Analgesia mediated by soluble epoxide hydrolase inhibitors is dependent on cAMP

Pain is a major health concern even though numerous analgesic agents are available. Side effects and lack of wide-spectrum efficacy of current drugs justify efforts to better understand pain mechanisms. Stabilization of natural epoxy-fatty acids (EFAs) through inhibition of the soluble epoxide hydrolase (sEH) reduces pain. However, in the absence of an underlying painful state, inhibition of sEH is ineffective. Surprisingly, a pain-mediating second messenger, cAMP, interacts with natural EFAs and regulates the analgesic activity of sEH inhibitors. Concurrent inhibition of sEH and phosphodiesterase (PDE) dramatically reduced acute pain in rodents. Our findings demonstrate a mechanism of action of cAMP and EFAs in the pathophysiology of pain. Furthermore, we demonstrate that inhibition of various PDE isozymes, including PDE4, lead to significant increases in EFA levels through a mechanism independent of sEH, suggesting that the efficacy of commercial PDE inhibitors could result in part from increasing EFAs. The cross-talk between the two major pathways-one mediated by cAMP and the other by EFAs-paves the way to new approaches to understand and control pain.     

Experimental colitis in mice is attenuated by changes in the levels of endocannabinoid metabolites induced by selective inhibition of fatty acid amide hydrolase (FAAH)

Background and aimsPharmacological treatment and/or maintenance of remission in inflammatory bowel diseases (IBD) is currently one of the biggest challenge in the field of gastroenterology. Available therapies are mostly limited to overcoming the symptoms, but not the cause of the disease. Recently, the endocannabinoid system has been proposed as a novel target in the treatment of IBD. Here we aimed to assess the anti-inflammatory action of the novel fatty acid amide hydrolase (FAAH) inhibitor PF-3845 and its effect on the endocannabinoid and related lipid metabolism during the course of experimental colitis.MethodsWe used two models of experimental colitis in mice (TNBS- and DSS-induced) and additionally, we employed LC/MS/MS spectrometry to determine the changes in biolipid levels in the mouse colon during inflammation.ResultsWe showed that the FAAH inhibitor PF-3845 reduced experimental TNBS-induced colitis in mice and its anti-inflammatory action is associated with altering the levels of selected biolipids (arachidonic and oleic acid derivatives, prostaglandins and biolipids containing glycine in the mouse colon).ConclusionsWe show that FAAH is a promising pharmacological target and the FAAH-dependent biolipids play a major role in colitis. Our results highlight and promote therapeutic strategy based on targeting FAAH-dependent metabolic pathways in order to alleviate intestinal inflammation.     

