte damage occurs in the early stages of DN. The urine of patients with various types of glomerular disease presents increased levels of ADAM metallopeptidase domain 10 (ADAM10). The present study aimed to explore the role of ADAM10 in podocyte damage. Therefore, the expression of ADAM10 in high glucose (HG)-stimulated podocytes was measured by reverse transcription-qPCR and western blot. Moreover, the effects of ADAM10 knockdown on podocyte inflammation and apoptosis were determined by ELISA, western blot and TUNEL assay after confirming the efficacy of cell transfection. Subsequently, the effects of ADAM10 knockdown on the MAPK pathway and pyroptosis were assessed by western blot. Through performing the aforementioned experiments, the role of the MAPK pathway in the regulatory effects of ADAM10 was then investigated by pretreating podocytes with pathway agonists. ADAM10 expression was upregulated in HG-stimulated podocytes, while ADAM10 knockdown suppressed inflammation, apoptosis and pyroptosis of HG-stimulated podocytes and inhibited the activation of the MAPK signaling pathway. However, when podocytes were pretreated with pathway agonists (LM22B-10 or p79350), the aforementioned effects of ADAM10 knockdown were suppressed. The present study demonstrated that ADAM10 knockdown suppressed the inflammation, apoptosis and pyroptosis of HG-stimulated podocytes by blocking the MAPK signaling pathway.

TRIM40 ameliorates diabetic retinopathy through suppressing inflammation via Reelin/DAB1 signaling disruption: A mechanism by proteasomal degradation of DAB1

Diabetic retinopathy (DR) is a common microvascular complication of diabetes mellitus. Reelin, an extracellular matrix protein, and its effector protein Disabled1 (DAB1) have been linked to cellular events and retinal development. However, whether and how Reelin/DAB1 signaling causes DR remains to be investigated. In our study, significantly increased expression of Reelin, very low density lipoprotein receptor (VLDLR), ApoE receptor 2 (ApoER2) and phosphorylated DAB1 in retinas of streptozotocin (STZ)-induced DR mouse model was observed, along with enhanced expression of proinflammatory factors. Similar results are confirmed in high glucose (HG)-treated human retinal pigment epithelium cell line ARPE-19. Surprisingly, dysregulated tripartite motif-containing 40 (TRIM40), an E3 ubiquitin ligase, is found to be involved in DR progression by bioinformatic analysis. We observe a negative correlation between TRIM40 and p-DAB1 protein expression levels under HG conditions. Importantly, we find that TRIM40 over-expression markedly ameliorates HG-induced p-DAB1, PI3K, p-protein B kinase (AKT) and inflammatory response in HG-treated cells, but dose not affect Reelin expression. Of note, Co-IP and double immunofluorescence identify an interaction between TRIM40 and DAB1. Furthermore, we show that TRIM40 enhances K48-linked polyubiquitination of DAB1, thereby promoting DAB1 degradation. Finally, promoting TRIM40 expression by intravenous injection of the constructed adeno-associated virus (AAV-TRIM40) markedly ameliorates DR phenotypes in STZ-treated mice, as indicated by the decreased blood glucose and glycosylated hemoglobin (HbAlc) levels, and increased hemoglobin contents. Additionally, diabetes-related elevation of acellular capillaries was also meliorated in mice over-expressing TRIM40. The electroretinogram (ERG) deficits were strongly rescued in mice receiving AAV-TRIM40 injection. Moreover, AAV-TRIM40 attenuates the inflammation and p-DAB1 expression in retinal tissues of STZ-treated mice. Collectively, our findings disclose a mechanism through which TRIM40 limits DAB1 stability under physiological conditions and reveals TRIM40 as a potential therapeutic target for the intervention of Reelin/DAB1 signaling, contributing to DR treatment.

