Direct Anti-inflammatory Effects Research Articles

Association of Novel Antihyperglycemic Drugs Versus Metformin With a Decrease in Asthma Exacerbations

BackgroundSimilar to metformin, dipeptidyl peptidase-4 inhibitors (DPP-4 Is), glucagon-like peptidase 1 receptor agonists (GLP-1 RAs), and sodium glucose co-transporter-2 inhibitors (SGLT-2 Is) may improve control of asthma owing to their multiple potential mechanisms, including differential improvements in glycemic control, direct anti-inflammatory effects, and systemic changes in metabolism. ObjectiveTo investigate whether these novel antihyperglycemic drugs were associated with fewer asthma exacerbations compared with metformin in patients with asthma comorbid with type 2 diabetes. MethodsUsing a Japanese national administrative database, we constructed three active comparators—new user cohorts of 137,173 patients with a history of asthma starting the novel antihyperglycemic drugs and metformin between 2014 and 2022. Patient characteristics were balanced using overlap propensity score weighting. The primary outcome was the first exacerbation requiring systemic corticosteroids, and the secondary outcomes included the number of exacerbations requiring systemic corticosteroids. ResultsDPP-4 Is and GLP-1 RAs were associated with a higher incidence of exacerbations requiring systemic corticosteroids compared with metformin (DPP-4 Is: 18.2 vs. 17.4 per 100 person-years; hazard ratio 1.09, 1.05 to 1.14; GLP-1 RAs: 24.9 vs. 19.0 per 100 person-years; 1.14, 1.01 to 1.28). In contrast, the incidence of exacerbations requiring systemic corticosteroids was similar between the SGLT-2 Is and metformin groups (17.3 vs. 18.1 per 100 person-years; hazard ratio 1.00, 0.97 to 1.03). While DPP-4 Is and GLP-1 RAs were associated with more exacerbations requiring systemic corticosteroids, SGLT-2 Is were associated with slightly fewer exacerbations requiring systemic corticosteroids (53.7 vs. 56.6 per 100 person-years; rate ratio 0.95, 0.91 to 0.99). ConclusionsWhile DPP-4 Is and GLP-1 RAs were associated with poorer control of asthma compared with metformin, SGLT-2 Is offered asthma control comparable to that of metformin.

Metoprolol disrupts inflammatory response of human cardiomyocytes via β-arrestin2 biased agonism and NF-κB signaling modulation

AimsRecent evidence supports non-class cardioprotective effects of metoprolol against neutrophil-mediated ischemia-reperfusion injury during exacerbated inflammation. Whether metoprolol exerts direct anti-inflammatory effect on cardiomyocytes is unknown. Accordingly, we aimed to investigate the direct anti-inflammatory effects of metoprolol in a cellular model of human induced pluripotent stem cell-derived cardiomyocytes (hiCMs) and to explore the role of β-arrestin2 (β-ARR2) biased agonism signaling pathway. Methods and resultshiCMs were treated with TNF-α for 24 h, followed by 4-hour treatment with metoprolol or esmolol. Electrical response of hiCMs to β1-selective blockade was assessed by microelectrode arrays technology. The effect on inflammatory and adhesion molecule expression was evaluated in wild-type and β-ARR2 silenced hiCMs. To silence β-ARR2 expression, hiCMs were transfected with a specific small interfering RNA targeting β-ARR2 mRNA and preventing its translation.TNF-α stimulation boosted the expression of IκB, NF-κB, IL1β, IL6, and VCAM1 in hiCMs. TNF-α-treated hiCMs showed similar physiological responses to metoprolol and esmolol, with no difference in field potential duration and beat period recorded. Adding metoprolol significantly decreased inflammatory response patterns in wild-type hiCMs by dampening TNF-α induced expression of NF-κB, IL1β, and IL6, but not in β-ARR2-knockout hiCMs. A similar response was not observed in presence of β1-selective blockade with esmolol. ConclusionsMetoprolol exerts a non-class direct anti-inflammatory effect on hi-CMs. β1-selective blockade with metoprolol disrupts inflammatory responses induced by TNF-α and induces significant inhibition of NF-κB signaling cascade via β-ARR2 biased agonism. If confirmed at clinical level, metoprolol could be tested and repurposed to treat cardiac inflammatory disorders.

