Efficacy and safety of empagliflozin in combination with insulin in Chinese patients with type 2 diabetes and insufficient glycaemic control: A phase III, randomized, double-blind, placebo-controlled, parallel study.

AIM: To evaluate the efficacy and safety of empagliflozin in combination with insulin ± oral antidiabetic drugs (OADs) over 24 weeks, in Chinese patients with type 2 diabetes (T2D) who had insufficient glycaemic control. MATERIALS AND METHODS: This was a randomized, double-blind, placebo-controlled, parallel group, multicentre phase III study. Adult patients with T2D and insufficient glycaemic control who received insulin ± up to two OADs were randomized (1:1:1) to receive empagliflozin 10 or 25 mg, or placebo for 24 weeks. The primary endpoint was change from baseline in HbA1c at week 24. RESULTS: Of 219 randomized patients, 73 patients were in each treatment group; baseline characteristics were comparable among the groups. There was a significantly larger decrease from baseline in HbA1c (adjusted mean treatment difference -0.99 and -0.98 for in the empagliflozin 10 and 25 mg groups, respectively; P < .0001) with both doses of empagliflozin than with placebo. There were also significantly larger decreases from baseline in fasting plasma glucose, 2-hour postprandial glucose and body weight with both empagliflozin doses than with placebo. Among patients in the empagliflozin 10 mg, 25 mg and placebo groups, 17.8%, 9.6% and 11.0% reported confirmed hypoglycaemic events, respectively (nominal P = .2422 and .7661 in the empagliflozin 10 and 25 mg groups, respectively), and no Clinical Events Committee-confirmed diabetic ketoacidosis events were reported. CONCLUSIONS: In Chinese patients with T2D, empagliflozin combined with insulin ± OADs improved glycaemic control and was well tolerated, without an increased risk of hypoglycaemia.

Long-term follow-up after discharge witnesses a slow decline of insulin autoantibodies in patients with insulin autoimmune syndrome complicated with Grave's disease: a report of two cases.

BACKGROUND: Insulin autoimmune syndrome (IAS) is a rare cause of hypoglycemia characterized by high levels of blood insulin autoantibodies. It has been documented that drugs containing sulfhydryl groups may result in IAS. In this study, we present two cases of IAS induced by methimazole, along with their corresponding treatments and a long-term follow-up after hospitalization. CASE PRESENTATION: We report two patients with Grave's disease (GD), carrying the HLA-DRB1 04:06 genotype, who experienced hypoglycemic episodes after taking methimazole. Inpatient treatments helped return their blood glucose levels to normal. Although no recurrences of hypoglycemia were present in the two cases studied, insulin autoantibodies remained positive for the previous follow-up sessions, which turned negative only three years after discharge. CONCLUSIONS: GD patients who carry the HLA-DRB1 04:06 genotype are prone to IAS if they take drugs containing sulfhydryl groups. It may take time for the elimination of insulin autoantibodies after the recovery from the hypoglycemic episode in IAS patients.

Pancreatic neuroendocrine tumor with ectopic adrenocorticotropic hormone syndrome: a case report and 5-year follow-up.

Pancreatic neuroendocrine tumors (P-NETs) secreting ectopic adrenocorticotropic hormones (ACTH) are rare and often delayed in diagnosis due to their atypical clinical characteristics. Here, we describe a case of P-NET in the pancreatic tail. The tumor had metastasized to the liver and secreted gastrin and ACTH. A 60-year-old female patient was diagnosed with gastrinoma in the pancreatic tail with liver metastases in 2015. After 3 months, the patient presented refractory hypokalemia and thyroid dysfunction. The final diagnosis was P-NET with ectopic ACTH syndrome (EAS). After cytoreductive surgery and the use of long-acting somatostatin analogs, plasma potassium levels and thyroid function were effectively corrected. Although Sandostatin LAR® Depot and proton pump inhibitors (PPIs) were used throughout the follow-up period, the tumor relapsed 4 years later. After aggressive treatment, including right hepatectomy, microwave coagulation of the left liver, and cholecystectomy, the tumor returned 4 months later. Finally, the patient underwent three hepatic artery embolizations and 12 courses of CAPTEM regimen chemotherapy. The markers of disease were almost maintained in the normal ranges until now. We have followed up on this case for more than 5 years. A timely and comprehensive examination of hormones and immunohistochemistry is essential. The prognosis of P-NET is poor. Regular long-term follow-up and the application of combined therapies are helpful to control the disease and improve the prognosis.

