12-hydroxy-5-8-10-14-eicosatetraenoic-acid and Diabetes-Mellitus--Type-2

To investigate the relationship between serum 12-Hydroxyeicosatetraenoic acid (12-HETE) and diabetic retinopathy (DR) in children with type 1 diabetes mellitus (T1DM) and adults with type 2 diabetes mellitus (T2DM).. Children from the Shanghai Children and Adolescent Diabetes Eye (SCADE) study and adults from the Shanghai Cohort Study of Diabetic Eye Disease (SCODE) were examined in 2021. Serum 12-HETE levels were detected and compared. Multivariate logistic regression was used to analyze the relationship between 12-HETE and the rate of DR in diabetic patients.. The child study included 4 patients with new-onset DR and 24 patients with T1DM without DR. In children with T1DM, the 12-HETE level was significantly higher in those with DR (P = 0.003). The adult study had two sets, for testing and verification. The test set included 28 patients with new-onset DR and 24 T2DM patients with a course of ≥ 20 years who had never developed DR. The verification set included 41 patients with DR, 50 patients without DR and 50 healthy controls. In the adult test set, the 12-HETE level was significantly higher in patients with DR than in those with T2DM without DR (P = 0.003). In the verification set, the 12-HETE level of patients with DR was significantly higher than that of patients without DR (P < 0.0001) and the healthy controls (P < 0.0001). Multivariate logistic regression indicated that 12-HETE was independently associated with DR in both children (odds ratio [OR] 1.06, 95% confidence interval [CI] 1.00-1.13, P = 0.041) and adults (test set [OR 9.26, 95% CI 1.77-48.59, P = 0.008], verification set [OR 10.49, 95% CI 3.23-34.05, P < 0.001]).. Higher serum 12-HETE levels are positively correlated with an increased risk of DR in children with T1DM and adults with T2DM.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Adolescent; Adult; Child; China; Cohort Studies; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Humans; Risk Factors

Heart failure with preserved ejection fraction (HFpEF) is associated with endothelial dysfunction and is frequent in people with type 2 diabetes mellitus. In diabetic patients, increased levels of the eicosanoid 12-hydroxyeicosatetraenoic acid (12-HETE) are linked to vascular dysfunction. Here, we aimed to identify the importance of 12-HETE in type 2 diabetic patients exhibiting diastolic dysfunction, and mice exhibiting HFpEF and whether targeting 12-HETE is a means to ameliorate HFpEF progression by improving vascular function in diabetes.. Endothelium-dependent vasodilation and mitochondrial functional capacity both improved in response to either application of ML351 or the V1-cal peptide. Correlating to improved vascular function, mice treated with either pharmacological agent exhibited improved diastolic filling and left ventricular relaxation that correlated with increased myocardial capillary density.. Our results suggest that 12-HETE may serve as a biomarker indicating endothelial dysfunction and the resulting cardiovascular consequences such as HFpEF in type 2 diabetic patients. Antagonizing 12-HETE is a potent means to causally control HFpEF development and progression in type 2 diabetes by preserving vascular function.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Aged; Animals; Blood Vessels; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diastole; Endothelial Cells; Female; Heart Failure; Heart Ventricles; Humans; Male; Mice; Mitochondria; Stroke Volume; TRPV Cation Channels; Vasodilation

The enteric nervous system (ENS) plays a key role in controlling the gut-brain axis under normal and pathological conditions, such as type 2 diabetes. The discovery of intestinal actors, such as enterosynes, able to modulate the ENS-induced duodenal contraction is considered an innovative approach. Among all the intestinal factors, the understanding of the role of gut microbes in controlling glycaemia is still developed. We studied whether the modulation of gut microbiota by prebiotics could permit the identification of novel enterosynes.. We measured the effects of prebiotics on the production of bioactive lipids in the intestine and tested the identified lipid on ENS-induced contraction and glucose metabolism. Then, we studied the signalling pathways involved and compared the results obtained in mice to human.. We found that modulating the gut microbiota with prebiotics modifies the actions of enteric neurons, thereby controlling duodenal contraction and subsequently attenuating hyperglycaemia in diabetic mice. We discovered that the signalling pathway involved in these effects depends on the synthesis of a bioactive lipid 12-hydroxyeicosatetraenoic acid (12-HETE) and the presence of mu-opioid receptors (MOR) on enteric neurons. Using pharmacological approaches, we demonstrated the key role of the MOR receptors and proliferator-activated receptor γ for the effects of 12-HETE. These findings are supported by human data showing a decreased expression of the proenkephalin and MOR messanger RNAs in the duodenum of patients with diabetic.. Using a prebiotic approach, we identified enkephalin and 12-HETE as new enterosynes with potential real beneficial and safety impact in diabetic human.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Adult; Aged; Animals; Blood Glucose; Brain-Gut Axis; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Duodenum; Enkephalins; Enteric Nervous System; Gastrointestinal Microbiome; Glucose Tolerance Test; Humans; Isotonic Contraction; Male; Mice; Middle Aged; Muscle, Smooth; Neurons; Nitric Oxide Synthase Type I; Oligosaccharides; PPAR gamma; Prebiotics; Protein Precursors; Receptors, Opioid, mu; RNA, Messenger; Signal Transduction

