High uric acid aggravates apoptosis of lung epithelial cells induced by cigarette smoke extract through downregulating PRDX2 in chronic obstructive pulmonary disease.

Cigarette smoke exposure is the major cause of chronic obstructive pulmonary disease (COPD). Cigarette smoke heightens the elevation of reactive oxygen species (ROS) and thus leads to apoptosis. Hyperuricemia has been considered as a risk factor for COPD. However, the underlying mechanism for this aggravating effect remains unclear. The current study sought to examine the role of high uric acid (HUA) in COPD using cigarette smoke extract (CSE) exposed murine lung epithelial (MLE-12) cells. Our data showed that CSE induced the increase of ROS, mitochondrial dynamics disorder, and apoptosis, while HUA treatment aggravated the effects of CSE. Further studies suggested that HUA decreased the expression of antioxidant enzyme-peroxiredoxin-2 (PRDX2). Overexpression of PRDX2 inhibited excessive ROS generation, mitochondrial dynamics disorder, and apoptosis induced by HUA. Knockdown of PRDX2 by small interfering RNA (siRNA) promoted ROS generation, mitochondrial dynamics disorder, and apoptosis in MLE-12 cells treated with HUA. However, antioxidant N-acetylcysteine (NAC) reversed the effects of PRDX2-siRNA on MLE-12 cells. In conclusion, HUA aggravated CSE-induced cellular ROS levels and led to ROS-dependent mitochondrial dynamics disorder and apoptosis in MLE-12 cells through downregulating PRDX2.

Pentoxifylline decreases the activity of the nucleotide-binding oligomerization domain-like receptor protein 3 pathway: potential role for preventing arteriovenous fistula stenosis.

PURPOSE: This study aimed to determine the effect of pentoxifylline (PTX) on the nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome pathway and its role in preventing arteriovenous fistula (AVF) failure. METHODS: Vein samples were collected from AVF failure patients and from patients who underwent surgical AVF as a control. The expressions of CD34 and NLRP3 in AVF tissues were detected by immunohistochemistry and Western blotting. Arteriovenous fistula rat models were established by the end-to-end anastomosis of the common carotid artery and external jugular vein. The AVF models were divided into the following groups: AVF, AVF + PTX, AVF + uraemia and AVF + uraemia + PTX. Six weeks after surgery, the AVF tissues in each group were collected to detect the expressions of CD34, NLRP3, caspase-1 and interleukin (IL)-1ß by immunohistochemistry, Western blotting and real-time polymerase chain reaction. RESULTS: The expressions of NLRP3 and CD34 in human AVF failure tissues were significantly higher than those in normal veins (p < 0.001), indicating that NLRP3 was upregulated in patients with AVF failure. In our animal study, the veins in the AVF + uraemia group exhibited heavy hyperplasia, and the boundary between the media and the adventitia was not clear. However, PTX alleviated this hyperplasia. Compared with the AVF models, the AVF + uraemia models had much higher expressions of NLRP3, caspase-1, IL-1ß and CD34 (p < 0.001). However, PTX had the opposite effect against uraemia on the NLRP3 inflammasome pathway at both the gene and protein levels. CONCLUSIONS: Our findings provide new insights that show that PTX can decrease the activity of the NLRP3 inflammasome pathway in AVF models. Pentoxifylline has the potential as a drug for preventing intimal hyperplasia and AVF failure.

Association of Thyroid Hormone Levels with Microvascular Complications in Euthyroid Type 2 Diabetes Mellitus Patients.