Efficacy and safety of PEGylated exenatide injection (PB-119) in treatment-naive type 2 diabetes mellitus patients: a Phase II randomised, double-blind, parallel, placebo-controlled study.

AIMS/HYPOTHESIS: Glucagon-like peptide 1 receptor agonists (GLP-1 RA) such as exenatide are used as monotherapy and add-on therapy for maintaining glycaemic control in patients with type 2 diabetes mellitus. The current study investigated the safety and efficacy of once-weekly PB-119, a PEGylated exenatide injection, in treatment-naive patients with type 2 diabetes. METHODS: In this Phase II, randomised, placebo-controlled, double-blind study, we randomly assigned treatment-naive Chinese patients with type 2 diabetes in a 1:1:1:1 ratio to receive subcutaneous placebo or one of three subcutaneous doses of PB-119 (75, 150, and 200 µg) for 12 weeks. The primary endpoint was the change in HbA1c from baseline to week 12, and other endpoints were fasting plasma glucose, 2 h postprandial glucose (PPG), and proportion of patients with HbA1c < 53 mmol/mol (<7.0%) and ≤48 mmol/mol (≤6.5%) at 2, 4, 8 and 12 weeks of treatment. Safety was assessed in all patients who received at least one dose of study drug. RESULTS: We randomly assigned 251 patients to one of the four treatment groups (n = 62 in placebo and 63 each in PB-119 75 µg, 150 µg and 200 µg groups). At the end of 12 weeks, mean differences in HbA1c in the treatment groups were -7.76 mmol/mol (95% CI -9.23, -4.63, p < 0.001) (-0.72%, 95% CI -1.01, -0.43), -12.89 mmol/mol (95% CI -16.05, -9.72, p < 0.001) (-1.18%, 95% CI -1.47, -0.89) and -11.14 mmol/mol (95% CI -14.19, -7.97, p <0 .001) (-1.02%, 95% CI -1.30, -0.73) in the 75 µg, 150 µg and 200 µg PB-119 groups, respectively, compared with that in the placebo group after adjusting for baseline HbA1c. Similar results were also observed for other efficacy endpoints across different time points. There was no incidence of treatment-emergent serious adverse event, severe hypoglycaemia or death. CONCLUSIONS/INTERPRETATION: All tested PB-119 doses had superior efficacy compared with placebo and were safe and well tolerated over 12 weeks in treatment-naive Chinese patients with type 2 diabetes. TRIAL REGISTRATION: ClinicalTrials.gov NCT03520972 FUNDING: The study was funded by National Major Scientific and Technological Special Project for Significant New Drugs Development and PegBio.

Henagliflozin as add-on therapy to metformin in patients with type 2 diabetes inadequately controlled with metformin: A multicentre, randomized, double-blind, placebo-controlled, phase 3 trial.