抑制可溶性环氧物酶对局灶性脑缺血再灌注大鼠的神经保护作用

目的 探讨可溶性环氧物酶(soluble epoxide hydrolase,sEH)抑制剂12-(3-金刚烷-1-基脲基)-十二烷酸[12-(3-adamant an-1-yl-ureido)-dodec-anoic acid,AUDA]对局灶性脑缺血再灌注大鼠的神经保护作用和机制.方法 60只雄性Sprague-Dawley大鼠随机分为假手术组、生理盐水对照组以及小剂量(0.157 ml/kg)、中剂量(0.235 ml/kg)和大剂量(0.314 ml/kg) AUDA处理组(每组12只),各组随机取4只大鼠分别用于梗死体积、细胞凋亡和p-Akt免疫组织化学检测.线栓法建立大脑中动脉闭塞再灌注模型.各AUDA处理组和生理盐水对照组均在再灌注前分别经腹腔给予相应剂量的AUDA或等体积生理盐水.再灌注24 h时进行神经功能缺损评分.2,3,5-氯化三苯基四氮唑染色法检测脑梗死体积.原位缺口末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)检测梗死周围区脑组织细胞凋亡.免疫组织化学法检测梗死周围区脑组织p-Akt表达.结果 TTC染色显示,假手术组未见梗死.生理盐水对照组以及小剂量、中剂量和大剂量AUDA组梗死体积分别为(254.146±25.481)、(212.679±7.514)、(150.188±33.997)和(99.563±3.415) mm3,存在显著性差异(F=39.637,P=0.000).各剂量AUDA组均显著性小于对照组(P均=0.000).中剂量AUDA组显著性小于小剂量AUDA组(P=0.002),而大剂量AUDA也显著性小于小剂量AUDA组(P =0.000)和中剂量AUDA组(P=0.006).TUNEL染色法显示,假手术组仅可见少量凋亡细胞[(6.400±1.477)个/高倍视野].生理盐水对照组以及小剂量、中剂量和大剂量AUDA组凋亡细胞数量分别为(57.550±13.067)、(47.030±8.423)、(34.530±4.393)和(26.400±2.683)个/高倍视野,各剂量AUDA组显著性少于生理盐水对照组(P均＜0.01),中剂量和大剂量AUDA组显著性少于小剂量AUDA组(P均＜0.01),大剂量AUDA组也显著性少于中剂量AUDA组(P＜0.01).免疫组织化学显示,假手术组仅可见少量p-Akt阳性细胞[(3.325±1.438)个/高倍视野],生理盐水对照组以及小剂量、中剂量和大剂量AUDA组p-Akt阳性细胞数量分别为(9.450 ±2.531)、(16.400 ±3.865)、(22.875±7.974)和(29.300±3.203)个/高倍视野,各剂量AUDA组显著性多于生理盐水对照组(P均＜0.01),中剂量和大剂量AUDA组显著性多于小剂量AUDA组(P均＜0.01),大剂量AUDA组也显著性多于中剂量AUDA组(P＜0.01).结论 抑制sEH可能通过上调PI3K/Akt通路减少梗死周围区神经元凋亡和缩小梗死体积,对局灶性脑缺血再灌注大鼠具有神经保护作用.     

Inhibition of soluble epoxide hydrolase reduces food intake and increases metabolic rate in obese mice

Background and aimsThis study evaluated the responses to soluble epoxide hydrolase (s-EH) inhibition, an essential enzyme in the metabolism of arachidonic acid, on food intake, body weight and metabolic parameters in mice fed a high fat-high fructose diet (HFD) for 10 weeks.Methods and resultsAfter 5 weeks of HFD, mice were divided into two groups: 1) s-EH inhibitor (AR9281, 200 mg/kg/day by gavage twice daily), and 2) vehicle (0.3 ml per gavage). Food intake, body weight, oxygen consumption (VO₂), carbon dioxide production (VCO₂), respiratory quotient (RQ), and motor activity were measured weekly for more 5 weeks. HFD increased body weight (37 ± 1 vs. 26 ± 1 g), and plasma of glucose (316 ± 8 vs. 188 ± 27 mg/dl), insulin (62.1 ± 8.1 vs. 15.5 ± 5.0 μU/ml), and leptin levels (39.4 ± 3.6 vs. 7.5 ± 0.1 ng/ml) while reducing VO₂, VCO₂ and motor activity. s-EH inhibition for 5 weeks decreased caloric intake by ～32% and increased VO₂ by ～17% (42.8 ± 1.4 vs. 50.2 ± 1.5 ml/kg/min) leading to significant weight loss. Inhibition of s-EHi also caused significant reductions in plasma leptin levels and visceral fat content. Uncoupling protein 1 (UCP1) content in brown adipose tissue was also elevated by ～50% during s-EH inhibition compared to vehicle treatment.ConclusionThese results suggest that s-EH inhibition with AR9281 promotes weight loss by reducing appetite and increasing metabolic rate, and that increased UCP1 content may contribute to the increase in energy expenditure.     