Purpose: To examine the prevalence of different microvascular complications and investigate the association between thyroid hormones (THs) and these complications in euthyroid patients with type 2 diabetes mellitus (T2DM). Methods: A total of 248 T2DM patients were analyzed retrospectively for the study. All patients received a detailed and standard assessment to identify diabetic peripheral neuropathy (DPN), diabetic nephropathy (DN), and diabetic retinopathy (DR). Multivariate logistic regression was carried out to analyze the association between THs and diabetic microvascular complications. Results: The study found the prevalence of any microangiopathy to be 72.18% (n = 179). At the same time, the prevalence of DPN was 54.84% (n=136), while that of DN was 31.85% (n=79). Likewise, the prevalence of DR was 35.48% (n=88). The odds ratios (ORs) for free triiodothyronine (FT3) developing any microangiopathy, DPN, DN and DR were 0.200, 0.361, 0.310, and 0.588 (P<0.05), respectively. Also, the ORs for free thyroxine (FT4) developing any microangiopathy, DPN, DN and DR were 0.643, 0.800, 0.702 and 0.726 (P<0.05), respectively. Lastly, the ORs for thyroid-stimulating hormone (TSH) developing DPN was 1.57 (95% CI: 1.148-2.137, P=0.005). Conclusion: The study concludes that serum FT3 and FT4 levels are negatively associated with any microangiopathy, DPN, DN and DR in euthyroid patients with T2DM, independent of traditional risk factors. However, the TSH levels are positively associated with DPN. Future larger sample-size studies are needed to confirm the relationship between thyroid hormone levels and microvascular complications in euthyroid patients with T2DM.

Astragaloside IV Alleviates Renal Tubular Epithelial-Mesenchymal Transition via CX3CL1-RAF/MEK/ERK Signaling Pathway in Diabetic Kidney Disease.

Background: Epithelial-mesenchymal transition (EMT) plays an important role in interstitial matrix deposition and renal fibrosis in diabetic kidney disease (DKD). It has been verified that Astragaloside IV (AS-IV) is beneficial for ameliorating DKD. However, the underlying mechanisms of AS-IV on regulating EMT in DKD are yet to be established. Accumulated evidence has suggested that C-X3-C motif ligand 1 (CX3CL1) plays a significant role in the progression of EMT. Purpose: We aimed to investigate whether AS-IV could alleviate EMT by regulating CX3CL1 in DKD and reveal its underlying mechanisms. Methods: For the in vivo study, mice were divided into the following five groups (n=10): db/m+vehicle, db/db+vehicle, db/db+AS-IV-L (10mg/kg/d), db/db+AS-IV-M (20mg/kg/d), db/db+AS-IV-H (40mg/kg/d). After 12 weeks of treatment, the renal injuries were assessed based on the related parameters of urine, blood and histopathological examination. Immunohistochemistry and Western blotting were used to detect relative proteins levels. Then in HK-2 cells, the molecular mechanism of AS-IV attenuating the EMT in mice with DKD through the CX3CL1-RAF/MEK/ERK pathway was studied. Results: In the present study, we found that AS-IV reduced urinary protein levels and improved renal pathological damage in DKD mice. Moreover, AS-IV ameliorated the renal tubular EMT induced by hyperglycemia or high glucose (HG), and decreased the expression of CX3CL1 and inhibited the activation of the RAF/MEK/ERK pathway in vivo and in vitro. In HK-2 cells, downregulation of CX3CL1 suppressed the stimulation of the RAF/MEK/ERK pathway and EMT induced by HG. However, CX3CL1 overexpression eliminated the benefits of AS-IV on the RAF/MEK/ERK pathway and EMT. Conclusion: In summary, we indicated that AS-IV alleviates renal tubular EMT through the CX3CL1-RAF/MEK/ERK signaling pathway, indicating that CX3CL1 could be a potential therapeutic target of AS-IV in DKD.

Associations of Insomnia With Hypertension and Coronary Artery Disease Among Patients With Type 2 Diabetes Mellitus.