Serum metabolomic profiles and semaphorin-3A as biomarkers of diabetic retinopathy progression

Diabetic retinopathy (DR) is a typical microvascular complication of diabetes mellitus (DM) and it remains one of the leading causes of vision loss worldwide. Studies postulated that a distinct metabolic signature of DR exists and can be resolved from that of diabetes alone. Serum Semaphorin3A (Sema3A) levels have also been found to be correlated with the phenotypes of diabetic retinopathy. We aimed to analyze and identify serum metabolites and serum Sema3A levels that could be useful biomarkers of DR progression. This cross-sectional study included 45 type 2 diabetes (T2D) patients. Diabetic patients were divided into three groups based on the status of their complications: non-DR (NDR, n=15), non-proliferative DR (NPDR, n=15), and proliferative DR (PDR, n=15) groups. Serum metabolomic profiles of these patients were determined by using high-performance liquid chromatography-mass spectrometry (HPLC-MS), and serum Sema3A levels measured by ELISA. Metabolomics analysis revealed a set of metabolites that were altered in the serum of PDR patients as compared with NPDR and NDR groups. Among these metabolites total asymmetric dimethylarginine (ADMA) and Kynurenine were potential predictors of PDR patients. Significantly higher serum levels of Sema3A in PDR patients as compared with NPDR and NDR groups (p < 0.001), their serum levels were positively correlated with the central macular thickness (r= 0.952, p < 0.001) and negatively correlated with the superficial macular density (r=-0.952, p < 0.001). In conclusion, the metabolite signatures identified in this study and serum Sema3A levels could be proposed as biomarkers for DR development and progression in T2D patients. However, Sema3A was superior to metabolomics in the prediction of the severity of DR.

Sitagliptin Mitigates Diabetic Nephropathy in a Rat Model of Streptozotocin-Induced Type 2 Diabetes: Possible Role of PTP1B/JAK-STAT Pathway.

Diabetic nephropathy (DN) is a microvascular complication of diabetes mellitus. This study examined the therapeutic effects of sitagliptin, a dipeptidyl peptidase inhibitor, on DN and explored the underlying mechanism. Male Wistar albino rats (n = 12) were intraperitoneally administered a single dose of streptozotocin (30 mg/kg) to induce diabetes. Streptozotocin-treated and untreated rats (n = 12) were further divided into normal control, normal sitagliptin-treated control, diabetic control, and sitagliptin-treated diabetic groups (n = 6 in each). The normal and diabetic control groups received normal saline, whereas the sitagliptin-treated control and diabetic groups received sitagliptin (100 mg/kg, p.o.). We assessed the serum levels of DN and inflammatory biomarkers. Protein tyrosine phosphatase 1 B (PTP1B), phosphorylated Janus kinase 2 (P-JAK2), and phosphorylated signal transducer activator of transcription (P-STAT3) levels in kidney tissues were assessed using Western blotting, and kidney sections were examined histologically. Sitagliptin reduced DN and inflammatory biomarkers and the expression of PTP1B, p-JAK2, and p-STAT3 (p < 0.001) and improved streptozotocin-induced histological changes in the kidney. These results demonstrate that sitagliptin ameliorates inflammation by inhibiting DPP-4 and consequently modulating the PTP1B-related JAK/STAT axis, leading to the alleviation of DN.

Triptolide protects against podocyte injury in diabetic nephropathy by activating the Nrf2/HO-1 pathway and inhibiting the NLRP3 inflammasome pathway

Objectives Diabetic nephropathy (DN) is the most common microvascular complication of diabetes mellitus. This study investigated the mechanism of triptolide (TP) in podocyte injury in DN. Methods DN mouse models were established by feeding with a high-fat diet and injecting with streptozocin and MPC5 podocyte injury models were induced by high-glucose (HG), followed by TP treatment. Fasting blood glucose and renal function indicators, such as 24 h urine albumin (UAlb), serum creatinine (SCr), blood urea nitrogen (BUN), and kidney/body weight ratio of mice were examined. H&E and TUNEL staining were performed for evaluating pathological changes and apoptosis in renal tissue. The podocyte markers, reactive oxygen species (ROS), oxidative stress (OS), serum inflammatory cytokines, nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway-related proteins, and pyroptosis were detected by Western blotting and corresponding kits. MPC5 cell viability and pyroptosis were evaluated by MTT and Hoechst 33342/PI double-fluorescence staining. Nrf2 inhibitor ML385 was used to verify the regulation of TP on Nrf2. Results TP improved renal function and histopathological injury of DN mice, alleviated podocytes injury, reduced OS and ROS by activating the Nrf2/heme oxygenase-1 (HO-1) pathway, and weakened pyroptosis by inhibiting the nod-like receptor (NLR) family pyrin domain containing 3 (NLRP3) inflammasome pathway. In vitro experiments further verified the inhibition of TP on OS and pyroptosis by mediating the Nrf2/HO-1 and NLRP3 inflammasome pathways. Inhibition of Nrf2 reversed the protective effect of TP on MPC5 cells. Conclusions Overall, TP alleviated podocyte injury in DN by inhibiting OS and pyroptosis via Nrf2/ROS/NLRP3 axis.