Gene deficiency and pharmacological inhibition of soluble epoxide hydrolase confers resilience to repeated social defeat stress

Depression is a severe and chronic psychiatric disease, affecting 350 million subjects worldwide. Although multiple antidepressants have been used in the treatment of depressive symptoms, their beneficial effects are limited. The soluble epoxide hydrolase (sEH) plays a key role in the inflammation that is involved in depression. Thus, we examined here the role of sEH in depression. In both inflammation and social defeat stress models of depression, a potent sEH inhibitor, TPPU, displayed rapid antidepressant effects. Expression of sEH protein in the brain from chronically stressed (susceptible) mice was higher than of control mice. Furthermore, expression of sEH protein in postmortem brain samples of patients with psychiatric diseases, including depression, bipolar disorder, and schizophrenia, was higher than controls. This finding suggests that increased sEH levels might be involved in the pathogenesis of certain psychiatric diseases. In support of this hypothesis, pretreatment with TPPU prevented the onset of depression-like behaviors after inflammation or repeated social defeat stress. Moreover, sEH KO mice did not show depression-like behavior after repeated social defeat stress, suggesting stress resilience. The sEH KO mice showed increased brain-derived neurotrophic factor (BDNF) and phosphorylation of its receptor TrkB in the prefrontal cortex, hippocampus, but not nucleus accumbens, suggesting that increased BDNF-TrkB signaling in the prefrontal cortex and hippocampus confer stress resilience. All of these findings suggest that sEH plays a key role in the pathophysiology of depression, and that epoxy fatty acids, their mimics, as well as sEH inhibitors could be potential therapeutic or prophylactic drugs for depression.Depression is the most severe and debilitating of the psychiatric illnesses. The World Health Organization estimates that more than 350 million individuals of all ages suffer from depression (1). Almost one million lives are lost annually because of suicide, which translates to 3,000 deaths daily (1). Although antidepressants are generally effective in the treatment of depression, it can still take weeks before patients feel the full antidepressant effects. However, approximately two-thirds of depressed patients fail to respond fully to pharmacotherapy. Furthermore, there is a high rate of relapse, and depressed patients have a high risk of committing suicide (2–4).Accumulating evidence suggests that inflammation plays a central role in the pathophysiology of depression (5–9). Meta-analyses showed higher blood levels of proinflammatory cytokines, such as tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6), in drug-free depressed patients compared with healthy controls (10–13). Studies using postmortem brain samples showed elevated gene expression of proinflammatory cytokines in the frontal cortex of people with a history of depression (14, 15). Taking these data together, we find that it is likely that both peripheral and central inflammations are associated with depression and that antiinflammatory drugs, such as cyclooxygenase inhibitors, could ameliorate depressive symptoms in depressed patients (16, 17).Epoxyeicosatrienoic acids (EETs), which are produced from arachidonic acid by the action of cytochrome P450s, have potent antiinflammatory actions. These mediators are broken down into the corresponding diols by soluble epoxide hydrolase (sEH), and inhibition of sEH enhances the beneficial effects of EETs (18–21). It is also reported that sEH inhibitors have potent antiinflammatory effects in a number of animal models (18–20, 22, 23). Although sEH has been associated with the onset of anorexia nervosa (24), the role of sEH in the pathophysiology of depression has not been studied to date.The purpose of this study was to examine the role of sEH in the pathophysiology of depression using a potent sEH inhibitor and sEH knockout (KO) mice. Furthermore, we examined the role of brain-derived neurotrophic factor (BDNF) and its receptor TrkB signaling in selected brain regions, because BDNF-TrkB signaling plays a key role in the pathophysiology of depression (25–30).     