AIM: To evaluate the efficacy and safety of henagliflozin in patients with type 2 diabetes mellitus (T2DM) inadequately controlled with metformin. MATERIAL AND METHODS: This multicentre phase 3 trial included a 24-week randomized, double-blind, placebo-controlled period, followed by a 28-week extension period. Patients with a glycated haemoglobin (HbA1c) level of 7.0% (53 mmol/mol) to 10.5% (91 mmol/mol) were randomized and treated with once-daily placebo (n = 161), henagliflozin 5 mg (n = 162), or henagliflozin 10 mg (n = 160). After 24 weeks, patients on placebo were switched to 5 mg or 10 mg henagliflozin for the additional 28-week treatment, and patients on henagliflozin during 24-week treatment period maintained this initial therapy. The primary endpoint was change in HbA1c from baseline to Week 24. RESULTS: At Week 24, the least squares mean HbA1c changes versus placebo from baseline were - 0.76% (-8.3 mmol/mol) and - 0.80% (-8.7 mmol/mol) for henagliflozin 5 and 10 mg, respectively (all P < 0.0001). Compared with the placebo group, both doses of henagliflozin lowered fasting plasma glucose, 2-hour postprandial plasma glucose, body weight and blood pressure, and increased the proportions of patients achieving HbA1c <7.0% (53 mmol/mol) at Week 24. The trends in these improvements were sustained over an additional 28 weeks. Slightly higher proportions of ketosis and presence of urine ketone bodies were observed in patients treated with henagliflozin compared to placebo at Week 24. No diabetic ketoacidosis or episodes of severe hypoglycaemia were reported. CONCLUSIONS: Henagliflozin 5 mg or 10 mg as add-on therapy to metformin provided a new therapeutic option for the treatment of T2DM patients who have inadequate glycaemic control with metformin alone, and was generally well tolerated.

Changes of intestinal microflora of breast cancer in premenopausal women.

Breast cancer is one of the most common malignant tumors in women. More than half of breast cancer patients are not menopausal at the time of diagnosis. The occurrence and development of premenopausal breast cancer are affected by many factors. Intestinal flora, especially SCFA-producing bacteria, participates in the development of various tumors, and there is a lack of in-depth research in premenopausal breast cancer patients. We used 16S rRNA gene sequencing, targeted metabolomics, and cell culture methods to analyze the changes in the intestinal flora and metabolites of premenopausal breast cancer patients. In addition, we treated breast cancer cells with significantly altered propionate and butyrate in vitro to examine their effects on cell activity. This study followed STROBE guidelines. We found that compared with healthy premenopausal women, the composition and symbiosis of intestinal flora in patients with premenopausal breast cancer changed significantly. The abundance of short-chain fatty acid (SCFA)-producing bacteria was significantly reduced, and the key SCFA-producing enzymes were also significantly reduced. Pediococcus and Desulfovibrio could distinguish premenopausal breast cancer patients from normal premenopausal women. The related propionate and butyrate had a certain ability to inhibit breast cancer cell viability in vitro. As SCFA-producing bacteria, Pediococcus and Desulfovibrio showed potential reference value for the diagnosis of premenopausal breast cancer. The ability of propionate and butyrate to inhibit breast cancer cell lines in vitro suggests that the relevant SCFA receptor may be a new target for the treatment of premenopausal breast cancer.

miR-217 alleviates high-glucose-induced vascular smooth muscle cell dysfunction via regulating ROCK1.

MicroRNA-217 (miR-217) has been recently reported to be abnormally expressed during atherosclerosis. Nonetheless, it still remains unknown whether miR-217 can regulate inflammation, proliferation, migration, and apoptosis of vascular smooth muscle cells (VSMCs) in high-glucose condition. Sprague Dawley rats were used for establishing diabetic animal models. miR-217 mimics and miR-217 inhibitors were transfected into VSMCs. The miR-217 and ROCK1 expressions were measured by quantitative reverse transcription-polymerase chain reaction and Western blot. VSMCs' proliferation, migration, cell cycle, and apoptosis were validated using the Cell Counting Kit-8 assay, Transwell assay, and flow cytometry analysis, respectively. The binding sites between miR-217 and the 3'-untranslated region of ROCK1 were predicted via miRanda, PicTar, TargetScan, and microT databases, and the targeting relationship was confirmed by dual-luciferase reporter experiments. miR-217 was found to be upregulated in VSMCs treated by high glucose and aorta VSMCs of diabetic rats. Transfection of miR-217 mimics significantly induced VSMCs cycle arrest, inhibition of proliferation, reduction of migration, and enhancement of apoptosis. The bioinformatics analysis and dual-luciferase reporter experiments identified ROCK1 as a direct target of miR-217. miR-217 inhibits excessive proliferation and migration of VSMCs induced by high glucose by targeting ROCK1.

