Irisin levels are associated with urotensin II levels in diabetic patients

Aims/Introduction

Irisin is a newly identified myokine that can promote energy expenditure. Previous studies showed that circulating urotensin II (UII) levels were increased in diabetes, and UII could inhibit the glucose transport in skeletal muscle in diabetic mice and aggravated insulin resistance. We presumed that irisin levels are associated with UII in diabetic patients.

Materials and Methods

A total of 71 patients with type 2 diabetes and 40 healthy subjects were recruited. Blood and urinary irisin concentrations were measured by using enzyme-linked immunosorbent assay, and UII concentrations were measured by bioelectrical impedance analysis. Every participant''s body composition was analyzed by bioelectrical impedance.

Results

The serum irisin levels were significantly lower in diabetic patients than that of controls, whereas serum UII levels were significantly higher in diabetic patients than that in that of controls. Serum irisin levels were negatively associated with circulating UII, hemoglobin A1c and the natural logarithm transformation of urinary albumin excretion, whereas serum irisin was positively associated with estimated glomerular filtration rate, and low-density lipoprotein cholesterol and urinary irisin were positively associated with urinary UII. Furthermore, circulating irisin is positively associated with muscle mass, whereas circulating UII is negatively associated with muscle mass in diabetic patients. Hemoglobin A1c and circulating UII are independent determinants of circulating irisin by multiple regression analysis.

Conclusions

The present results provide the clinical evidence of an association between irisin and UII in diabetic patients. Hemoglobin A1c and circulating UII are independent determinants of circulating irisin. Our results hint that UII and high glucose might inhibit the release of irisin from skeletal muscle in diabetic patients.     

Cardiac alpha- and beta-adrenergic receptor alterations in diabetic cardiomyopathy

Summary The effect of chronic experimental diabetes on the adrenergic receptors in the rat heart was investigated. Diabetes was induced by streptozotocin (65 mg/kg; i.v.) administration, animals were sacrificed 8 weeks later, and positive as well as negative dF/dt values were determined in isolated papillary muscle preparations. Stimulation of the contractile force generation by isoproterenol and methoxamine was attenuated in diabetic preparations. Beta-and alpha-adrenergic receptor bindings were determined in cardiac membranes by employing³H-dihydroalprenolol and³H-dihydroergocryptine respectively. Reduced number of beta- and alpha-receptor binding sites without changes in the affinity constants were observed in diabetic myocardium. Such a decrease in alpha- and beta-receptor density in the heart may account for the depressed contractile responsiveness to adrenergic stimuli in diabetic cardiomyopathy.

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Zusammenfassung Es wurde der Einfluß eines chronischen, experimentellen Diabetes auf die adrenergen Rezeptoren des Rattenherzens untersucht. Diabetes wurde durch Streptozotocin (65 mg/kg i.v.) erzeugt. Die Untersuchungen wurden 8 Wochen nach Verabreichung durchgeführt. Es wurden positive und negative dF/dt-Werte (Geschwindigkeit der Kraftentwicklung) von isolierten Papillarmuskeln bestimmt. Die durch Isoproterenol und Methoxamin gesteigerte Kraftentwicklung war bei Ratten mit Diabetes vermindert. Die Bindungseigenschaften von Beta-und Alpha-rezeptoren in den Membranen des Herzens wurden mit Hilfe von³H-dihydroalprenolol und³H-dihydroergocryptin bestimmt. Im diabetischen Myokard war die Zahl der Beta- und Alpharezeptor-Bindungsstellen vermindert, nicht jedoch die Affinitätskonstanten. Die verringerte Beta- und Alpharezeptoren-Dichte im Herz könnte für die verminderte Ansprechbarkeit der Mechanik auf adrenerge Reize bei der diabetischen Kardiomyopathie verantwortlich sein.