Pneumokinetic therapy with glucocorticoids and 5 FU in the treatment of hypertrophic scars

Corticosteroid injections, ointments, and patches are quite effective for hypertrophic scars. In addition to their direct anti-inflammatory effects, steroids also act by causing vasoconstriction. Current evidence suggests that 5‑FU is an alternative to glucocorticoids for the treatment of hypertrophic scars. According to research, 5‑FU can significantly improve the appearance of pathological scars and reduce the likelihood of relapse. Recently published studies have shown the effectiveness of laser therapies, including pulsed dye laser, fractional CO2‑laser, copper bromide laser, and UVA1 phototherapy. Purpose: to evaluate the effectiveness of pneumokinetic administration of drugs. Material and methods. The study group included 34 patients with hypertrophic scars who underwent 2 procedures of pneumokinetic therapy with glucocorticoids and 5‑FU, with an interval of 2 months. After another 1 month, laser therapy was performed with a neodymium laser (1064 nm). Efficiency control was carried out using DUB SkinScanner, a device for ultrasound examination and determination of morphofunctional parameters of the skin. Results. The study group showed pronounced positive dynamics. The thickness of the epidermis increased by 25.4 %; echodensity decreased by 17.8 %. The thickness of the dermis and its echodensity are more indicative; in relation to these parameters, the following dynamics were noted: a decrease in the thickness of the dermis by 48.0 % and an increase in echoicity by 46.6 %. Ultrasound data revealed a higher efficiency of the combined use of pneumokinetic therapy with glucocorticoids and 5‑FU with sequential laser therapy (neodymium laser at 1064 nm). Conclusion: The combined use of pneumokinetic therapy with glucocorticoids and 5‑FU with sequential laser therapy (neodymium laser at 1064 nm) is a highly effective method of treating hypertrophic scars and can be recommended for use in practical healthcare.

Remibrutinib (LOU064) inhibits neuroinflammation driven by B cells and myeloid cells in preclinical models of multiple sclerosis

BackgroundBruton’s tyrosine kinase (BTK) is a key signaling node in B cell receptor (BCR) and Fc receptor (FcR) signaling. BTK inhibitors (BTKi) are an emerging oral treatment option for patients suffering from multiple sclerosis (MS). Remibrutinib (LOU064) is a potent, highly selective covalent BTKi with a promising preclinical and clinical profile for MS and other autoimmune or autoallergic indications.MethodsThe efficacy and mechanism of action of remibrutinib was assessed in two different experimental autoimmune encephalomyelitis (EAE) mouse models for MS. The impact of remibrutinib on B cell-driven EAE pathology was determined after immunization with human myelin oligodendrocyte glycoprotein (HuMOG). The efficacy on myeloid cell and microglia driven neuroinflammation was determined in the RatMOG EAE. In addition, we assessed the relationship of efficacy to BTK occupancy in tissue, ex vivo T cell response, as well as single cell RNA-sequencing (scRNA-seq) in brain and spinal cord tissue.ResultsRemibrutinib inhibited B cell-dependent HuMOG EAE in dose-dependent manner and strongly reduced neurological symptoms. At the efficacious oral dose of 30 mg/kg, remibrutinib showed strong BTK occupancy in the peripheral immune organs and in the brain of EAE mice. Ex vivo MOG-specific T cell recall response was reduced, but not polyclonal T cell response, indicating absence of non-specific T cell inhibition. Remibrutinib also inhibited RatMOG EAE, suggesting that myeloid cell and microglia inhibition contribute to its efficacy in EAE. Remibrutinib did not reduce B cells, total Ig levels nor MOG-specific antibody response. In brain and spinal cord tissue a clear anti-inflammatory effect in microglia was detected by scRNA-seq. Finally, remibrutinib showed potent inhibition of in vitro immune complex-driven inflammatory response in human microglia.ConclusionRemibrutinib inhibited EAE models by a two-pronged mechanism based on inhibition of pathogenic B cell autoreactivity, as well as direct anti-inflammatory effects in microglia. Remibrutinib showed efficacy in both models in absence of direct B cell depletion, broad T cell inhibition or reduction of total Ig levels. These findings support the view that remibrutinib may represent a novel treatment option for patients with MS.

Lobeglitazone inhibits LPS-induced NLRP3 inflammasome activation and inflammation in the liver.

Liver inflammation is a common feature of chronic liver disease and is often associated with increased exposure of the liver to lipopolysaccharide (LPS). Kupffer cells (KCs) are macrophages in the liver and produce various cytokines. Activation of KCs through the NLRP3 inflammasome pathway leads to release of proinflammatory cytokines and induces hepatocyte injury and hepatic stellate cell (HSC) activation. Lobeglitazone is a peroxisome proliferator-activated receptor gamma ligand and a type of thiazolidinedione that elicits anti-inflammatory effects. However, there is no clear evidence that it has direct anti-inflammatory effects in the liver. This study showed that lobeglitazone reduces LPS-induced NLPR3 inflammasome activation and production of proinflammatory cytokines in primary KCs and hepatocytes. Cytokines secreted by activated KCs increased hepatocyte inflammation and HSC activation, and lobeglitazone inhibited these responses. In addition, lobeglitazone suppressed liver fibrosis by inhibiting LPS-induced transforming growth factor (TGF)-β secretion and TGF-β-induced CTGF expression. The inhibitory effect of lobeglitazone on inflammasome activation was associated with suppression of liver fibrosis. These results suggest that lobeglitazone may be a treatment option for inflammation and fibrosis in the liver.