Metformin alleviates renal injury in diabetic rats by inducing Sirt1/FoxO1 autophagic signal axis.

Diabetic nephropathy (DN) is the major microvascular complication of diabetes mellitus and the most important cause of end-stage renal disease worldwide. Metformin is the preferred oral hypoglycaemic drug for type 2 diabetes mellitus (T2DM). Recent studies have shown that besides lowering blood glucose, metformin also has protective effects on renal function, but its mechanism is not clear. In this study, we established a diabetic rat model by high-fat feeding combined with intraperitoneal injection of streptozotocin. Their changes of renal function, oxidative stress, histopathology and structure, and autophagy were observed after 8 weeks of metformin treatment at different dose. Sirt1 inhibitor EX527 and metformin were used to observe whether the protective effect of metformin on DN kidney was achieved through the Sirt1/FoxO1 autophagic signalling pathway. The results showed that metformin could protect renal function by up-regulating autophagy level, alleviating oxidative stress level of renal tissue and pathological and structural changes of glomeruli, and inhibiting the expression of extracellular matrix. Sirt1 inhibitor could block the protective effect of metformin on kidney of diabetic rats, suggesting that metformin could alleviate kidney injury in diabetic rats by inducing Sirt1/FoxO1 autophagy signal axis. So metformin could alleviate renal injury in diabetic rats, which may be achieved by regulating Sirt1/FoxO1 autophagic signalling pathway and inducing renal autophagy.

Circulating apelin, chemerin and omentin levels in patients with gestational diabetes mellitus: a systematic review and meta-analysis.

BACKGROUND: The available data on the significance of circulating apelin, chemerin and omentin in women with gestational diabetes mellitus (GDM) are inconsistent. This analysis includes a systematic review of the evidence associating the serum concentrations of these adipokines with GDM. METHODS: Publications through December 2019 were retrieved from PubMed, Embase, the Cochrane Library, and Web of Science. Subgroup analysis and meta-regression were conducted to evaluate sources of heterogeneity. RESULTS: Analysis of 20 studies, including 1493 GDM patients and 1488 normal pregnant women did not find significant differences in circulating apelin and chemerin levels (apelin standardized mean difference [SMD] = 0.43, 95% confidence interval (CI): - 0.40 to 1.26, P = 0.31; chemerin SMD = 0.77, 95% CI - 0.07 to 1.61, P = 0.07). Circulating omentin was significantly lower in women with GDM than in healthy controls (SMD = - 0.72, 95% CI - 1.26 to - 0.19, P = 0.007). Publication bias was not found; sensitivity analysis confirmed the robustness of the pooled results. CONCLUSIONS: Circulating omentin was decreased in GDM patients, but apelin and chemerin levels were not changed. The results suggest that omentin has potential as a novel biomarker for the prediction and early diagnosis of GDM.

Circulating FABP4, nesfatin-1, and osteocalcin concentrations in women with gestational diabetes mellitus: a meta-analysis.

