Expression Of GPR43 Research Articles

Changes of Short-Chain Fatty Acids and Their Receptors in an Obese Rat Model After Sleeve Gastrectomy.

Short-chain fatty acids (SCFAs) and gut microbiota have health-related effects and are associated with a wide range of disorders. However, the changes of SCFAs and their receptors after sleeve gastrectomy (SG) remain unclear. This study aimed to examine changes of SCFAs and their receptors after SG in an obese rat model. Thirty obese Sprague-Dawley rats eating a high-energy diet for 6weeks were divided into three groups: sham-operated (SO) control, pair-fed (PF) control, and SG group. Six weeks after the surgery, metabolic parameters, SCFA levels in the blood and stool, mRNA and protein expression of SCFA receptors in the ileum and epididymal fat, and gut microbiota were examined. Metabolic parameters in the SG group were significantly improved compared with the SO group. Acetic acid levels in the blood and stool were significantly higher in the SG group than the PF group. The butyric acid level in the stool was also significantly higher in the SG group than in the PF group. In the ileum and epididymal fat, mRNA and protein expression of GPR41 was significantly higher in the SG group than in the other two groups, and mRNA and protein expression of GPR43 was significantly higher in the SG group than in the PF group. Increases in the genera Enterococcus, Lactobacillus, Lactococcus, and Clostridium were observed in the stool after SG. SG may activate SCFA pathways through a change in gut microbiota.

Agonism of GPR120 Prevented High Glucose-Induced Apoptosis of Retinal Endothelial Cells through Inhibiting NLRP3 Inflammasome

GPR120 has been reported to ameliorate inflammation in diabetes and diabetic complications. In this study, GW9508, the GPR120 agonist, was utilized in human retinal microvascular endothelial cells (HRMECs) exposed to high glucose (HG) to investigate the involvement of GPR120 in cellular viability and apoptosis as well as the association with the NLRP3 inflammasome. The expression of GPR120 in HRMECs cultured under HG was firstly detected by Western blotting. HRMECs were then assigned to the normal control, GW9508, HG, and HG + GW9508 groups. The expression of the NLRP3 inflammasome consists of NLRP3, ASC, and caspase-1 and was detected by Western blotting and the downstream IL-1β and IL-18 by ELISA. The cellular viability and apoptosis of HRMECs were detected by CCK-8 and flow cytometry, respectively. The expressions of apoptosis-related proteins Bax and Bcl-2 were detected by Western blotting. Finally, nonspecific siRNA (NS) or GPR120 siRNA (siGPR120) was transfected to the cells, followed by stimulation with or without GW9508 or HG, and the expression of NLRP3, ASC, and caspase-1 were detected by Western blotting in these groups. GPR120 is expressed in HRMECs, and HG can reduce its expression in a time-dependent manner. GW9508 can attenuate inflammation by reducing the expression of NLRP3, ASC, caspase-1, IL-1β, and IL-18 under HG. GW9508 rescues the viability of HRMCs and reduces cell apoptosis by preventing an increase in Bax expression and the reduction in Bcl-2 expression. Additionally, knockdown of GPR120 by siRNA weakened the effects of GW9508 on NLRP3 inflammasome expression. Activation of GPR120 protects retinal vascular endothelial cells from HG through inhibiting NLRP3 inflammasome. Thus, GPR120 might be a potential therapeutic target to reduce retinal endothelial damage in diabetic retinopathy.

Clinicopathologic and genomic characteristics of mucinous gastric adenocarcinoma.

Mucinous gastric adenocarcinoma (MGC) is a rare but distinctive histologic subtype of gastric cancer (GC). The clinico-pathologic and genomic characteristics of MGC have not been well evaluated. We collected individual data from five cohorts targeting the microsatellite instability (MSI) of GC (n = 5089) to evaluate the clinico-pathologic characteristics of MGC. In addition, public genomic databases were used for genomic analysis. The characteristics of MGC were compared with those of non-mucinous GC (NMGC). MGC (n = 158, 3.1%) showed distinctive characteristics in terms of age, sex, and TNM stage compared to NMGC (n = 4931). MGC was frequently associated with MSI-high (OR: 2.24, 95% confidence interval [CI] 1.44-3.40, p < 0.001), while mutually exclusive to the Epstein-Barr virus type. The prognosis of MGC was better than that of NMGC (adj.HR: 0.731, 95% CI 0.556-0.962, p = 0.025). There was no clear benefit from postoperative chemotherapy in MGC. TP53 was the main driver mutation in the MGC without recurrent variants. MGC was related to high expression of GPR120 and B3GNT6 and moderate regulation of epithelial-mesenchymal transition (EMT)-up signature with a high EMT-down signature, and those characteristics was related to favorable prognosis of GC (log-rank p = 0.044, p < 0.001, p < 0.001, respectively). MSI-H of MGC was associated with low cancer-associate fibroblasts but high CD274 (PD-L1) expression compared to microsatellite stable MGC, suggesting that immune checkpoint inhibitors may be useful for the MSI-H of MGC. MGC could be a surrogate for performing MSI but not the EBV test in GC. Further, its genetic characteristics lead to a favorable prognosis for MGC.