Omega-3 Polyunsaturated Fatty Acids and Their Anti-Oxidant, Anti-Inflammatory and Neuroprotective Effects in Diabetic Retinopathy: A Narrative Review.

Diabetic retinopathy (DR) is a common microvascular complication of type 2 Diabetes Mellitus (T2DM) that can have vision-threatening consequences, particularly if it advances to the proliferative stage and is left untreated. Owing to the central role of hyperglycemia-induced oxidative stress, multiple anti-oxidants have been investigated for their therapeutic value. However, there is a lack of substantial data to support the use of any of the compounds tested so far. Omega-3 polyunsaturated fatty acids (PUFAs), namely docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), have received much acclaim for their positive impact on cardiovascular health outcomes. The anti-oxidative, anti-inflammatory and neuroprotective properties of PUFAs also make them promising therapeutic and preventive agents for DR. The current evidence is derived mainly from in vitro and animal studies and provides some insight into the underlying mechanisms involved. These fatty acids are capable of direct anti-oxidative and anti-inflammatory effects. They also concomitantly promote intrinsic defense mechanisms and recovery, particularly of photoreceptor neurons. Hence, dietary supplementation with PUFAs, mainly from marine sources, can halt and reverse the retinal damage seen in DR. Furthermore, clinical trials have reported improved vision and quality of life in DR patients after supplementation. However, a major limitation of these trials is the use of nutraceutical formulations in which omega-3 PUFAs are combined with other anti-oxidant compounds, thereby preventing the evaluation of omega-3 as standalone treatment. Although the results of experimental studies to date have been promising, more clinical trials are required to determine the full extent of benefits in patients with DR.

Brain-derived neurotrophic factor (BDNF) has direct anti-inflammatory effects on microglia.

Microglia are the primary immunocompetent cells that protect the brain from environmental stressors, but can also be driven to release pro-inflammatory cytokines and induce a cytotoxic environment. Brain-derived neurotrophic factor (BDNF) is important for the regulation of plasticity, synapse formation, and general neuronal health. Yet, little is known about how BDNF impacts microglial activity. We hypothesized that BDNF would have a direct modulatory effect on primary cortical (Postnatal Day 1-3: P1-3) microglia and (Embryonic Day 16: E16) neuronal cultures in the context of a bacterial endotoxin. To this end, we found that a BDNF treatment following LPS-induced inflammation had a marked anti-inflammatory effect, reversing the release of both IL-6 and TNF-α in cortical primary microglia. This modulatory effect was transferrable to cortical primary neurons, such that LPS-activated microglial media was able produce an inflammatory effect when added to a separate neuronal culture, and again, BDNF priming attenuated this effect. BDNF also reversed the overall cytotoxic impact of LPS exposure in microglia. We speculate that BDNF can directly play a role in regulating microglia state and hence, influence microglia-neuron interactions.

Stimulation of Angiotensin II Type 2 Receptor Modulates Pro-Inflammatory Response in Microglia and Macrophages: Therapeutic Implications for the Treatment of Stroke.

Sustained microglial activation contributes to the development of post-stroke cognitive impairment (PSCI). Compound 21 (C21), an angiotensin II type 2 receptor agonist, has shown some neurovascular protection after stroke. This study aimed to investigate the direct anti-inflammatory effects of C21 on macrophages, as well as brain innate immune cells. Murine microglial cell line (C8-B4) and RAW 264.7 macrophages were exposed to lipopolysaccharide (LPS) and co-treated with C21. Pro-inflammatory mediators were assessed via RT-qPCR and ELISA. Cellular reactive oxygen species (ROS) were evaluated via CellROXGreen staining, and nitrate production was assessed using Griess assay. C21 suppressed LPS-induced inflammation and ROS generation in both cells. In microglia, C21 blunted LPS-induced mRNA expression of IL-1β, IL-12b, COX-1, iNOS, and IL-6. A similar pattern was observed in macrophages, where C21 suppressed LPS-induced IL-1β, TNF-α, and CXCL1 expression. These anti-inflammatory effects in microglia and macrophages were associated with increased neuroprotective gene expression, including GDNF and BDNF, in a dose-dependent manner. Our findings suggest a protective effect of C21 against the inflammatory response, in both macrophages and microglia, via suppression of the release of pro-inflammatory cytokines/chemokines and the generation of ROS while stimulating the production of neurotrophic factors.