Protection of mice against lethal endotoxemia by lipid X is mediated through inhibition of neutrophil function

The effects of lipid X and 3-aza-lipid X on in vitro neutrophil function were related to their ability to inhibit the toxicity of endotoxin in galactosamine-sensitized mice. In vitro, lipid X and 3-aza-lipid X (100 ng/ml) blocked completely endotoxin (100 ng/ml)-enhanced neutrophil aggregation, superoxide anion generation, and release of beta-glucuronidase in response to a chemotactic tripeptide, f-met-leu-phe (10(-7) M). In vivo, lipid X at 250 micrograms/mouse (but not 3-aza-lipid X at a similar dose) protected groups of 10 mice from an otherwise lethal dose of endotoxin in galactosamine-sensitized mice when it was administered IV 4 hr or 2 hr before endotoxin challenge. The minimum effective dose of lipid X that could protect 50% of the challenged mice was calculated to be 715 micrograms/kg. However, lipid X failed to suppress neutrophil infiltration into the lungs. The ability of lipid X to inhibit endotoxin-induced neutrophil responses and to protect against lethal endotoxemia may be due to induction of early phase tolerance to endotoxin by the compound.     

Protection from endotoxic shock in mice by pharmacologic inhibition of phosphatidic acid.

Certain phosphatidic/plasmanic/plasmenic acid (PA) species function as lipid intermediates in cell activation and may function directly as intracellular signaling molecules. PA can also be dephosphorylated to 1,2-diradyl-sn-glycerol by phosphatidate phosphohydrolase. Treatment of various cell types, including murine P388 monocytic leukemia cells, with bacterial lipopolysaccharide rapidly stimulates large increases in PA and PA-derived diradylglycerol. Pentoxifylline, 1-(5-oxohexyl)-3,7-dimethylxanthine, inhibits lipopolysaccharide-stimulated formation of PA in P388 cells at high concentrations (IC50 = 500 microM). Lisofylline [1-(5R-hydroxyhexyl)-3,7-dimethylxanthine] is a unique metabolite of pentoxifylline in humans and is > 800-fold more active as an inhibitor of PA formation than pentoxifylline (IC50 = 0.6 microM). Lisofylline does not inhibit lipopolysaccharide-induced activation of phosphatidylinositol-specific phospholipase C and generation of phosphatidylinositol-derived diradylglycerol. Lisofylline but not pentoxifylline protects BALB/c mice from endotoxin lethality when administered 4 hr after lipopolysaccharide. This protective effect is independent of either agent's effect on suppression of plasma tumor necrosis factor alpha. These data suggest that inhibitors of PA formation may have significant clinical potential in the treatment of sepsis and septic shock.     

Soluble epoxide hydrolase regulates hematopoietic progenitor cell function via generation of fatty acid diols

Fatty acid epoxides are important lipid signaling molecules involved in the regulation of vascular tone and homeostasis. Tissue and plasma levels of these mediators are determined by the activity of cytochrome P450 epoxygenases and the soluble epoxide hydrolase (sEH), and targeting the latter is an effective way of manipulating epoxide levels in vivo. We investigated the role of the sEH in regulating the mobilization and proliferation of progenitor cells with vasculogenic/reparative potential. Our studies revealed that sEH down-regulation/inhibition impaired the development of the caudal vein plexus in zebrafish, and decreased the numbers of lmo2/cmyb-positive progenitor cells therein. In mice sEH inactivation attenuated progenitor cell proliferation (spleen colony formation), but the sEH products 12,13-dihydroxyoctadecenoic acid (12,13-DiHOME) and 11,12- dihydroxyeicosatrienoic acid stimulated canonical Wnt signaling and rescued the effects of sEH inhibition. In murine bone marrow, the epoxide/diol content increased during G-CSF-induced progenitor cell expansion and mobilization, and both mobilization and spleen colony formation were reduced in sEH(-/-) mice. Similarly, sEH(-/-) mice showed impaired functional recovery following hindlimb ischemia, which was rescued following either the restoration of bone marrow sEH activity or treatment with 12,13-DiHOME. Thus, sEH activity is required for optimal progenitor cell proliferation, whereas long-term sEH inhibition is detrimental to progenitor cell proliferation, mobilization, and vascular repair.     