The 12-lipoxygenase (12-LO) and angiotensin II (Ang II) interaction plays an important role in diabetic nephropathy (DN). Proteinuria in DN is associated with decreased slit diaphragm proteins including nephrin and P-cadherin. Therefore, we investigated whether Ang II type 1 receptor (AT1) blocker (ARB) regulates 12-LO activity and slit diaphragm protein expression in diabetic rat glomeruli.. Glomeruli were isolated with the sieving method, and classified into small glomeruli (SG; 75-μm sieve) and large glomeruli (LG; 125-μm sieve).. 12(S)-HETE, a lipid product of 12-LO, was increased by Ang II in the glomeruli. Infusion of 12(S)-HETE and Ang II significantly decreased nephrin expression in LG, but increased it in SG compared to control. Glomerular P-cadherin expression was reduced after Ang II and 12(S)-HETE treatment without differences between LG and SG. ARB did not influence glycemic levels but completely abolished the increases in 12(S)-HETE, AT1 expression, and proteinuria in diabetic rats. Nephrin expression was significantly reduced in LG but increased in SG in diabetic rats compared to control. P-cadherin expression decreased in both diabetic LG and SG. The abnormalities of nephrin and P-cadherin were partially but significantly reversed by ARB.. ARB potentially ameliorates DN via the up-regulation of glomerular nephrin and P-cadherin expression through the inhibition of 12-LO activation in the glomeruli of rats with DN.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Angiotensin II; Angiotensin II Type 1 Receptor Blockers; Animals; Arachidonate 12-Lipoxygenase; Cadherins; Cells, Cultured; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Diet, High-Fat; Kidney Glomerulus; Lipoxygenase Inhibitors; Losartan; Male; Membrane Proteins; Mice; Podocytes; Rats, Sprague-Dawley; Receptor, Angiotensin, Type 1; Renin-Angiotensin System; Streptozocin

(1) BACKGROUND: 12-lipoxygenase (12-LO) is involved in the development of diabetic nephropathy (DN). In the present study, we investigated whether 12-LO inhibition may ameliorate type-2 DN (T2DN) by interfering with insulin resistance (IR); (2) METHODS: Rat glomerular mesangial cells, glomeruli and skeletal muscles were isolated and used in this study. Kidney histological changes were confirmed by periodic-acid Schiff staining; mRNA expression was detected by competitive reverse transcription polymerase chain reaction; and the protein level was determined by Western blot and the enzyme-linked immunosorbent assay, respectively; (3) RESULTS: The inhibition of 12-LO attenuated microalbuminuria (MAU) increases in type-2 diabetic rats, but not in type-1 diabetic rats. Infusion of 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) significantly increased the expression of angiotensin II (Ang II) and Ang II type 1 receptor (AT1R), but decreased the expression of AT1R-associated protein (ATRAP) in rat glomeruli, compared to the control. An in vitro study revealed that both 12(S)-HETE and insulin upregulated AT1R expression in rat mesangial cells. In the presence of p38 mitogen-activated protein kinase (MAPK) inhibitor, SB202190, the 12(S)-HETE-induced ATRAP reduction was significantly abolished. Interestingly, 12-LO inhibition did not influence AT1R expression in type-1 diabetic rats, but significantly abolished the increased AT1R and Ang II expression in glomeruli of type-2 diabetic rats. Furthermore, the inhibition of 12-LO significantly corrected impaired insulin sensitivity and fast serum insulin level, as well as the p-AMP-activated protein kinase (AMPK) reduction in skeletal muscle of type-2 diabetic rats; (4) CONCLUSION: The inhibition of 12-LO potentially ameliorated MAU by preventing IR through the downregulation of glomerular AT1R expression in T2DN.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Albuminuria; Animals; Arachidonate 12-Lipoxygenase; Cells, Cultured; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Down-Regulation; Insulin Resistance; Kidney Glomerulus; Lipoxygenase Inhibitors; Male; Muscle, Skeletal; Protein Kinase Inhibitors; Rats; Rats, Sprague-Dawley; Receptor, Angiotensin, Type 1