     

FGL2凝血酶原酶在糖尿病大鼠心脏的表达及意义

目的探讨纤维蛋白原样2（FGL2）凝血酶原酶在糖尿病大鼠心脏的表达及其意义。方法实验大鼠随机分为对照（NC）组和2型糖尿病（T2DM）组。HE染色及纤维素染色（改良MSB染色）观察大鼠心脏病理改变,随机计数微血管中微血栓形成的阳性血管数;逆转录pCR法、免疫组织化学法测定FGL2凝血酶原酶在大鼠心脏的表达。结果与NC组相比,FGL2凝血酶原酶在T2DM大鼠的心脏微血管内皮细胞中高表达;T2DM组大鼠心脏微血管内可见微血栓形成;T2DM组大鼠阳性血管数显著高于NC组,且与FGL2凝血酶原酶蛋白表达呈正相关。结论FGL2凝血酶原酶可能通过介导糖尿病心脏微血栓形成而促进糖尿病微血管病变的发生与发展。     

Decrease in calcium-sensing receptor in the progress of diabetic cardiomyopathy

To observe the dynamic expression of calcium-sensing receptor (CaSR) in myocardium of diabetic rats and explore its role in diabetic cardiomyopathy (DCM), 40 male Wistar rats were randomly divided into 4 groups including control, diabetic-4 weeks, diabetic-8 weeks and spermine treatment groups (240 μM of spermine in drinking water). The type 2 Diabetes mellitus (DM) models were established by intraperitoneal injection of streptozotocin (STZ, 30 mg/kg) after high-fat and high-sugar diet for one month. The echocardiographic parameters were measured, cardiac morphology was observed by electron microscope and HE staining. The intracellular calcium concentration ([Ca(2+)](i)) was detected by laser-scanning confocal microscope. Western blot analyzed the expression of CaSR, protein kinase C α(PKC-α) and calcium handling regulators, such as phospholamban (PLN), Ca(2+)-ATPase (SERCA), and ryanodine receptor (RyR). Compared with control group, [Ca(2+)](i) and the expression of CaSR, RyR and SERCA/PLN were decreased, while PKC-α and PLN were significantly increased in a time-dependent manner in diabetic groups. Meanwhile diabetic rats displayed abnormal cardiac structure and systolic and diastolic dysfunction, and spermine (CaSR agonist) could prevent or slow its progression. These results indicate that the CaSR expression of myocardium is reduced in the progress of DCM, and its potential mechanism is related to the impaired intracellular calcium homeostasis.     

Molecular characterization and expression of urotensin II and its receptor in the flounder (Platichthys flesus): a hormone system supporting body fluid homeostasis in euryhaline fish

Urotensin II (UII) is a potent vasoconstrictor in mammals, but the source of circulating UII remains unclear. Investigations of the caudal neurosecretory system (CNSS), considered the major source of UII in fish, alongside target tissue expression of UII receptor (UT), can provide valuable insights into this highly conserved regulatory system. We report UII gene characterization, expression of the first fish UT, and responses to salinity challenge in flounder. The 12-aa UII peptide shares 73% sequence identity with pig and human UII. Flounder UT receptor shares 56.7% identity with rat. Although the CNSS is the major site of UII expression, RT-PCR revealed expression of UII and UT in all tissues tested. Around 30-40% of large CNSS Dahlgren cells expressed UII, alone or in combination with urotensin I and/or corticotrophin releasing hormone. Immunolocalization of UT in osmoregulatory tissues (gill, kidney) was associated with vascular elements. There were no consistent differences in CNSS UII expression or plasma UII between seawater (SW)- and freshwater (FW)-adapted fish, although gill and kidney UT expression was lower in FW animals. After acute transfer from SW to FW, plasma UII and kidney and gill UT expression were reduced, whereas UT expression in kidney was increased after reverse transfer. UII appears to be more important to combat dehydration and salt-loading in SW than the hemodilution faced in FW. Potentially, altered target tissue sensitivity through changes in UT expression, is an important physiological controlling mechanism, not only relevant for migratory fish but also likely conserved in mammals.     