Trial of Tauroursodeoxycholic Acid Supplementation in Patients with Progressive MS (S16.007)

<h3>Objective:</h3> The main aim of this trial was to assess safety and tolerability of tauroursodeoxycholic acid (TUDCA) supplementation in patients with progressive multiple sclerosis (pwPMS). Secondary objectives were to assess the effects on serum BA profile and clinical outcomes. <h3>Background:</h3> We have previously identified changes in the metabolome, specifically in bile acid (BA) metabolism in MS. We demonstrated that BA receptors are present in MS lesions, and TUDCA supplementation ameliorated disease in an animal model of MS and had direct anti-inflammatory effects on astrocytes and microglia. Trials in other neurodegenerative diseases have shown that TUDCA may be neuroprotective. The effects of TUDCA supplementation on MS are unknown but could provide a pathway for developing new therapies in PMS. <h3>Design/Methods:</h3> In this randomized, double-blinded trial, pwPMS were enrolled and randomized 1:1 to either receive TUDCA (1 g twice daily) or placebo for 16 weeks. Clinical visits occurred at 0, 8, and 16 weeks. Primary outcomes were safety and tolerability of TUDCA. Secondary outcomes included changes in BA serum levels and clinical outcomes. <h3>Results:</h3> Of the 59 participants enrolled, 47 (80%) completed ≥2 visits and were included in the analysis (21 in placebo arm, 26 in TUDCA arm). The number of participants reporting adverse events was not significantly different between groups (10 in TUDCA arm vs 7 in placebo arm; p=0.72). The TUDCA group demonstrated significant increases over time in levels of TUDCA (p&lt;0.001), glycoursodeoxycholic acid (p&lt;0.001), ursodeoxycholic acid (p&lt;0.001), and lithocholic acid (p=0.02). There was a significant improvement in MSFC (p=0.03), while no change was noted in EDSS and MSQOL-54 mental or physical composites in the TUDCA arm; no significant changes were noted in these outcomes in the placebo arm. <h3>Conclusions:</h3> TUDCA supplementation was safe and tolerable in pwPMS, and led to significant increases in levels of several bioactive secondary BAs and improvement in MSFC score. <b>Disclosure:</b> The institution of Dr. Harrison has received research support from National MS Society. Dr. Ladakis has nothing to disclose. Mr. Hwang has nothing to disclose. Dr. Sotirchos has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Ad Scientiam. Dr. Sotirchos has received personal compensation in the range of $5,000-$9,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Viela Bio. Dr. Sotirchos has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Genentech. Dr. Sotirchos has received personal compensation in the range of $10,000-$49,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Alexion. The institution of Dr. Sotirchos has received research support from National Institutes of Health. The institution of Dr. Sotirchos has received research support from National Multiple Sclerosis Society. The institution of Dr. Sotirchos has received research support from Sumaira Foundation. The institution of Dr. Sotirchos has received research support from Genentech. Dr. Sotirchos has received personal compensation in the range of $500-$4,999 for serving as a Ad Hoc Reviewer with National Institutes of Health. Dr. Calabresi has received personal compensation in the range of $5,000-$9,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Biogen. Dr. Calabresi has received personal compensation in the range of $10,000-$49,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Disarm Therapeutics. Dr. Calabresi has received personal compensation in the range of $500-$4,999 for serving as an Editor, Associate Editor, or Editorial Advisory Board Member for Journal of Clinical Investigation. The institution of Dr. Calabresi has received research support from Biogen. The institution of Dr. Calabresi has received research support from Annexon. Dr. Calabresi has received publishing royalties from a publication relating to health care. Dr. Calabresi has received personal compensation in the range of $500-$4,999 for serving as a Study Section Member with NIH. Dr. Calabresi has received personal compensation in the range of $500-$4,999 for serving as a Grant reveiwer with Myelin Repair Foundation. Dr. Calabresi has received personal compensation in the range of $500-$4,999 for serving as a Speaker for CME with NYAS. Dr. Calabresi has received personal compensation in the range of $500-$4,999 for serving as a Speaker with Academic CME. Dr. Calabresi has a non-compensated relationship as a Consultant with Apellis that is relevant to AAN interests or activities. The institution of Dr. Bhargava has received research support from EMD Serono. The institution of Dr. Bhargava has received research support from Amylyx pharmaceuticals. The institution of Dr. Bhargava has received research support from Genentech. The institution of Dr. Bhargava has received research support from GSK.

Protective Effect of Novel Lactobacillus plantarum KC3 Isolated from Fermented Kimchi on Gut and Respiratory Disorders.