Purpose: This study aimed to determine whether insomnia is associated with hypertension (HBP) and coronary artery disease (CAD) in a hospital-based sample of patients with type 2 diabetes mellitus (T2DM). Methods: Our present study included 354 patients with T2DM. According to the diagnostic criteria of insomnia, the participants were assigned to three groups based on the duration of T2DM and insomnia diagnosis. Patients with T2DM alone were placed in group A; patients with T2DM longer than insomnia were placed in group B; and patients with insomnia longer than T2DM were placed in group C. Medical history was collected from all the patients in detail. Besides, the participants underwent thorough physical examinations and laboratory measurements. Propensity score matching (PSM) was applied to evaluate the associations of insomnia with HBP and CAD. The univariate and multivariate logistic regression analysis was used to explore whether insomnia was a risk factor for HBP and CAD in patients with T2DM. Results: Of 354 patients, 225 patients were included in group A, 62 patients were included in group B, and 67 patients were included in group C. Compared with groups B and C, group A showed a lower prevalence of HBP and CAD (p < 0.05). In addition, compared with group B, group C showed no difference in the prevalence of HBP and CAD (p > 0.05). After PSM was performed, groups B and C had a higher prevalence of HBP and CAD (p < 0.05) than group A with no significant difference between groups B and C (p > 0.05). In the univariate and multivariate logistic regression analysis, insomnia was a risk factor for HBP [univariate: odds ratio (OR) = 3.376, 95% CI 2.290-6.093, p < 0.001; multivariate: OR = 2.832, 95% CI 1.373-5.841, p = 0.005] and CAD (univariate: OR = 5.019, 95% CI 3.148-8.001, p < 0.001; multivariate: OR = 5.289, 95% CI 2.579-10.850, p < 0.001). Conclusion: T2DM combined with insomnia was related to HBP and CAD and insomnia was a risk factor for HBP and CAD in patients with T2DM. However, larger, prospective studies are required to confirm our findings.

Functional Status of Pancreatic α and ß Cells in Type 2 Diabetes Mellitus Patients with Different Plasma Triglyceride Levels: A Retrospective Analysis.

OBJECTIVE: To investigate the functional status of pancreatic α and ß cells in Type 2 diabetes mellitus (T2DM) patients with different plasma triglyceride (TG) levels. TG levels can be prognostic markers for T2DM. METHODS: A total of 328 patients with T2DM were divided into three groups according to different TG levels: the TGL group: TG < 1.7 mmol/L; TGM group: 1.7 mmol/L ≤ TG < 2.3 mmol/L; and TGH group: TG ≥ 2.3 mmol/L. An oral glucose tolerance test (OGTT), insulin release test, and glucagon release test were performed in each patient. The changes of glucagon, glucagon/insulin ratio, early insulin secretion index (ΔI 30/ΔG 30), and area under the insulin curve (AUCI) were compared among each group. Also, the correlations between glucagon and pancreatic ß-cell function, glycosylated hemoglobin (HbA1c), and other indices were analyzed. RESULTS: With the increase of TG, the fasting and postprandial glucagon levels, the glucagon/insulin ratio, and the area under the glucagon curve (AUCG) presented an increasing trend. The homeostasis model assessment of insulin resistance (HOMA-IR) of the TGH group was significantly increased compared to the TGL and TGM groups. In addition to the increase in TG levels, the insulin sensitivity index (ISI), homeostasis model assessment for ß-cell function index (HOMA-ß), ΔI 30/ΔG 30, and AUCI displayed a reducing trend. Glucagon was negatively correlated with ΔI 30/ΔG 30, high-density lipoprotein (HDL), HOMA-ß, body mass index (BMI), ISI, and AUCI (P < 0.05) and positively correlated with fasting blood glucose (FPG), AUCG, HOMA-IR, HbA1c, duration, TG, low-density lipoprotein (LDL), and total cholesterol (TC) (P < 0.05). CONCLUSION: Hypertriglyceridemia aggravated the dysfunction of pancreatic α and ß cells. A reasonable control of the TG level makes it easier for blood glucose to reach the standard.

Complement C7 is Specifically Expressed in Mesangial Cells and is a Potential Diagnostic Biomarker for Diabetic Nephropathy and is Regulated by miR-494-3p and miR-574-5p.