Impact of vitamin B6 deficiency on the severity of diabetic peripheral neuropathy – A cross sectional study

BackgroundDiabetic Peripheral Neuropathy is one of the most important and significantly prevalent microvascular complications of Diabetes Mellitus. Pyridoxine is a key nutrient for protecting nerve health. The objective of this research is to study the prevalence rate of pyridoxine deficiency in Diabetic neuropathy patients, to understand the correlation between various biochemical and markers of diabetic neuropathy and pyridoxine deficiency. Results249 patients were selected for the study based on the selection criteria participants. 51.8% prevalence of pyridoxine deficiency in Diabetic neuropathy patients. The nerve conduction velocity significantly reduced in pyridoxine deficiency cases (p < 0.05). A strong inverse relationship is observed with fasting blood sugar levels and glycated hemoglobin pyridoxine deficiency might contribute to impaired glucose tolerance. ConclusionThere also exists a strong inverse relationship with glycemic markers. Significant direct correlation is observed with nerve conduction velocity. Pyridoxine also has properties of antioxidant which may be utilized for the management of Diabetic Neuropathy.

Efficacy of Huangkui Capsules in the Treatment of Diabetic Kidney Disease: A Systematic Review and Using Network Pharmacology

ABSTRACT Diabetic kidney disease (DKD), a severe and common microvascular complication of diabetes mellitus, has become a major cause of end-stage renal disease. Huangkui capsule (HKC) has been widely used to treat DKD. This meta-analysis aimed to provide high-quality evidence for the clinical application of HKC in DKD. The following databases: China National Knowledge Infrastructure, Wanfang Database, Chongqing VIP, SinoMed, Web of Science, EMBASE, PubMed, and The Cochrane Library, were searched for randomized controlled trials using the search theme: “angiotensin-converting enzyme inhibitor (ACEI)/angiotensin receptor blocker (ARB) combined with HKC for treatment of DKD” from their inception dates till August 2022. Studies were selected following our inclusion and exclusion criteria, and we extracted the required data. RevMan 5.3 was used for data statistics and analysis. Based on the main components identified by high performance liquid chromatography, the SwissADME, SwissTargetPrediction, and UniProt databases were used to predict the target genes of HKC. OMIM, DrugBank, GeneCards, and DisGeNet databases were used to predict DKD-related target genes. Venny 2.0 was then used to find the common targets in HKC and DKD. We conducted an HKC-ingredients-targets-DKD network using Cytoscape and a protein-protein interaction (PPI) network using the STRING database. Finally, we performed a Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis using the DAVID database. Exactly 15 studies were included in this meta-analysis. Compared with the control group using ACEI/ARB only, ACEI/ARB combined with HKC was found to significantly lower the urine albumin excretion rate (UAER; standard mean difference [SMD] = −1.92, 95% confidence interval [CI] [−2.93, −0.92]), 24h urinary total protein (24h-UTP; mean difference [MD] = −0.41, 95% CI [−0.64, −0.18], P = 0.0004), serum creatinine (SCr; SMD = −0.68, 95% CI [−1.02, −0.34]), blood urea nitrogen (BUN; SMD = −0.59, 95% CI [−1.04, 0.14], P = 0.01), total cholesterol (TC; MD = −1.22, 95% CI [−2.04, 0.39], P = 0.004, and triglyceride (TG) levels (MD = −0.54, 95% CI [−0.94, −0.15], P = 0.007). There was no significant difference in low-density lipoprotein cholesterol levels (MD = −0.45, 95% CI [−1.21, 0.31], P = 0.24) and adverse effects (RR = 0.98, 95% CI [0.43, 2.22], P = 0.96) between the two groups. Exactly 127 genes were obtained using network pharmacology and were the common target genes of HKC and DKD. PPI network showed that the key targets are SRC, AKT1, HSP90AA1, PIK3R1, SYK, FYN, ESR1, and F2. GO analysis and KEGG pathway enrichment analysis showed that HKC could alleviate the pathological glomerular changes by inhibiting the activity of the PI3K/Akt signaling pathway. The combination of ACEI/ARB and HKC has a good safety profile and may be effective for the prevention of DKD by reducing UAER, 24h-UTP, TC, and TG levels. Furthermore, HKC may treat DKD primarily by inhibiting the PI3K-Akt signaling pathway, and further experimental studies are required to verify this hypothesis.