可溶性环氧化物水解酶抑制剂对脂肪细胞胆固醇流出的影响

目的 观察可溶性环氧化合物水解酶抑制剂(sEHi)tAUCB对脂肪细胞胆固醇流出的影响,并探讨其机制.方法 测定脂肪细胞过氧化物酶体增殖物激活受体γ(PPARγ)及三磷酸腺苷结合盒转运体A1(ABCA1)mRNA及蛋白的表达,检测细胞内胆固醇流出.结果 tAUCB呈剂量依赖性促进载脂蛋白(Apo)A1介导的胆固醇流出,1、10、50、100 μmol/L浓度的tAUCB干预后,胆固醇流出率分别为(5.93±0.66)%,(7.40±0.43)%,(8.30±0.34)%和(9.77±0.42)%.加入10～100μmol/L tAUCB干预组与空白组(5.67±0.17)%比较,胆固醇流出差异有统计学意义(P＜0.05).同时发现随着tAUCB干预浓度的增加,脂肪细胞ABCA1、PPARy mRNA及蛋白的表达也呈剂量依赖性地升高,而GW9662明显抑制tAUCB对脂肪细胞胆固醇流出及ABCA1和PPARγ表达的促进作用.结论 tAUCB可通过上调PPARy的表达,促进脂肪细胞Apo A1-ABCA1通路加速细胞内胆固醇流出,抑制脂肪细胞内胆固醇过度蓄积.

Abstract：

Objective To observe the effects of soluble epoxide hydrolase inhibitors tAUCB on cholesterol efflux in adipocytes. Methods 3T3-L1 preadipocytes were induced to differentiation and maturation. Cells were stimilated with 100μg/L LPS after starved for 24 hours, then tAUCB in various concentrations(1 ,10,50,100 μmol/L)were added for 24 h, or incubated with the peroxisome proliferator activated receptor gamma (PPARy) antagonist GW9662 (5 μmol/L).0μmol/L tAUCB treated group was taken as empty control. After then, the mRNA expression of PPARγ and adenosine triphosphate binding cassette transporter Al (ABCA1) in cells were determined via realtime-PCR, the amounts of protein expression of PPARγand ABCA1 in cells were detected by Western blot, the efflux rates of 3H-cholesterol in cells were detected by means of liquid scintillation counter. Results tAUCB could dose-dependently increase the apolipoprotein A1 (apoA1)-mediated cholesterol efflux in adipocytes. After stimulated by 1, 10,50,100 μmol/L tAUCB, cholesterol efflux rates were (5.93±0.66) %, (7.40 ± 0. 43) %, (8. 30 ±0. 34)% ,(9.77±0.42)% respectively, there were significant difference after treated by 10-100 μmol/L tAUCB compared with control(5.67±0.17)%(P＜0.05). With the concentration of tAUCB increased,ABCA1, PPAR mRNA and protein expression were also dose-dependently up-regulated. GW9662 could significantly inhibit the effects of tAUCB, and then reduce the cholesterol efflux and the expression of PPARγ and ABCA1 in adipocytes. Conclusions tAUCB could up-regulate PPARγ expression in adipocytes, and promote the cholesterol efflux of adipocytes via apoA1-ABCA1 pathway, which might decrease the cellular cholesterol accumulation in adipocytes.     

Attenuation of tobacco smoke-induced lung inflammation by treatment with a soluble epoxide hydrolase inhibitor