BACKGROUND: Diabetic nephropathy (DN) is the leading cause of end-stage renal disease, but it remains relatively underdiagnosed. OBJECTIVE: In this study, we aimed to explore the key regulatory pathways and potential biomarkers related to DN using integrated bioinformatics analysis and validation. METHODS: First, the microarray data of the GSE30528 and GSE96804 datasets were downloaded from the Gene Expression Omnibus (GEO) database, and differentially expressed genes (DEGs) were screened. Then, weighted gene coexpression network analysis (WGCNA), gene ontology (GO) annotation, gene set enrichment analysis (GSEA) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to identify key pathways and genes. qRT-PCR and receiver operating characteristic (ROC) curves were used to validate our results. Furthermore, single-cell RNA sequencing (scRNA-seq) data were reanalyzed to investigate the expression specificity of C7 in DN cells. An online database search and luciferase reporter assay identified the target relationship between miRNAs and C7. RESULTS: The "complement and coagulation cascades" were significantly enriched, and complement C3 and C7 were candidate markers. The receiver operating characteristic (ROC) curve revealed that C7 had significant diagnostic value (AUC=0.865) in DN. Through scRNA-seq reanalysis, we found that C7 was specifically elevated in mesangial (MES) cells of DN. Moreover, we found that the expression of C7 was regulated by miR-494-3p and miR-574-5p. CONCLUSION: This is the first study to reveal that C7 is specifically expressed in mesangial cells, is a potential diagnostic biomarker for diabetic nephropathy, and is regulated by miR-494-3p and miR-574-5p.

Angelica Dahurica Regulated the Polarization of Macrophages and Accelerated Wound Healing in Diabetes: A Network Pharmacology Study and In Vivo Experimental Validation.

Diabetic wounds exhibit retarded and partial healing processes. Therefore, patients are exposed to an elevated risk of infection. It has been verified that Angelica dahurica (Hoffm.) Benth. and Hook. f. ex Franch. and Sav (A. dahurica) is conducive for wound healing. However, the pharmacological mechanisms of A. dahurica are yet to be established. The present study uses network pharmacology and in vivo experimental validation to investigate the underlying process that makes A. dahurica conducive for faster wound healing in diabetes patients. 54 potential targets in A. dahurica that act on wound healing were identified through network pharmacology assays, such as signal transducer and activator of transcription 3 (STAT3), JUN, interleukin-1ß (IL-1ß), tumor necrosis factor (TNF), and prostaglandin G/H synthase 2 (PTGS2). Furthermore, in vivo validation showed that A. dahurica accelerated wound healing through anti-inflammatory effects. More specifically, it regulates the polarization of M1 and M2 subtypes of macrophages. A. dahurica exerted a curative effect on diabetic wound healing by regulating the inflammation. Hence, pharmacologic network analysis combined with in vivo validation elucidated the probable effects and underlying mechanisms of A. dahurica's therapeutic effect on diabetic wound healing.

Associations of Thyroid Hormone Levels and Macrovascular Complications in Euthyroid Type 2 Diabetic Patients.

PURPOSE: The purpose of this study is to evaluate whether thyroid hormone in euthyroid patients with type 2 diabetes mellitus (T2DM) is associated with macrovascular complications. PATIENTS AND METHODS: The authors examined 311 patients enrolled from February 2019 to December 2019 in Tianjin Medical University Chu Hsien-I Memorial Hospital. A medical record review enabled the collection of demographic and anthropometric information. We classified the patients into two groups based on the echocardiography and vascular ultrasonography results, namely, non-macrovascular complications (n=131) group and macrovascular complications (n=180) group. Odds ratios (OR) and 95% confidence intervals (CI) were calculated, adjusting for potential confounders, the prevalence of macrovascular complications was determined using multivariate logistic regression. RESULTS: A significant association was observed for diabetic macrovascular complications with normal free triiodothyronine (FT3) (OR=0.534, 95% CI 0.358-0.796, p = 0.002) and free thyroxine (FT4) (OR= 0.844, 95% CI 0.760-0.937, p = 0.001). Nevertheless, there was no evidence of any association between thyroid-stimulating hormone (TSH) and the development of diabetic macrovascular complications. When stratified by the body mass index (BMI), a similar relationship existed with the overall results. The positive association remained in restricted analyses involving only patients with HbA1c abnormalities. CONCLUSION: Overweight or obese T2DM patients are at high risk due to the implicit association between low but clinically normal thyroid hormone levels and elevated risk of macrovascular complications. However, there were no statistically significant associations between TSH and diabetic macrovascular complications.