Astragalin ameliorates renal injury in diabetic mice by modulating mitochondrial quality control via AMPK-dependent PGC1α pathway.

Diabetic kidney disease (DKD) is a common microvascular complication of diabetes mellitus, and oxidative stress and mitochondrial dysfunction play an important role in this process. It has been shown that aldose reductase (ALR2) catalyzes NADPH-dependent reduction of glucose to sorbitol, resulting in oxidative stress and mitochondrial dysfunction in diabetic patients. Astragalin (AG), a flavonoid extracted from Thesium chinense Turcz., shows an inhibitory activity on ALR2. In this study, we investigated the therapeutic effects of AG against renal injury in streptozocin (STZ)-induced diabetic mouse model. Diabetic mice were orally administered AG (5, 10 mg·kg-1·d-1) for 4 weeks. We showed that AG treatment greatly improved the proteinuria and ameliorated renal pathological damage without affecting the elevated blood glucose in diabetic mice. Furthermore, AG treatment significantly suppressed highly activated ALR2, and reduced oxidative stress in the kidney of diabetic mice and in high glucose and lipids-stimulated HK2 cells in vitro. We demonstrated that AG treatment modulated mitochondrial quality control and ameliorated apoptosis, boosting mitochondrial biogenesis, maintaining mitochondrial dynamic homeostasis, and improving energy metabolism disorder in vivo and in vitro. In high glucose and lipids-stimulated HK2 cells, we found that AG (20 μM) restored the phosphorylation level of AMPK, and upregulated the expression and transcriptional activity of PGC1α, whereas treatment with H2O2, blockade of AMPK with Compound C or knockdown of AMPKα with siRNA abolished the protective effect of AG on mitochondrial function, suggesting that antioxidant effects and activation of AMPK-dependent PGC1α pathway might be the molecular mechanisms underlying the protective effects of AG on mitochondrial quality control. We conclude that AG could be a promising drug candidate for the treatment of diabetic renal injury through activating AMPK.

TFEB attenuates hyperglycemia-induced retinal capillary endothelial cells injury via autophagy regulation.

Diabetic retinopathy is a common microvascular complication of diabetes mellitus. The maintenance of retinal capillary endothelial cell homeostasis requires a complete and unobtrusive flow of autophagy because it may help combat the inflammatory response, apoptosis, and oxidative stress damage of cells in diabetes mellitus. The transcription factor EB is a master regulator of autophagy and lysosomal biogenesis, but its role in diabetic retinopathy remains unknown. This study aimed to confirm the involvement of transcription factor EB in diabetic retinopathy and explore the role of transcription factor EB in hyperglycemia-linked endothelial injury in vitro. First, the expression levels, including the nuclear location of transcription factor EB and autophagy, were reduced in diabetic retinal tissues and high glucose-treated human retinal capillary endothelial cells. Subsequently, autophagy was mediated by transcription factor EB in vitro. Moreover, transcription factor EB overexpression reversed high glucose-induced autophagy inhibition and lysosomal dysfunction and protected human retinal capillary endothelial cells from inflammation, apoptosis, and oxidative stress damage caused by high glucose treatment. Additionally, under high-glucose stimulation, the autophagy inhibitor chloroquine attenuated transcription factor EB overexpression-mediated protection, and the autophagy agonist Torin1 rescued transcription factor EB knockdown-induced damage effects. Taken together, these results suggest that transcription factor EB is involved in the development of diabetic retinopathy. In addition, transcription factor EB protects human retinal capillary endothelial cells from high glucose-induced endothelial damage via autophagy.