心肌张力蛋白同源物在糖尿病大鼠心肌缺血后处理中的作用

目的:探索糖尿病心肌10号染色体缺失的磷酸酶及张力蛋白同源物(PTEN)蛋白表达的变化及其在糖尿病心肌缺血后处理(IPo)中的作用。方法:选择成年SD大鼠130只,喂养1周后,随机分为糖尿病组(n=90)和非糖尿病组(n=40)。对照组给予等量的0.9%氯化钠溶液。将糖尿病大鼠和非糖尿病大鼠随机分为三组:假手术组(Sham);缺血/再灌注组(IR);缺血后处理组(IPo)。免疫组化方法分析心肌PTEN、磷脂酰肌醇3-激酶(PI3K)、磷酸化蛋白激酶B(P-AKT)的表达;TTC法检测心肌梗死面积;TUNEL法检测心肌细胞凋亡。结果:IPo明显降低了非糖尿病鼠心肌细胞中PTEN的表达,增加了PI-3K和p-Akt的表达,与N+S组相比,N+IR及N+IPo组的凋亡指数明显增高(t=5.102,P=0.029)。但是经后处理干预后N+IPo组的凋亡指数明显低于N+IR组(t=3.997,P=0.041)。与DM+S组相比DM+IR及DM+IPo组的凋亡指数明显增高(t=4.592,P=0.033),但后处理并没有减少糖尿病心肌的凋亡指数。与N+IR相比,N+IPo的梗死面积明显减少(t=5.599,P=0.024)。BPV干预各组的心肌PI-3K及P-Akt的表达明显高于未干预各组。与未经BPV干预的各组相比,被干预各组的心肌凋亡细胞明显减少。心肌梗死面积比较BPV干预的各组心肌梗死明显减少。结论:高血糖导致心肌PTEN高表达是造成PI-3K/Akt信号通路的失活,导致IPo对心肌IRI保护作用丧失的一个重要原因。     

Cloning of the cDNA encoding the urotensin II precursor in frog and human reveals intense expression of the urotensin II gene in motoneurons of the spinal cord

Urotensin II (UII) is a cyclic peptide initially isolated from the caudal neurosecretory system of teleost fish. Subsequently, UII has been characterized from a frog brain extract, indicating that a gene encoding a UII precursor is also present in the genome of a tetrapod. Here, we report the characterization of the cDNAs encoding frog and human UII precursors and the localization of the corresponding mRNAs. In both frog and human, the UII sequence is located at the C-terminal position of the precursor. Human UII is composed of only 11 amino acid residues, while fish and frog UII possess 12 and 13 amino acid residues, respectively. The cyclic region of UII, which is responsible for the biological activity of the peptide, has been fully conserved from fish to human. Northern blot and dot blot analysis revealed that UII precursor mRNAs are found predominantly in the frog and human spinal cord. In situ hybridization studies showed that the UII precursor gene is actively expressed in motoneurons. The present study demonstrates that UII, which has long been regarded as a peptide exclusively produced by the urophysis of teleost fish, is actually present in the brain of amphibians and mammals. The fact that evolutionary pressure has acted to conserve fully the biologically active sequence of UII suggests that the peptide may exert important physiological functions in humans.     

Plasma urotensin II in heart failure

Urotensin II has vasoconstrictive and negative inotropic effects, suggesting a possible role in circulatory regulation and pathophysiology of heart failure. We developed a sensitive specific RIA and measured plasma urotensin II in patients with heart failure and in controls. Plasma urotensin II was higher in heart failure patients (mean 3.9 pmol/L [SD 1.4]; than in controls (1.9 pmol/L [0.9]; p<0.0001). The role of urotensin II in heart failure, however, has yet to be defined.     

Angiotensin II receptor antagonist attenuates expression of aging markers in diabetic mouse heart.

BACKGROUND: Diabetes mellitus is an independent risk factor for heart failure. Diabetes mellitus causes other age-related cardiovascular diseases. We assessed the hypothesis that hearts from diabetic animals are associated with accelerated aging processes. We also examined the effect of an angiotensin II receptor blocker (ARB) on the expression of senescence-associated molecules. METHODS AND RESULTS: We administered an ARB (candesartan 10 mg/kg per day) or saline to diabetic db/db or control db/+ mice. The treatment was started when mice were 10-weeks-old, and continued for 15 weeks. Systolic function was impaired in db/db mice and candesartan improved cardiac function. The amount of phosphorylated Akt and S6 was decreased in saline-treated db/db mice, and candesartan treatment partially preserved phosphorylation. The amount of p21, p27, p53 or Rb was increased in the heart tissue of saline treated db/db mice. Candesartan treatment completely suppressed the increases of p21, p27, p53 and Rb. CONCLUSIONS: An ARB improved cardiac function of diabetic animals, and this was accompanied by decreases of senescence-associated molecules in the myocardium. ARB may be a modality for heart failure patients with diabetes mellitus.     