Probiotics have been shown to possess anti-inflammatory effects in the gut by directly reducing the production of pro-inflammatory cytokines and by secreting anti-inflammatory molecules. However, their systemic anti-inflammatory effects have not been thoroughly investigated. In this study, we aimed to develop probiotics that have efficacy in both intestinal and lung inflammation. Lactobacillus plantarum KC3 (KC3), which was isolated from kimchi, was selected as a pre-candidate based on its inhibitory effects on the production of pro-inflammatory cytokines in vitro. To further validate the effectiveness of KC3, we used ear edema, DSS-induced colitis, and ambient particulate-matter-induced lung inflammation models. First, KC3 exhibited direct anti-inflammatory effects on intestinal cells with the inhibition of IL-1β and TNF-α production. Additionally, KC3 treatment alleviated ear edema and DSS-induced colic inflammation, improving colon length and increasing the number of regulatory T cells. Beyond its local intestinal anti-inflammatory activity, KC3 inhibited pro-inflammatory cytokines in the bronchoalveolar fluid and prevented neutrophil infiltration in the lungs. These results suggest that KC3 could be a potential functional ingredient with respiratory protective effects against air-pollutant-derived inflammation, as well as for the treatment of local gut disorders.

Possibilities of combined intranasal drugs in the treatment of patients with rhinosinusitis

Rhinosinusitis – an inflammatory process in the nasal cavity and paranasal sinuses is a global problem of practical medicine, which remains relevant for many years. The most common cause of acute rhinosinusitis is a viral infection, only in 0.2–5 % of patients the disease is further complicated by the formation of a bacterial focus of inflammation. Treatment of patients withARS involves the use of etiotropic, pathogenetic and symptomatic therapy. National clinical guidelines consider the use of systemic and topical etiotropic drugs, intranasal glucocorticosteroids, topical decongestants, irrigation- elimination procedures, mucoactive therapy, standardized herbal medicine, drainage of the paranasal sinuses, antihistamines (with concomitant allergies) and some other activities. Local administration of drugs in patients with rhinosinusitis provides targeted delivery of the active substance to the pathological focus and the creation of its required concentration, the virtual absence of systemic action due to low absorption, as well as a favorable softening, moisturizing, enveloping effect of auxiliary components. One of these drugs is the combined nasal spray containing tuaminoheptane sulfate and acetylcysteine. The effectiveness and safety of the use of this combination has been demonstrated by the results of a fairly large amount of research. The mild and sufficiently long vasoconstrictive effect of tuaminoheptane sulfate, the direct mucolytic, antioxidant and anti-inflammatory effects of acetylcysteine and the high safety of this combination determine its therapeutic value in children from 6 years of age and adult patients with rhinitis and rhinosinusitis.

Impact of High-Density Lipoproteins on Sepsis.

Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. Here, we review the impact of high-density lipoproteins (HDL) on sepsis from the perspective of biochemistry and pathophysiology, epidemiological research, and intervention studies in animals. Pathogen lipid moieties are major ligands for innate immunity receptors, such as toll-like receptors. The binding of pathogen-associated lipids to lipoproteins leads to sequestration, neutralization, and inactivation of their pro-inflammatory effects. Lipoproteins constitute an arm of the innate immune system. Pathogen-associated lipids can be removed from the body via the reverse lipopolysaccharide transport pathway in which HDL play a key role. Independent of the capacity for sequestration, the direct anti-inflammatory effects of HDL may counteract the development of sepsis. Mendelian randomization research using genetic variants associated with HDL cholesterol as an instrumental variable was consistent with a probable causal relationship between increased HDL cholesterol levels and decreased risk of infectious hospitalizations. Low HDL cholesterol independently predicts an adverse prognosis in sepsis both in observational epidemiology and in Mendelian randomization studies. Several HDL-associated enzymes, including phospholipid transfer protein (PLTP) and cholesterol ester transfer protein (CETP), undergo profound changes during sepsis. Potential HDL-directed interventions for treatment of sepsis include apolipoprotein A-I-based therapies, recombinant PLTP, and CETP inhibition.

Effect of exercise-based cardiac rehabilitation on the ischemic threshold in patients with high-risk ischemic heart disease