The ratio of estimated average glucose to fasting plasma glucose level as an indicator of insulin resistance in young adult diabetes: An observational study.

At present, glycated hemoglobin (HbA1c) and glycated albumin (GA) are used to evaluate glycemic control in diabetic patients, but they cannot reflect insulin deficiency and/or insulin resistance.We investigated the feasibility of using estimated average glucose to fasting plasma glucose ratio (eAG/fPG ratio) to estimate insulin resistance in young adult diabetes. A total of 387 patients with type 2 diabetes were included and were stratified into 2 groups based on median values of the glycemic index ratio: the GA/A1c ratio <2.09 (n = 91) and ≥2.09 (n = 296); the eAG/fPG ratio <1.69 (n = 155) and ≥1.69 (n = 232). HbA1c, GA, fructosamine, insulin, and C-peptide levels were measured. The ratio of GA to HbA1c was calculated, and the homeostasis model assessment of ß-cell function and insulin resistance were determined. The homeostasis model assessment of insulin resistance level was significantly associated with the eAG/fPG ratio, but not with the ratio of GA to HbA1c, GA, HbA1c, and fructosamine levels. The ratio of estimated average glucose to fasting plasma glucose level correlates with insulin resistance in young adult diabetes.

Angelica dahurica promoted angiogenesis and accelerated wound healing in db/db mice via the HIF-1α/PDGF-ß signaling pathway.

Impaired angiogenesis is crucial for impeding the wound healing process in diabetic foot ulcers (DFUs). In this study, we found that Angelica dahurica (A. dahurica) stimulated angiogenesis and benefited wound healing in genetic mouse models of diabetes. In HUVECs, A. dahurica promoted cell proliferation and tube formation, which was accompanied by increased nuclear translocation of HIF-1α under hypoxic conditions and led to elevated PDGF-ß protein expression. A. dahurica activated the PI3K/AKT signaling pathway in human umbilical vein endothelial cells (HUVECs), which was abrogated by the PI3K inhibitor LY294002. Furthermore, the cellular expression of PDGF-ß decreased significantly after treatment with a HIF-1α-siRNA, and PDGF-ß expression was increased in HIF-1α-overexpressing cells. In a full-thickness cutaneous wound healing db/db mouse model, A. dahurica accelerated wound closure, which was reflected by a significantly reduced wound area and an increase in neovascularization, as well as by elevated PDGF-ß expression and increased capillary formation. In addition, A. dahurica activated the PI3K/AKT signaling pathway and enhanced HIF-1α synthesis in wounds. In summary, A. dahurica promoted angiogenesis of HUVECs in vitro by promoting signaling via the HIF-1α/PDGF-ß pathway, efficiently enhancing vascularization in regenerated tissue and facilitating wound healing in vivo. The results revealed that A. dahurica has potential as a therapy for vessel injury-related wounds.

BML-111 attenuates high glucose-induced inflammation, oxidative stress and reduces extracellular matrix accumulation via targeting Nrf2 in rat glomerular mesangial cells.