OBJECTIVE: Recent studies have investigated the circulating adipocyte fatty acid binding protein (FABP4), nesfatin-1, and osteocalcin (OC) concentrations in women diagnosed with gestational diabetes mellitus (GDM), but the findings prove to be conflicting. The objective of this research was to systematically assess the relationship of circulating levels of above adipokines with GDM. METHODS: Pubmed, Embase, Web of Science, Cochrane library, OVID, and Scopus were performed to locate articles published up to January 31, 2020. Pooled standard mean differences (SMDs) with 95% confidence intervals (CIs), and 95% predictive intervals (PIs) were calculated by random-effects models to compare levels of adipokines between GDM cases and control groups. Cumulative and single-arm meta-analyses were also performed. RESULTS: Thirty-one studies comprising 4590 participants were included. No significant differences were found between GDM women and healthy controls in circulating nesfatin-1 levels (4.56 vs. 5.02 ng/mL; SMD = - 0.11, 95% CI -0.61-0.38, 95% PI -1.63-1.41). Nevertheless, circulating FABP4 and OC levels observed in GDM women outnumbered normal controls (FABP4, 23.68 vs. 16.04 ng/mL; SMD = 2.99, 95% CI 2.28-3.69, 95% PI 0.28-5.71; OC, 52.34 vs. 51.04 ng/mL; SMD = 0.68, 95% CI 0.31-1.05, 95% PI -0.48-1.84). The cumulative meta-analysis showed that the SMDs of circulating FABP4 and OC levels had stabilized between the two groups. CONCLUSIONS: Elevated circulating FABP4 and OC levels were observed in GDM women, but nesfatin-1 levels did not change, the PI of OC crossed the no-effect threshold. The results suggested that FABP4 is more suitable as a biomarker of GDM compared to OC in a future study, which is useful in identifying pregnant women who are likely to develop GDM and providing prompt management strategies.

CREB-upregulated lncRNA MEG3 promotes hepatic gluconeogenesis by regulating miR-302a-3p-CRTC2 axis.

Hepatic gluconeogenesis is the major contributor to hyperglycemia in diabetes. Long noncoding RNA (lncRNA) maternally expressed gene 3 (MEG3) has been shown to promote hepatic insulin resistance; however, the underlying mechanism involving hepatic gluconeogenesis remains unclear. This study aims to investigate the potential role of MEG3 in hepatic gluconeogenesis. Mouse primary hepatocytes were used in this study. Cell transfection was performed for the overexpression or knockdown of specific genes. Expressions of MEG3, miR-302a-3p, CREB-regulated transcriptional coactivator 2 (CRTC2), protein kinase A (PKA), cAMP-response element binding protein (CREB), PPARγ coactivator-1α (PGC-1α), phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6-phosphatase (G6Pc) were determined by quantitative real-time polymerase chain reaction (qRT-qPCR) and Western blot analysis, respectively. The association among MEG3, miR-302a-3p, and CRTC2 was disclosed by dual-luciferase reporter assay. MEG3 was highly expressed in high glucagon-treated mouse primary hepatocytes. CREB-induced MEG3 upregulation increased gluconeogenic gene expression in high glucagon-treated primary hepatocytes, while MEG3 interference led to an opposite effect. MEG3 served as a competing endogenous RNA (ceRNA) to upregulate CRTC2 by targeting miR-302a-3p in primary hepatocytes, thereby increasing PGC-1α-PEPCK/G6Pc. CREB-upregulated MEG3-enhanced hepatic gluconeogenesis via mediating miR-302a-3p-CRTC2 axis, revealing that MEG3 might be a potential target and therapeutic strategy for diabetes.

Metformin inhibits high glucose-induced mesangial cell proliferation, inflammation and ECM expression through the SIRT1-FOXO1-autophagy axis.

The aim of this study was to investigate the role of metformin in high glucose-induced mesangial cell proliferation, inflammation and extracellular matrix (ECM) accumulation and to elucidate the underlying mechanism of metformin function. An MTT assay was used to examine rat mesangial cell (RMC) proliferation. The levels of TNF-α, IL-6 and TGF-ß in RMCs were determined by ELISA. The protein expression of fibronectin, collagen IV and autophagy-related proteins (Beclin-1, LC3-I and LC3-II) in RMCs was detected using western blot. Fluorescence microscopy analysis was carried out to evaluate RMC autophagy. Our results showed that high glucose-induced RMC proliferation, inflammation and ECM expression, but these effects were markedly reduced by metformin. We confirmed that metformin suppressed high glucose-induced RMC proliferation, inflammation and ECM expression via induction of autophagy. Mechanistic investigation demonstrated an axis of SIRT1-FOXO1 in RMC autophagy. Our data indicated that metformin inhibits high glucose-induced mesangial cell proliferation, inflammation and ECM expression through a SIRT1-FOXO1-autophagy axis.