Abstract Introduction Coronary artery disease is a complex condition and is not always resolved after an interventional or surgical procedure, due to the remaining obstructions in the coronary tree or the severity of the atherosclerosis, leaving anginal symptoms, ischemic threshold (IU) in the exercise, decreased cardiorespiratory fitness, and poor quality of life (QoL). Exercise-based cardiac rehabilitation programs (Ex-CRP) usually develop ischemic preconditioning mechanisms thanks to their direct anti-inflammatory, antioxidant, antithrombotic and coronary neovascularization effects, thus improving coronary reserve flow. We hypothesize that an Ex-CRP is capable of favorably impacting the UI in these patients. Therefore, the objective of this study is to evaluate the effects of an Ex-CRP on the UI in patients with high-risk ischemic heart disease. Methods Quasi-experimental study with 121 patients with ischemic threshold due to high-risk ischemic heart disease (with perfusion defects in nuclear medicine, complex coronary anatomy or partially revascularized) and presence of ischemic threshold in the exercise test. An Ex-CRP of 4–6 weeks duration was performed, consisting of supervised and monitored training concurrent with 30-minute aerobic exercise sessions at 70% of heart rate reserve, 3 times per week, supplemented with resistance exercise. and an educational understanding program for the control of risk factors and nutritional and psycho-emotional care. At the end of the program, a stress test was performed to compare the post-intervention effects and ischemic threshold was defined as the presence of angina or electrocardiographic changes of ischemia with negative ST-segment depression in 2 consecutive beats more than 80 ms from the J point. and in more than two contiguous leads. Results The 121 patients with ischemic threshold were 61±10 years old and 75% were male. On the other hand, 29.7% had a diagnosis of heart failure and an average LVEF of 53.1±12.9%. After Ex-CRP, an improvement was found in calculated VO2p (24.7±7.8 vs 34.23±7.5; p=0.000), METs at maximum effort from 7.06±2.15 to 9.78±1.14 (p&amp;lt;0.0001) and submaximal load in watts from 44±21.32 to 61±24.49; p&amp;lt;0.0001. 40.5% of the patients who were admitted with UI resolved it (absence of post-intervention ischemic diagnosis) after Ex-CRP and the remaining percentage with UI (59.5%) displaced it by 2.62 METs (p&amp;lt;0.0001). With this, the cardiovascular risk calculated by the Duke Score also decreased from −7 to −2 (p&amp;lt;0.0001), and the myocardial efficiency index improved from 8 to 6 (p&amp;lt;0.0001), strength (79±15 to 92±8%; p&amp;lt;0.0001) and QoL (65±17 vs 82±12; p&amp;lt;0.0001). Conclusions Ex-CRP had a significant effect on the resolution or displacement of the UI in patients with high-risk ischemic heart disease. In addition, it improved muscle strength, quality of life, myocardial efficiency, and reduced the cardiovascular risk index calculated by the Duke score. Funding Acknowledgement Type of funding sources: Private company. Main funding source(s): Centro de Estudios e Investigaciόn FISICOL. Cúcuta, ColombiaCardioFit. CDMX, México

Esaxerenone Blocks Vascular Endothelial Inflammation Through SGK1.

Chronic low-grade inflammation and excess mineralocorticoid receptor (MR) activation are well-known pathological conditions of metabolic syndrome (MetS). To elucidate the crosstalk between inflammation and MR signaling, we focused on serum/glucocorticoid-regulated kinase 1 (SGK1) in vascular endothelial cells. We treated human aortic endothelial cells (HAECs) with esaxerenone (ESX), a novel nonsteroidal highly selective MR antagonist, or spironolactone (SPL), a classic competitive MR antagonist, followed by stimulation with tumor necrosis factor (TNF)-α. ESX at therapeutic concentrations attenuated the long-term induction of TNF-α-stimulated inflammatory molecules in HAEC, whereas SPL had only a minor effect at 10 μM. We found long-term TNF-α-stimulated induction of SGK1 mRNA and protein levels in HAEC and that ESX pretreatment significantly decreased SGK1 mRNA and protein levels at both the basal and the TNF-α-stimulated conditions, whereas SPL had no effect on SGK1 mRNA and protein levels. In addition, the TNF-α-induced nuclear factor kappa-light-chain-enhancer of activated B cell activity was suppressed by the treatment with ESX, and it was abrogated by SGK1 overexpression. These results indicated that ESX has direct anti-inflammatory effects in HAEC via the blocking of long-term TNF-α-induced SGK1 activation and that SGK1 could be a key molecule linking cytokine-induced vascular chronic inflammation and MR activation.

FRI157 - Direct anti-inflammatory and anti-fibrotic effects of a novel long-acting Glucagon/GIP/GLP-1 triple agonist, HM15211, in thioacetamide-induced mouse model of liver injury and fibrosis

FRI157 - Direct anti-inflammatory and anti-fibrotic effects of a novel long-acting Glucagon/GIP/GLP-1 triple agonist, HM15211, in thioacetamide-induced mouse model of liver injury and fibrosis

735-P: Direct Anti-Inflammatory and Antifibrotic Effects of a Novel Long-Acting Glucagon/GIP/GLP-1 Triple Agonist, HM15211, in Thioacetamide-Induced Mouse Model of Liver Injury and Fibrosis