Changes in the lungs due to smoking include inflammation, epithelial damage, and remodeling of the airways. Airway inflammation is likely to play a critical role in the genesis and progression of tobacco smoke-induced airway disease. Soluble epoxide hydrolase (sEH) is involved in the metabolism of endogenous chemical mediators that play an important role in inflammation. Epoxyeicosatrienoic acids (EETs) have demonstrated antiinflammatory properties, and hydrolysis of these epoxides by sEH is known to diminish this activity. To examine whether acute tobacco smoke-induced inflammation could be reduced by a sEH inhibitor, 12-(3-adamantane-1-yl-ureido)-dodecanoic acid n-butyl ester was given by daily s.c. injection to spontaneously hypertensive rats exposed to filtered air or tobacco smoke for a period of 3 days (6 h/day). Acute exposure to tobacco smoke significantly increased by 3.2-fold (P <0.05) the number of cells recovered by bronchoalveolar lavage. The sEH inhibitor significantly decreased total bronchoalveolar lavage cell number by 37% in tobacco smoke-exposed rats with significant reductions noted in neutrophils, alveolar macrophages, and lymphocytes. A combination of sEH inhibitor and EETs was more significant in its ability to further reduce tobacco smoke-induced inflammation compared with the sEH inhibitor alone. The sEH inhibitor led to a shift in some plasma epoxides and diols that are consistent with the hypothetical action of these compounds. We conclude that an sEH inhibitor, in the presence or absence of EETs, can attenuate, in part, inflammation associated with acute exposure to tobacco smoke.     

Protection of susceptible BALB/c mice from challenge with Leishmania major by nucleoside hydrolase, a soluble exo-antigen of Leishmania

Leishmania major culture-derived, soluble, exogenous antigens have been shown to be a source of vaccine targets for the parasite. We have previously reported that L. major culture-derived, soluble, exogenous antigens can immunize BALB/c mice against challenge with L. major. However, the molecule(s) involved in this protection was not known. We describe the potential of one component of soluble exogenous antigens (recombinant nucleoside hydrolase) to vaccinate mice against challenge with L. major. We found that recombinant nucleoside hydrolase vaccinated BALB/c mice against a subsequent challenge with L. major. Protection was manifested by a significant decrease in lesion size (as much as a 30-fold reduction) and parasite burden (as much as a 71-fold reduction). Protection was achieved whether recombinant nucleoside hydrolase was administered to mice in the presence or absence of adjuvant (interleukin-12). Finally, protection was accompanied by an increase in interferon-gamma production but a decrease in interleukin-10 production by vaccinated animals in response to challenge with L. major.     

Variations in the human soluble epoxide hydrolase gene and recurrence of atrial fibrillation after catheter ablation

Background

Single nucleotide polymorphisms (SNPs) of EPHX2 alter sEH activity and are associated with increased [rs41507953 (K55R)] or reduced [rs751141 (R287Q)] cardiovascular risk via modulation of fibrosis, inflammation or cardiac ion channels. This indicates an effect on development and therapy response of AF. This study tested the hypothesis that variations in the EPHX2 gene encoding human soluble epoxide hydrolase (sEH) are associated with atrial fibrillation (AF) and recurrence of atrial fibrillation after catheter ablation.

Methods and results

A total of 218 consecutive patients who underwent catheter ablation for drug refractory AF and 268 controls were included. Two SNPs, rs41507953 and rs751141, were genotyped by direct sequencing. In the ablation group, holter recordings 3, 12 and 24 months after ablation were used to detect AF recurrence. No significant association of the SNPs and AF at baseline was detected. In the ablation group, recurrence of AF occurred in 20% of the patients 12 months after ablation and in 35% 24 months after ablation. The presence of the rs751141 polymorphism significantly increased the risk of AF recurrence 12 months (odds ratio [OR]: 3.2, 95% confidence interval [CI]: 1.237 to 8.276, p = 0.016) and 24 months (OR: 6.076, 95% CI: 2.244 to 16.451, p < 0.0001) after catheter ablation.

Conclusions

The presence of rs751141 polymorphism is associated with a significantly increased risk of AF recurrence after catheter ablation. These results point to stratification of catheter ablation by genotype and differential use of sEH-inhibitory drugs in the future.     