Arachidonic acid-metabolizing enzyme, 12-lipoxygenase (12-LO), is involved in the glomerular hypertrophy of diabetic nephropathy (DN), in which cyclin-dependent kinase inhibitors (CKIs) play important roles. However, it is unclear whether 12-LO regulates the expression of the CKI p16(ink4a) in DN.. Primary glomerular mesangial cells (MCs) and glomeruli isolated from rats were used in this study. The rats were fed a high-fat diet and given low-dose streptozotocin to induce type 2 diabetes. The 12-LO product, 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE), was infused through an osmotic minipump. Enzyme-linked immunosorbent assay, Western blot, and morphometric analyses were performed.. High glucose (HG) increased the p16(ink4a) protein expression in MCs, but this increase was prevented by the 12-LO inhibitor, cinnamyl-3,​4-dihydroxy-α-cynanocinnamate (CDC). The levels of p-p38MAPK and p16(ink4a) in MCs were significantly elevated after the 12(S)-HETE treatment, whereas the p38MAPK inhibitor SB203580 prevented these increases. Compared with levels in control MCs, marked increases in p38MAPK activation and p16(ink4a) expression were observed in MCs plated on collagen IV, while the CDC treatment prevented these changes. Subcutaneous injection of CDC did not affect glucose levels, but completely attenuated the diabetes-related increases in the 12(S)-HETE content, p16(ink4a) expression, p-p38MAPK levels, glomerular volume, and the kidney/body weight ratio. Compared with levels in controls, p16(ink4a) and p-p38MAPK in the glomeruli derived from 12(S)-HETE-treated rats were significantly higher.. 12-LO-p38MAPK mediates the upregulation of p16(ink4a) in HG-stimulated MCs and type 2-diabetic glomeruli, and new therapies aimed at 12-LO inhibition may prove beneficial in ameliorating diabetes-induced glomerular hypertrophy.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Animals; Arachidonate 12-Lipoxygenase; Cells, Cultured; Collagen Type IV; Cyclin-Dependent Kinase Inhibitor p16; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Glomerular Mesangium; Glucose; p38 Mitogen-Activated Protein Kinases; Rats; Rats, Sprague-Dawley

12(S)-Hydroxyeicosatetraenoic acid (12(S)-HETE) is a metabolite of arachidonic acid. 12(S)-HETE is involved in the pathogenesis of atherosclerosis and diabetes. However, the correlation between 12(S)-HETE and coronary artery disease (CAD) in the diabetic patient is unclear.. The study investigated the relationship between 12(S)-HETE and CAD in Type 2 diabetes (T2D).. Plasma 12(S)- HETE levels were detected by enzyme-linked immunosorbent assay in 103 healthy controls (control), 109 diabetic patients without CAD (diabetic), and 152 diabetic patients with CAD (diabetic-CAD).. 12(S)-HETE levels were higher in both diabetic and diabetic-CAD groups compared to control and in the diabetic-CAD group compared to the diabetic group. In the multiple linear stepwise regression analysis, 12(S)-HETE levels correlated independently with CAD, systolic blood pressure, and glycated hemoglobin.. These results indicate that 12(S)-HETE levels are increased in diabetic patients with CAD, suggesting a role for atherosclerosis in T2D.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Adult; Aged; Blood Pressure; Case-Control Studies; Coronary Artery Disease; Diabetes Mellitus, Type 2; Diabetic Angiopathies; Female; Humans; Male; Middle Aged; Regression Analysis; Risk Factors

Angiotensin II type 1 receptor (AT1) plays an important role in the development of diabetic nephropathy (DN). However, the roles played by 12-lipoxygenase (12-LO) in the AT1 expression in glomerular cells exposed to high glucose (HG) and diabetic glomeruli remain unclear. Our objective in the present study was to investigate the role of 12-LO in the AT1 expression in glomerular cells and glomeruli under diabetic conditions.. Mesangial cells (MCs), podocytes and glomeruli isolated from rats were used in this study. The rats fed a high fat diet received low-dose streptozotocin to make type 2 diabetes. The 12-LO product 12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] was infused to rats by osmotic mini-pump. Morphometric measurement for glomerular volume, competitive reverse transcription polymerase chain reaction for mRNA expression, western blot and immunohistochemistry for protein expression were performed, respectively.. Both the 12(S)-HETE and HG increased AT1 protein expression in MCs and podocytes. Furthermore, the levels of the AT1 were significantly higher in glomeruli derived from 12(S)-HETE-treated rats compared with control rats. In addition, HG-induced AT1 expression was significantly reduced by the 12-LO inhibitor cinnamyl-3,4-dihydroxy-alpha-cynanocinnamate (CDC). Compared with the non-diabetic controls, DN rats showed significant glomerular hypertrophy and albuminuria. This was associated with significant increases in AT1 protein expression. These abnormalities were prevented by treatment of the CDC.. These results indicate that AT1 expression is enhanced, at least in part, by 12-LO in the type 2 diabetic glomeruli, and 12-LO inhibition can ameliorate DN progression through downregulation of AT1 expression.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Animals; Arachidonate 12-Lipoxygenase; Base Sequence; Cells, Cultured; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; DNA Primers; Gene Expression; Glucose; Hypertrophy; Kidney Glomerulus; Male; Mesangial Cells; Mice; p38 Mitogen-Activated Protein Kinases; Podocytes; Rats; Rats, Sprague-Dawley; Receptor, Angiotensin, Type 1