Patient Characteristics and Prevalence of Single Nucleotide Polymorphism (SNP) V16A MnSOD Gene in Diabetic Retinopathy Patients at RSUP Prof. Dr. I.G.N.G. Ngoerah Denpasar Bali

Diabetic Retinopathy (DR) is a microvascular complication of Diabetes Mellitus (DM) caused by hyperglycemia and oxidative stress that is prevalent to the blindness risk. Manganese superoxide dismutase (MnSOD) gene affect the oxidative damage and progression of DR severity. This study aims to determine the association of patient characteristics on the severity and prevalence of Single Nucleotide Polymorphism (SNP) V16A in the MnSOD gene in DR patients at Prof. Dr. I.G.N.G. Ngoerah Denpasar Bali. Twelve (12) subjects with Non-Proliferative DR and 21 subjects with Proliferative DR were enrolled. Patient characteristics (age, T2DM duration, BMI, blood pressure) were associated to the DR severity using cross-sectional analytic study. While to detect the status of the SNP V16A MnSOD gene using descriptive experimental study. DNA segment amplification was performed using the PCR method, sequencing was performed to detect the base pattern at the location of the SNP. A domination of the VV genotype in the SNP V16A MnSOD gene was present in all subjects (100%). The majority of subjects were female (51,5%), age range 40-65 years (84,9%), T2DM duration &lt;10 years (60,6%), normal BMI (60,6%). Blood pressure was dominated with non hypertension 60,6%, with a history of oral anti hypertensive. No significant association was found between patient characteristics and the DR severity (p&lt;0,05). We may conclude that wildtype variant (VV) SNP V16A MnSOD gene was dominated in this finding. There is no significant association between patient characteristics and the severity of DR in Prof. Dr. I.G.N.G. Ngoerah Denpasar Bali. Keywords: Diabetic Retinopathy, MnSOD gene, SNP V16A

Convolutional Neural Networks Accurately Identify Ungradable Images in a Diabetic Retinopathy Telemedicine Screening Program.

Purpose: Diabetic retinopathy (DR) is a microvascular complication of diabetes mellitus (DM). Standard of care for patients with DM is an annual eye examination or retinal imaging to assess for DR, the latter of which may be completed through telemedicine approaches. One significant issue is poor-quality images that prevent adequate screening and are thus ungradable. We used artificial intelligence to enable point-of-care (at time of imaging) identification of ungradable images in a DR screening program. Methods: Nonmydriatic retinal images were gathered from patients with DM imaged during a primary care or endocrinology visit from September 1, 2017, to June 1, 2021. The Topcon TRC-NW400 retinal camera (Topcon Corp., Tokyo, Japan) was used. Images were interpreted by 5 ophthalmologists for gradeability, presence and stage of DR, and presence of non-DR pathologies. A convolutional neural network with Inception V3 network architecture was trained to assess image gradeability. Images were divided into training and test sets, and 10-fold cross-validation was performed. Results: A total of 1,377 images from 537 patients (56.1% female, median age 58) were analyzed. Ophthalmologists classified 25.9% of images as ungradable. Of gradable images, 18.6% had DR of varying degrees and 26.5% had non-DR pathology. 10 fold cross-validation produced an average area under receiver operating characteristic curve (AUC) of 0.922 (standard deviation: 0.027, range: 0.882 to 0.961). The final model exhibited similar test set performance with an AUC of 0.924. Conclusions: This model accurately assesses gradeability of nonmydriatic retinal images. It could be used for increasing the efficiency of DR screening programs by enabling point-of-care identification of poor-quality images.

Association of C-peptide level with peripheral neuropathy in type 2 diabetes: An observational cross-sectional preliminary study