Cytochrome P450 epoxygenases, soluble epoxide hydrolase, and the regulation of cardiovascular inflammation

The cytochrome P450 (CYP) epoxygenase enzymes CYP2J and CYP2C catalyze the epoxidation of arachidonic acid to epoxyeicosatrienoic acids (EETs), which are rapidly hydrolyzed to dihydroxyeicosatrienoic acids (DHETs) by soluble epoxide hydrolase (sEH). It is well-established that CYP epoxygenase-derived EETs possess potent vasodilatory effects; however, the cellular effects of EETs and their regulation of various inflammatory processes have become increasingly appreciated in recent years, suggesting that the role of this pathway in the cardiovascular system extends beyond the maintenance of vascular tone. In particular, CYP epoxygenase-derived EETs inhibit endothelial activation and leukocyte adhesion via attenuation of nuclear factor-kappaB activation, inhibit hemostasis, protect against myocardial ischemia-reperfusion injury, and promote endothelial cell survival via modulation of multiple cell signaling pathways. Thus, the CYP epoxygenase pathway is an emerging target for pharmacological manipulation to enhance the cardiovascular protective effects of EETs. This review will focus on the role of the CYP epoxygenase pathway in the regulation of cardiovascular inflammation and (1) describe the functional impact of CYP epoxygenase-derived EET biosynthesis and sEH-mediated EET hydrolysis on key inflammatory process in the cardiovascular system, (2) discuss the potential relevance of this pathway to pathogenesis and treatment of cardiovascular disease, and (3) identify areas for future research.     

Unique mechanistic insights into the beneficial effects of soluble epoxide hydrolase inhibitors in the prevention of cardiac fibrosis

Tissue fibrosis represents one of the largest groups of diseases for which there are very few effective therapies. In the heart, myocardial infarction (MI) resulting in the loss of cardiac myocytes can culminate in adverse cardiac remodeling leading to eventual heart failure. Adverse cardiac remodeling includes myocyte hypertrophy, fibrosis, and electrical remodeling. We have previously demonstrated the beneficial effects of several potent soluble epoxide hydrolase inhibitors (sEHIs) in different models of cardiac hypertrophy and failure. Here, we directly determine the molecular mechanisms underlying the beneficial effects of sEHIs in cardiac remodeling post-MI. Treatment with a potent sEHI, 1-trifluoromethoxyphenyl-3-(1-propionylpiperidine-4-yl)urea (TPPU), which was started 1 wk post-MI in a murine model, results in a significant improvement in cardiac function. Importantly, treatment with TPPU results in a decrease in cardiac fibrosis as quantified using histological and immunostaining techniques. Moreover, single-cell–based assays demonstrate that treatment with TPPU results in a significant decrease not only in the percentages but also the proliferative capacity of different populations of cardiac fibroblasts as well as a reduction in the migration of fibroblasts into the heart from the bone marrow. Our study provides evidence for a possible unique therapeutic strategy to reduce cardiac fibrosis and improve cardiac function post-MI.     

Health benefits of Mediterranean diet in nonalcoholic fatty liver disease

Introduction: Nonalcoholic fatty liver disease (NAFLD) is the most common liver disease worldwide. Insulin resistance and obesity-related inflammatory status, associated with genetic, dietary, and lifestyle factors, are involved in its pathogenesis. There is no consensus concerning the pharmacological treatment of NAFLD. However, the international guidelines agree to define a dietetic nutritional management to achieve weight loss, as an essential component of any therapeutic strategy.

Areas covered: An overview on the beneficial effects of the Mediterranean diet in the prevention and treatment of NAFLD.

Expert commentary: On the basis of its components, the literature reports the beneficial effects of the Mediterranean diet in preventing major chronic diseases, including obesity, diabetes, cardiovascular diseases, and some forms of cancers. In recent years, a growing body of evidence has supported the idea that the Mediterranean diet, associated with physical activity and cognitive behavior therapy, may be the reference nutritional profile for the prevention and the treatment of NAFLD patients.