The association between thyroid dysfunction (TD) and diabetic kidney disease (DKD) in type 2 diabetes mellitus (T2DM).

BACKGROUND: Epidemiological studies have identified an association between thyroid dysfunction (TD) and various kidney diseases. In this study, the prevalence of TD in type 2 diabetic mellitus (T2DM) patients with diabetic kidney disease (DKD) was evaluated to analyse the potential association between TD and DKD in T2DM patients. METHODOLOGY: A total of 2108 T2DM patients from Anhui Provincial Hospital were recruited in this study. Demographic and clinical characteristics were collected from 834 T2DM patients with DKD and 1274 T2DM patients without DKD (non-DKD). All patients were stratified into a number of groups based on UACR (urine albumin-to-creatinine ratio) or eGFR (estimated glomerular filtration rate): (a) A1: normoalbuminuria (<30), A2: microalbuminuria (30-300) and A3: macroalbuminuria (>300); (b) F1: normal filtration (60-139), F2: hyper filtration (≥140) and F3: low filtration (<60). RESULTS: Significant differences were observed between the non-DKD and DKD groups (P < .05) in age, sex ratio, duration, systolic blood pressure, total cholesterol, low density lipoprotein cholesterol, serum creatinine, blood urea nitrogen, free triiodothyronine (FT3), free thyroxine (FT4) and sensitive thyrotropin hormone (sTSH). The macroalbuminuira and low filtration groups had the lowest levels of FT3 and FT4 and the highest level of sTSH, compared with all other groups (P < .0167). The prevalence of subclinical hypothyroidism in the DKD group was significantly higher than that in the non-DKD group (χ2  = 13.92, P < .01). Logistic regression analysis showed that hypothyroidism was associated with increased UACR or reduced eGFR in T2DM patients. Compared with controls, T2DM patients with hypothyroidism exhibited a higher UACR and urinary excretion of transferrin, as well as a lower excretion of urinary Tamm-Horsfall protein (THP) (P < .0167). CONCLUSION: Subclinical hypothyroidism is more prevalent in T2DM patients with DKD than in T2DM patients without DKD. Hypothyroidism is associated with albuminuria and decreased eGFR in T2DM patients.

Rapamycin improves insulin resistance and hepatic steatosis in type 2 diabetes rats through activation of autophagy.

Insulin resistance (IR) is a hallmark of type 2 diabetes mellitus (T2DM). This study aimed to explore the effects of rapamycin, a specific inhibitor of kinase mammalian target of rapamycin (mTOR), on IR in T2DM rats, and to validate whether the underlying mechanism was associated with autophagy. In this study, the model of T2DM rats was established by feeding the animals with a high-fat diet (HFD) and intraperitoneal injection of streptozotocin (STZ). Diabetic rats were randomly divided into model of T2DM control group (DM-C, n = 15), metformin group (DM-M, n = 15), rapamycin group (DM-Rapa, n = 15), 3-methyladenine (3-MA) group (DM-3-MA, n = 15), and rapamycin + 3-MA group (DM-Rapa-3-MA, n = 15). Rats in different treatment groups were given by corresponding therapy from gastric tube. Meanwhile, normal control group was established (n = 10). As expected, HFD- and STZ- induced T2DM rats exhibited significantly impaired glucose tolerance, reduced insulin sensitivity, dysglycemia and dyslipidemia, aggravated hepatic steatosis, enhanced hepatic inflammation, elevated p-mTOR, and suppressed hepatic autophagy. Importantly, rapamycin and metformin significantly ameliorated IR, relieved disorders of glucose and lipid metabolism, reduced inflammatory level, inhibited mTOR, and promoted autophagy. Importantly, the autophagy inhibitor 3-MA significantly reversed the effects exerted by rapamycin. Collectively, our study suggests that rapamycin improved IR and hepatic steatosis in T2DM rats via activation of autophagy.