HM152 is a novel long-acting triple agonist consisting of Glucagon/GIP/GLP-1 triple agonist conjugated to human IgG FC fragment via short PEG linker. Previously, therapeutic benefits of HM152 were demonstrated in diet-induced animal models of NASH and fibrosis. Here, direct anti-inflammatory and -fibrotic effects of HM152 were further evaluated in TAA (thioacetamide) -induced liver injury and fibrosis mouse, and investigate underlying mechanism. To induce liver injury and fibrosis, gradually increased dose of TAA was injected to mouse for 12 weeks, and HM152 was administered during last weeks. Interestingly, HM152 treatment significantly reduced hepatic hydroxyproline (-51% vs. Veh, p&amp;lt;0.01) , Sirius red positive area (-65% vs. Veh, p&amp;lt;0.001) , and fibrosis score (0.7 for HM152 vs. 3.0 for Veh, p&amp;lt;0.001) . Considering baseline fibrosis score at week 2 (1.0) , HM152 could confer both potential reversal effect on pre-existing fibrosis and prevention effect on fibrogenesis. Consistently, expression of hepatic marker genes for fibrosis (i.e. collagen-1α1) and inflammation (i.e. F4/80, TNF-α) were significantly reduced in HM152 group. Furthermore, multiplex cytokine analysis revealed that HM152 treatment was associated with robust reduction across pro-inflammatory cytokines including TNF-α, IL-1β, and IL-6. Significant reduction in blood level of liver enzymes was also confirmed. Mechanistically, PMA/LPS-induced pro-inflammatory cytokine secretion of THP-1 cell was significantly attenuated by HM15211. TGF-β-induced collagen production was also reduced in LX2 cell. In conclusion, HM152 markedly improved liver inflammation and fibrosis in TAA mice, and related mechanism was elaborated by in vitro studies. Thus, HM152 could be a novel therapeutic option for fibrosis due to NASH. Human study is ongoing to assess the clinical relevance of these findings. Disclosure J.Kim: None. S.Lee: n/a. I.Choi: None. J.Lee: None. Y.Kim: None. S.Lee: None. H.Kwon: Employee; Hanmi Pharm. Co., Ltd. E.Park: Employee; Hanmi Pharm. Co., Ltd. J.Choi: Employee; Hanmi Pharm. Co., Ltd. S.Bae: None. D.Kim: None.

874-P: Potential Anti-Inflammatory Effects of Canagliflozin in Tumour Necrosis Factor-α and Interferon-ϒ-Stimulated Endothelial Cells

Background: This study investigates the potential direct anti-inflammatory effects of the SGLT2 inhibitors (canagliflozin, C; dapagliflozin D; or empagliflozin, E) on cultured endothelial cells. Methods: Human umbilical vein endothelial cells (HUVECs) or Human cardio microvascular endothelial cells (HCMECs) were pre-incubated in the presence or absence of C (0.5μM, 1μM, and 10μM) , D (0.5μM, 1μM, and 10μM) and E (0.5μM, 1μM, and 10μM) for 1 hr and NaCl (150 mM) for 24 hr. The cells were then stimulated with TNF-α (ng/mL) and IFN-Υ (ng/mL) for 24 hrs. Expression of IL-6 and was analysed using Quantikine ELISA kits. The expression of SGLT2 mRNA was assessed using RT-PCR in cDNA prepared from two independent cultures of HUVECs and HCMECs using cDNA from human kidney cortex as a positive control. Results: TNF-α and IFN-Υ and significantly induced expression of IL-6 in both HUVECs and HCMECs compared to controls (p = &amp;lt;0.0001) . The effects were significantly inhibited by canagliflozin 0.1μM, 1μM, and 10μM (p=0.0099, p&amp;lt; 0.0001, and p&amp;lt;0.0001, respectively) in HUVECs. In HCMECs, C, (0.5μM, 1μM, and 10μM) significantly inhibit the TNF-α and IFN-Υ IL-6 secretion (p&amp;lt; 0.0001, p&amp;lt; 0.0001, and p&amp;lt;0.0001, respectively) . No significant effect of D and E was observed. High NaCl had no influence on C (0.5μM, 1μM, and 10μM) inhibition of TNF-α and IFN-Υ induced of IL-6 secretion, and high salt alone had no influence on the secretion of IL-6 secretion. SGLT2 mRNA levels were below the detection level in HUVECs and HCMECs alone and on high glucose and TNF-α and IFN-Υ conditions. Conclusion: This result indicates that SGLT2-I directly inhibits endothelial pro-inflammatory cytokines secretions. This striking anti-inflammatory of the SGLT2 inhibitor class merits further mechanistic research. Disclosure A.S.Alshnbari: None. R.N.Khan: None. I.R.Idris: None.