Diabetic nephropathy (DN) is the most paradigmatic complication of diabetes mellitus (DM) and brings about severe social and economic burdens. BML-111 is a potent agonist of Lipoxin A4 and has shown anti-inflammatory function in many diseases. The aim of the study is to investigate the effects of BML-111 on high glucose (HG) -induced mesangial cells. HBZY-1 cells were stimulated by HG with or without BML-111. ML385 was used as an Nrf2 inhibitor. Cell proliferation was measured by CC-K 8 assay. Besides, levels of TNF-α, IL-1, IL-6 and MCP-1 were detected by corresponding ELISA kits. DCFH-DA staining and an available ROS kit were employed to determine the ROS generation. In addition, extracellular matrix (ECM) accumulation was evaluated by immunofluorescence assay and western blot analysis. The protein expressions involved in Nrf2/HO-1 and MAPK pathway were assessed by western blot assay. Results indicated that BML-111 extremely inhibited HBZY-1 cell proliferation induced by HG. Moreover, BML-111 reduced the levels of TNF-α, IL-1, IL-6 and MCP-1, declined intracellular ROS level, and attenuated expression of ECM proteins laminin, fibronectin, collagen IV and TGF-ß1. In addition, BML-111 promoted the activation of Nrf2, HO-1, and NQO1, while suppressed the phosphorylation of p38 and JNK. Further, NRF2 silence reversed the inhibitory effects of BML-111 on HG-induce inflammation, oxidative stress and ECM accumulation, accelerate the MAPK signaling, and diminished the expression of Nrf2 pathway. In summary, BML-111 alleviated HG-induced injury in HBZY-1 cells by repressing inflammatory response, oxidative stress and ECM accumulation via activating Nrf2 and inhibiting MAPK pathway.

Angelica Dahurica ethanolic extract improves impaired wound healing by activating angiogenesis in diabetes.

Abnormal angiogenesis plays an important role in impaired wound healing and development of chronic wounds in diabetes mellitus. Angelica dahurica radix is a common traditional Chinese medicine with wide spectrum medicinal effects. In this study, we analyzed the potential roles of Angelica dahurica ethanolic extract (ADEE) in correcting impaired angiogenesis and delayed wound healing in diabetes by using streptozotocin-induced diabetic rats. ADEE treatment accelerated diabetic wound healing through inducing angiogenesis and granulation tissue formation. The angiogenic property of ADEE was subsequently verified ex vivo using aortic ring assays. Furthermore, we investigated the in vitro angiogenic activity of ADEE and its underlying mechanisms using human umbilical vein endothelial cells. ADEE treatment induced HUVECs proliferation, migration, and tube formation, which are typical phenomena of angiogenesis, in dose-dependent manners. These effects were associated with activation of angiogenic signal modulators, including extracellular signal-regulated kinase 1/2 (ERK1/2), Akt, endothelial nitric oxide synthase (eNOS) as well as increased NO production, and independent of affecting VEGF expression. ADEE-induced angiogenic events were inhibited by the MEK inhibitor PD98059, the PI3K inhibitor Wortmannin, and the eNOS inhibitor L-NAME. Our findings highlight an angiogenic role of ADEE and its ability to protect against impaired wound healing, which may be developed as a promising therapy for impaired angiogenesis and delayed wound healing in diabetes.

The Four-Herb Chinese Medicine Formula Tuo-Li-Xiao-Du-San Accelerates Cutaneous Wound Healing in Streptozotocin-Induced Diabetic Rats through Reducing Inflammation and Increasing Angiogenesis.

Impaired wound healing in diabetic patients is a serious complication that often leads to amputation or even death with limited effective treatments. Tuo-Li-Xiao-Du-San (TLXDS), a traditional Chinese medicine formula for refractory wounds, has been prescribed for nearly 400 years in China and shows good efficacy in promoting healing. In this study, we explored the effect of TLXDS on healing of diabetic wounds and investigated underlying mechanisms. Four weeks after intravenous injection of streptozotocin, two full-thickness excisional wounds were created with a 10 mm diameter sterile biopsy punch on the back of rats. The ethanol extract of TLXDS was given once daily by oral gavage. Wound area, histological change, inflammation, angiogenesis, and collagen synthesis were evaluated. TLXDS treatment significantly accelerated healing of diabetic rats and improved the healing quality. These effects were associated with reduced neutrophil infiltration and macrophage accumulation, enhanced angiogenesis, and increased collagen deposition. This study shows that TLXDS improves diabetes-impaired wound healing.