A study of urinary Tamm-Horsfall protein excretion in adult type 2 diabetes mellitus
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AIM: To study the excretion of urinary Tamm-Horsfall protein (THP) with a wide range of urinary albumin creatinine ratio (UACR) and estimated glomerular filtration rate (eGFR) in a population with type 2 diabetes mellitus (T2DM). MATERIALS AND METHODS: A retrospective investigation was performed in 3,599 adult inpatients with T2DM in Anhui Provincial Hospital of China. Demographic information, urinary THP data, and other lab reports were obtained from the Electronic Patient Record (EPR). RESULTS: (1) In T2DM, average HbA1c was 8.6%, and 55.2% of the patients had an HbA1c > 9%. The percentages with normal excretion, decreased excretion, and increased excretion of urinary THP were 49.8%, 41.7%, and 8.5%, respectively. Serum creatinine (Scr), blood urea nitrogen (BUN), and UACR levels were highest, and eGFR was lowest, in the decreased urinary THP group. (2) The proportion of decreased urinary THP excretion rose, and the level of urinary THP declined when UACR increased or eGFR decreased. (3) Urinary THP showed a negative correlation to Scr, BUN, triglyceride (TG), and UACR (rs = -0.173, -0.228, -0.060 and -0.237, respectively), but a positive correlation with eGFR and high-density lipoprotein cholesterol (HDL-C) (rs =0.128 and 0.040, respectively). (4) Logistical regression analysis showed that Scr, BUN, TG, UACR, and eGFR were independent risk factors for decreased urinary THP (OR = 3.477, 1.461, 1.160, 2.124, and 2.087, respectively). CONCLUSION: Increased albuminuria and decreased GFR are predictors of decreased urinary THP excretion. Distal convoluted tubule lesion may precede glomerular damage in a portion of T2DM patients.
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MiR-181c restrains nitration stress of endothelial cells in diabetic db/db mice through inhibiting the expression of FoxO1.

Endothelial dysfunction played an important role in the progression of diabetes mellitus (DM). miR-181c has been implicated in many diseases, including DM. However, the molecular mechanisms of miR-181c regulate this process remained poorly understood. Healthy ICR mice were divided into control group (n = 10) and db/db DM group (n = 10). The expression of miR-181c and FoxO1 were both investigated in diabetic db/db mice or high glucose-induced endothelial cells (MAECs and END-D). Here we found that down-regulation of miR-181c and the activation of FoxO1/iNOS were observed in mice and endothelial cells. Furthermore, we verified that miR-181c directly targeted and inhibited FoxO1 gene expression by targeting its 3'-UTR through luciferase reporter assay. Knockdown of FoxO1 reversed the up-regulation of iNOS, nitrotyrosine and the down-regulation of p-eNOSSer1177/eNOS in high glucose (30 mM)-induced MAECs cells. In addition, over-expression of miR-181c could reverse the enhanced nitration stress induced by high glucose, while this effect could be attenuated by pcDNA-FoxO1 in MAECs. These results shown that miR-181c attenuated nitration stress through regulating FoxO1 expression and affecting endothelial cell function, which offering a new target for the development of preventive or therapeutic agents against DM.

Comparison of Antidiabetic Medications during the Treatment of Atherosclerosis in T2DM Patients.