Vectorcardiographic evaluation of diabetic cardiomyopathy and of its contributing factors

Summary In order to investigate the prevalence of vectorcardiographic bites, expression of small areas of fibrosis, atrophy or degeneration of the myocardium, we studied, using the vectorcardiograms (VCG) of 101 diabetic patients (35 with insulin-dependent and 66 with non-insulin-dependent diabetes mellitus, aged from 25 to 60 years, without hypertension, coronary artery disease, or intraventricular conduction defects) and 228 normal control subjects, matched for age and sex. The prevalence of bites was 38.6% in diabetic patients and 10.0% in the control group (p<0.001). Diabetic patients were also subdivided into groups according to age, sex, metabolic control, risk factors for coronary heart disease, type of diabetes, duration of diabetes and diabetic microangiopathy. No correlation was found between any of the variables investigated nor of a combination of these, and the presence of bites. We conclude that VCG is a sensitive test for cardiac involvement in diabetic patients but that it cannot be used to identify any specific factor able to influence the onset and evolution of this involvement.     

Arrestin-independent internalization and recycling of the urotensin receptor contribute to long-lasting urotensin II-mediated vasoconstriction

Urotensin II (UII), which acts on the G protein-coupled urotensin (UT) receptor, elicits long-lasting vasoconstriction. The role of UT receptor internalization and intracellular trafficking in vasoconstriction has yet not been analyzed. Therefore, UII-mediated contractile responses of aortic ring preparations in wire myography and rat UT (rUT) receptor internalization and intracellular trafficking in binding and imaging analyses were compared. UII elicited a concentration-dependent vasoconstriction of rat aorta (-log EC50, mol/L:9.0+/-0.1). A second application of UII after 30 minutes elicited a reduced contraction (36+/-4% of the initial response), but when applied after 60 minutes elicited a full contraction. In internalization experiments with radioactive labeled VII ((125)I-UII), approximately 70% of rUT receptors expressed on the cell surface of human embryonic kidney 293 cells were sequestered within 30 minutes (half life [t(h)]: 5.6+/-0.2 minutes), but recycled quantitatively within 60 minutes (t(h) 31.9+/-2.6 minutes). UII-bound rUT receptors were sorted to early and recycling endosomes, as evidenced by colocalization of rUT receptors with the early endosomal antigen and the transferrin receptor. Real-time imaging with a newly developed fluorescent UII (Cy3-UII) revealed that rUT receptors recruited arrestin3 green fluorescent protein to the plasma membrane. Arrestin3 was not required for the endocytosis of the rUT receptor, however, as internalization of Cy3-UII was not altered in mouse embryonic fibroblasts lacking endogenous arrestin2/arrestin3 expression. The data demonstrate that the rUT receptor internalizes arrestin independently and recycles quantitatively. The continuous externalization of rUT receptors provides the basis for repetitive and lasting UII-mediated vasoconstriction.     

Role of urotensin II in patients on dialysis

Urotensin II is a potent vasoconstrictor, which also has some vasodilatory properties. We investigated its expression in various tissues and in the plasma of patients with renal dysfunction. Plasma concentrations of urotensin II-like immunoreactivity were 2-fold higher in patients not on dialysis and 3-fold higher in those on haemodialysis thanin healthy individuals. Messenger RNA encoding theurotensin II precursor and the urotensin II receptor precursor were expressed in various human tissues. The peptidemight act as an important regulator in the cardiovascularand renal systems. Urotensin II antagonists could, therefore, be useful in the treatment of diseases affecting theseorgans.