Arachidonic acid (AA) is a precursor of important vasoactive metabolites, but the role of AA-mediated vasodilation in Type 2 diabetes is not known. Using Zucker diabetic fatty (ZDF) rats, we examined the effects of AA in small mesenteric arteries preconstricted with endothelin. In ZDF rat mesenteric arteries, 1 microM AA produced only one-third the amount of dilation as in vessels from lean control animals. In lean control rats, the effect of AA was significantly and predominantly inhibited by the lipoxygenase inhibitors baicalein and cinnamyl-3,4-dihydroxy-cyanocinnamate (CDC). However, baicalein and CDC had no effect on AA-mediated dilation in ZDF rat mesenteric arteries. The major [3H]AA metabolite produced by isolated mesenteric arteries in both lean and ZDF rats was 12-hydroxyeicosatetraenoic acid (12-HETE), but the amount of [3H]12-HETE produced by ZDF rat vessels was only 36% of that of control vessels. In addition, 12-HETE produced similar amounts of dilation in lean and ZDF rat mesenteric arteries. Immunoblot analysis showed an 81% reduction in 12-lipoxygenase protein in ZDF rat mesenteric arteries. Immunofluorescence labeling showed strong nitrotyrosine signals in ZDF rat mesenteric arteries that colocalized with 12-lipoxygenase in endothelium, and 12-lipoxygenase coprecipitation with anti-nitrotyrosine antibodies was enhanced in ZDF rat vessels. We conclude that AA-mediated relaxation in ZDF rat small mesenteric arteries is impaired due to reduced 12-lipoxygenase protein and activity. Increased oxidative stress and nitration of 12-lipoxygenase may underlie the impairment of AA-mediated relaxation in small mesenteric arteries of diabetic rats.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Animals; Arachidonate 12-Lipoxygenase; Arachidonic Acid; Diabetes Mellitus, Type 2; Dose-Response Relationship, Drug; Eicosanoids; Male; Mesenteric Arteries; Nitrogen; Obesity; Potassium Channels; Rats; Rats, Zucker; RNA, Messenger; Tyrosine; Vasodilation

Earlier studies in diabetic animal models or ex vivo from diabetics suggest a deficiency in prostacyclin (PGI2) production and an increase in an alternate arachidonic acid metabolite, 12-hydroxyeicosatetraenoic acid (12-HETE), which stimulates angiogenesis, mitogenesis, and inhibits renin secretion. We studied the urinary excretion rate of 6-keto-PGF1 alpha (a stable metabolite of PGI2) and 12-HETE in controls and 42 noninsulin-dependent diabetes mellitus (NIDDM) patients with normal renal function and those with micro- or macroalbuminuria/hyporeninemic hypoaldosteronism (HH). The 2 eicosanoids were measured in urine using previously described high pressure liquid chromatography and RIA methods. Normal subjects and patients with NIDDM and microalbuminuria were infused with low dose calcium infusions that stimulate prostacyclin production in normal subjects. The PGI2 excretion rate of NIDDM patients with normal renal function was not different from that of controls (143 +/- 17 vs. 118 +/- 34 ng/g creatinine), but was reduced in those with microalbuminuria (75 +/- 10) and in macroalbuminuria patients (48 +/- 7; P < 0.01). In contrast, 12-HETE was increased in diabetics with normal renal function as well as in those with micro- or macroalbuminuria patients (69 +/- 18 vs. 250 +/- 62 vs. 226 +/- 60 and 404 +/- 131 ng/g creatinine; P < 0.01). Calcium did not stimulate PGI2, but increased 12-HETE in diabetics with microalbuminuria in contrast to levels in normal subjects. HH patients excreted less PGI2 (as previously reported), but had increased 12-HETE. HETE/PGI2 ratios further demonstrated these changes in the various groups. In a nondiabetic hypertensive microalbuminuria group, 12-HETE excretion was normal (73 +/- 28 ng/g creatinine). We conclude that the lipoxygenase product 12-HETE is increased early in the diabetic process, whereas PGI2 production is progressively impaired in NIDDM. These changes may play a role in the vascular disease of diabetes and partially explain the HH syndrome.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Adult; Aged; Albuminuria; Arachidonate 12-Lipoxygenase; Calcium; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Epoprostenol; Female; Humans; Hydroxyeicosatetraenoic Acids; Hypoaldosteronism; Male; Middle Aged; Renin