Type 2 diabetes is often associated with arterial atherosclerosis in large blood vessels. We set out to elucidate whether commonly used antidiabetic drugs metformin, sitagliptin, and pioglitazone will reduce atherosclerosis in T2DM patients. We enrolled 176 individuals with type 2 diabetes, which were divided into four treatment groups according to different oral drugs: metformin alone, sitagliptin alone, pioglitazone alone, or combination of metformin and sitagliptin. We assessed changes in glycometabolism, lipid metabolism, cytokine released, and carotid artery intima-media thickness as the readout for improvement in atherosclerosis. HbA1c levels were significantly decreased in all treatment groups (p < 0.05), and FBG levels were also decreased in metformin and combined groups (p < 0.05). In addition, we found IL-6 levels significantly decreased in all treatment groups (p < 0.05). Treatment with pioglitazone showed a significant increase in BMI, HDL, and ADPN levels (p < 0.05). We also observed a significant decrease in NHDL levels in the combined treatment group (p < 0.05). Our data revealed that in addition to hypoglycemic properties of metformin, sitagliptin, and pioglitazone, these drugs also have the potential to promote an anti-inflammatory response. Therefore, combination therapy may be more beneficial for reducing atherosclerosis in patients with type 2 diabetes. The clinical trial is registered with ChiCTR-ORC-17010835.

[Protective effects of metformin on glomerular podocytes of type 2 diabetes model rats].

OBJECTIVE: To observe the effects of metformin (MET) on podocalyxin (PCX) expression in renal tissue from type 2 diabetes mellitus (T2DM) model rats and investigate its protective effects against glomerular podocyte injury. METHOD: The rat model of T2DM was established by feeding with high-fat diet and intraperitoneal injection of low dose of streptozotocin (STZ). All the rats were divided into four groups: diabetic group (n=9), metformin group (300 mg·kg(-1)·d(-1), n=8), glibenclamide group (5 mg·kg(-1)·d(-1), n=8 ) and normal group (n=8). After 8 weeks, urinary PCX and creatinine, blood glucose (BG) and glycated hemoglobin (HbA1c) were detected in all the rats. Immunohistochemistry was used to observe the protein expression of PCX in renal tissue. Real-time polymerase chain reaction (PCR) was performed to detect mRNA expression of PCX. Pathological changes of renal tissue were observed by electron microscope. RESULTS: Metformin and glyburide treatment decreased the levels of BG and HbA1c [(9.6±1.1) and (9.9±1.1) vs (15.6±1.6) mmol/L, (7.0±0.3)% and (8.0±1.0)% vs (12.4±0.6)%, all P<0.05], compared with diabetes group, while there was no statistically significant difference between the two intervention groups (P>0.05). The level of urinary PCX/urinary creatinine (UPCR) in diabetic rats were higher than that of normal group [(697±136) vs (94±25 ) ng/g, P<0.05), meanwhile the levels of protein and mRNA expression of PCX in kidney tissue reduced remarkably [(0.75±0.11) vs (3.18±0.14), (0.08±0.09) vs (1.00±0.02), both P<0.05]. Above-mentioned indicators could be ameliorated by metformin and glyburide treatment compared to normal group (P<0.05), and there were statistically significant difference between the two intervention groups [(404±83) vs (516±38) ng/g, (1.54±0.06) vs(1.06±0.10), (0.23±0.01) vs (0.16±0.04), all P<0.05)]. Further observation found that basement membrane thickness of kidney [(267±22) vs (106±10)nm )] and fusion rate of foot process (0.80±0.07 vs 0) increased in the rats of diabetic group, and metformin or glyburide treatment can significantly reduced the above changes (P<0.05), additionally, the changes were remarkable different between metformin and glyburide group [(151±17) vs (204±22 ) nm, (0.49±0.04) vs (0.57±0.03), both P<0.05)]. CONCLUSION: Metformin has protective effect on glomerular podocytes by regulating the expression of PCX in renal tissue, independent of its hypoglycemic effect.