Backround and aimsPeripheral neuropathy is the most common microvascular complication of diabetes mellitus. In subjects with type 1 diabetes (T1D) relationship of C-peptide levels and neuropathy has been observed in several studies, however, there are very few studies in type 2 diabetes (T2D) subjects. In this study we aim to assess the association of C-peptide levels with peripheral neuropathy in Indian subjects with T2D. MethodOne hundred patients of T2D were included in this study. Clinical and laboratory parameter was assessed for all participants. The C-peptide level was measured by fluorometric enzyme immunoassay method. Assessment of diabetic peripheral neuropathy was based on diabetic neuropathy symptom score and the diabetic neuropathy examination scores. ResultsTotal 100 patients completed the study. Mean age of subjects was 60.03 years and male: female ratio was 1.17. Peripheral neuropathy was detected in 47% of subjects evaluated. Subjects were further divided in to neuropathy group and no-neuropathy group for analysis. Age in neuropathy group was significantly higher than no-neuropathy group [65.62 ± 10.5 vs 55.08 ± 9.41 yrs (p-value <0.0001)] and similarly duration of T2D was significantly higher in neuropathy group [10.11 ± 6.13 vs 4.16 ± 3.7 yrs (p-value <0.0001)]. Importantly mean fasting C-peptide (2.27 ± 0.98 vs 3.12 ± 0.84 ng/ml) and mean post meal C-peptide (4.27 ± 1.34 vs 5.33 ± 0.89 ng/ml) were significantly lower in neuropathy group compared to no-neuropathy group. An association of HbA1c level and neuropathy was statistically not significant (p = 0.793). ConclusionSerum C-peptide concentrations are associated with peripheral neuropathy in T2DM patients, independent of the degree of glycemic control.

Transplantation of Human Amniotic Mesenchymal Stem Cells Up-Regulates Angiogenic Factor Expression to Attenuate Diabetic Kidney Disease in Rats.

Diabetic kidney disease (DKD) is a prevalent and intractable microvascular complication of diabetes mellitus (DM), the process of which is closely related to abnormal expression of angiogenesis-regulating factors (ARFs). Stem cell transplantation might be a novel strategy for treating DKD. This study aims to explore the effect of transplantation of human amniotic mesenchymal stem cells (hAMSCs) on renal microangiopathy in a type 1 DKD rat model (T1DRM). Seventy-two rats were randomly divided into three groups, including normal control group, DKD group, and hAMSCs transplantation group. T1DRM was established using a rat tail vein injection of streptozotocin (STZ) (55 mg/kg). hAMSCs were obtained from placental amniotic membranes during cesarean delivery and transplanted at 3 and 4 weeks through penile veins. At 6, 8, and 12 weeks following transplantation, blood glucose levels, renal function, pathological kidney alterations, and the expressions of ARFs' mRNA and protein were analyzed. In T1DRM, transplanted hAMSCs that were homed at the injured site of kidneys increased ARFs' expression and decreased blood glucose levels. Compared to the DKD group, the levels of 24-h urinary protein, serum creatinine, urea, and kidney injury molecule-1 (KIM-1) were reduced in hAMSCs transplantation group. In terms of renal pathology such as the degree of basement membrane thickening, hAMSCs transplantation was also less severe than the DKD group, thereby alleviating kidneyinjury. hAMSCs transplantation might ameliorate STZ-induced chronic kidney injury through increasing ARFs' expression in kidneys and lowering blood glucose levels.

The effect of crocin and losartan on TGF-β gene expression and histopathology of kidney tissue in a rat model of diabetic nephropathy.

Diabetic nephropathy is one of the most common microvascular complications of diabetes mellitus that finally leads to complete loss of kidney function. Therefore, this study aimed to evaluate the effect of crocin and losartan on TGF-β gene expression and histopathology of kidney tissue in a rat model of diabetic nephropathy. Forty male Wistar rats were randomly divided into five groups (n=8): Untreated control, Diabetic (D), D + crocin, D + losartan, and D + losartan + crocin. Induction of diabetes was performed using streptozotocin (50 mg/kg/ Intraperitoneal injection). At the end of the eight-week period, the rats were sacrificed. Spectrophotometry measured serum glucose, urea, creatinine, and uric acid levels. Microalbumin and creatinine levels were measured in 24-hour urine. Real-time PCR was used to determine the relative expression of the TGF-β gene in kidney tissue. Renal tissue histopathology was also examined. The results showed that hyperglycemia increased biochemical factors associated with diabetes, TGF-β gene expression, and kidney damage. Separate treatment with crocin and losartan led to a decrease in renal function factors and TGF-β gene expression and improved kidney damage. Our results showed that crocin could improve kidney function in diabetic conditions. In addition, we showed that crocin increases the effectiveness of losartan. Consequently, we suggest that crocin in combination with chemical drugs can be a potential therapeutic agent for diabetes and its complications. Nonetheless, human studies are needed to make firm findings.