Effects of Glycyrrhizic Acid in Licorice on Prolongation of Murine Cardiac Allograft Survival

Recent evidence has pointed to the promising benefits of using specific immunosuppressive herbal compounds to prolong transplant allograft survival. In this study, we investigated the effects of glycyrrhizic acid (GA), a major component of licorice, in a model of murine heart transplantation. CBA (H2k) mice were transplanted with a fully-MHC mismatched C57BL/6 (H2b) heart allograft and subsequently received daily intraperitoneal administration of normal saline or 0.02, 0.2, or 2.0 mg/d of GA for 7 consecutive days. Untreated CBA recipients, with a median survival time (MST) of 7 days, and groups receiving 0.02mg/d (MST, 8 days) or 0.2mg/d (MST, 9 days) of GA acutely rejected C57BL/6 cardiac allografts. But mice treated with 2.0 mg/d of GA demonstrated significant prolongation of allografts (MST, 23 days). Histologic studies showed that cardiac allografts from GA-treated CBA recipients had preserved graft and vessel structure. Moreover, flow cytometric study showed that the percentage of CD4+CD25+Foxp3+ cell (regulatory T cell [Treg]) populations were increased in GA-treated CBA recipients. In a mixed leukocyte culture, splenocytes from GA-treated mice demonstrated suppressed allo-proliferation, in which interleukin (IL)-2 and interferon gamma production were downregulated and IL-10 secretion was upregulated. In conclusion, GA may be a novel promising therapeutic agent to prolong cardiac allograft survival through direct anti-inflammatory effects and induction of Treg populations.

Effects of Colchicine in a Rat Model of Diet-Induced Hyperlipidemia.

Inflammation and hyperlipidemia play an essential role in the pathophysiology of endothelial dysfunction as well as atherosclerotic plaque formation, progression and rupture. Colchicine has direct anti-inflammatory effects by inhibiting multiple inflammatory signaling pathways. The purpose of our study was to evaluate colchicine activity in an animal model of hyperlipidemia induced by diet. A total of 24 male rats (wild type, WT) were divided into three groups: group one fed with a basic diet (BD) (WT + BD, n = 8), group two fed with a high-fat diet (HFD) (WT + HFD, n = 8)), and group three which received HFD plus drug treatment (colchicine, 0.5 mg/kg, i.p., daily administration). Total cholesterol, LDL-, HDL-cholesterol and triglycerides were determined. In addition, plasma transaminases, inflammation of oxidative stress markers, were measured. Tissue samples were evaluated using hematoxylin-eosin and red oil stain. At the end of the study, rats presented increased serum lipid levels, high oxidative stress and pro-inflammatory markers. The aortic histopathological section revealed that HFD induced signs of endothelial dysfunction. Colchicine treatment significantly resolved and normalized these alterations. Moreover, colchicine did not influence NAFLD activity score but significantly increased ALT and AST levels, suggesting that colchicine amplified the hepatocellular injury produced by the diet. Colchicine reduces plasma lipid levels, oxidative stress and inflammation markers and leads to more favorable histopathologic vascular and cardiac results. However, the adverse effects of colchicine could represent an obstacle to its safe use.

Dapagliflozin exerts anti-inflammatory effects via inhibition of LPS-induced TLR-4 overexpression and NF-κB activation in human endothelial cells and differentiated macrophages

Evidence has demonstrated that a new class of anti-diabetic drugs, sodium-glucose co-transporter 2 (SGLT2) inhibitors, could exert beneficial effects on atherosclerotic complications of diabetes. Atherosclerosis is widely accepted as an inflammatory disease. Therefore, we aimed to assess the direct anti-inflammatory effects of SGLT2 inhibitors dapagliflozin (DAPA) on two cell types involved in the process of atherogenesis. Human umbilical vein endothelial cells (HUVECs) and macrophages were exposed to DAPA and lipopolysaccharide (LPS 20 ng/mL) for 24 h under normal (5.5 mmol/L, NG) or high glucose (25 mmol/L, HG) conditions. Then, levels of TLR-4/p–NF–κB, inflammatory cytokines, inflammation-related miR-146a and miR-155 as well as alteration in the ratio of M1/M2 macrophage polarization was assessed. DAPA (0.5 μM) could significantly attenuate LPS-induced TLR-4 overexpression (23.9% and 33.1% under NG and HG conditions in HUVECs and 53.3% and 52.4% under NG and HG states in macrophages, respectively). NF-κB p65 phosphorylation was also significantly decreased to 30.1% under NG condition in HUVECs and 51.9% and 34.5% under NG and HG states in macrophages by 0.5 μM DAPA. Moreover, DAPA elevated expression levels of anti-inflammatory miR-146a, while values of miR-155 decreased in those cells. DAPA also caused a shift from inflammatory M1 macrophages toward M2-dominant macrophages. These data suggest that regardless of glucose concentrations, DAPA could exert direct anti-inflammatory effects, at least partly, by inhibiting the expression of TLR-4 and activation of NF-κB along with the secretion of pro-inflammatory mediators.