Protective Roles of Sodium Butyrate in Lipopolysaccharide-Induced Bovine Ruminal Epithelial Cells by Activating G Protein-Coupled Receptors 41

This study aimed to evaluate whether sodium butyrate (SB) attenuates the ruminal response to LPS-stimulated inflammation by activating GPR41 in bovine rumen epithelial cells (BRECs). We examined the SB regulation of GPR41 and its impact on LPS-induced inflammation using GPR41 knockdown BRECs. The LPS-induced BRECs showed increases in the expression of genes related to pro-inflammation and decreases in the expression of genes related to tight junction proteins; these were attenuated by pretreatment with SB. Compared with that in LPS-stimulated BRECs, the ratio of phosphorylated NF-κB (p65 subunit) to NF-κB (p65 subunit) and the ratio of phosphorylated IκBα to IκBα were suppressed with SB pretreatment. The LSB group abated LPS-induced apoptosis and decreased the expression of Bax, Caspase 3, and Caspase 9 mRNA relative to the LPS group. In addition, the LSB group had a lower proportion of cells in the G0–G1 phase and a higher proportion of cells in the S phase than the LPS group. The mRNA expression of ACAT1 and BDH1 genes related to volatile fatty acid (VFA) metabolism were upregulated in the LSB group compared to those in LPS-induced BRECs. In addition, pretreatment with SB promoted the gene expression of GPR41 in the LPS-induced BRECs. Interestingly, SB pretreatment protected BRECs but not GPR41KD BRECs. Our results suggest that SB pretreatment protects against the changes in BRECs LPS-induced inflammatory response by activating GPR41.

GPR43 signaling in lung eosinophils suppresses neutrophilic airway inflammation in asthma

Abstract Asthma is caused by chronic inflammation in airways. It typically manifests type 2 immune activation and eosinophilia, while more severe, steroid-resistant asthma involves additional activation of Th17 cells and neutrophils. Even though pathogenic roles of eosinophils in asthma have been extensively studied, regulatory mechanisms curving down their overactivation are not well known. We found that eosinophils in asthmatic lung highly upregulate expression of GPR43, a receptor for short-chain fatty acid mediating various anti-inflammatory responses. In eosinophil-specific GPR43-deficient (GPR43ΔEOS) mice, asthma induction by Ova/alum or house dust mite extracts resulted in increased activation of Th17 cells and neutrophils in the lung compared to wild type mice. Consequently, GPR43ΔEOS mice showed more severe lung pathology and disease phenotype. Moreover, we identified a distinct Siglec-F+ neutrophil subset is enriched in GPR43ΔEOS mice by scRNA-seq analysis of the asthmatic lungs. Siglec-F+ neutrophils were recently shown to mediate Th17 cell activation and aggravation of asthma in vivo. We also found that Siglec-F+ neutrophils promote in vitro Th17 differentiation. Co-culture experiments of lung eosinophils and bone marrow neutrophils revealed that GPR43-deficient eosinophils directly mediate conversion of Siglec-F-neutrophils into Siglec-F+ neutrophils via GM-CSF secretion. Combined, our results demonstrate that GPR43 signaling in lung eosinophils prevents generation of Siglec-F+ neutrophils and Th17 cell activation, thereby suppressing severe asthma pathology.

Low Intestinal IL22 Associates With Increased Transplant-Related Mortality After Allogeneic Stem Cell Transplantation.

The role of IL-22 in adult patients undergoing allogeneic stem cell transplantation (SCT) is of major interest since animal studies showed a protective and regenerative effect of IL-22 in graft versus host disease (GvHD). However, no clinical data exist on the tissue expression. Here we demonstrate that patients not suffering from transplant-related mortality (TRM) show significantly upregulated IL22 expression during histological and clinical GI-GvHD (p = 0.048 and p = 0.022, respectively). In contrast, in GvHD patients suffering from TRM, IL22 was significantly lower (p = 0.007). Accordingly, lower IL22 was associated with a higher probability of TRM in survival analysis (p = 0.005). In a multivariable competing risk Cox regression analysis, low IL22 was identified as an independent risk factor for TRM (p = 0.007, hazard ratio 2.72, 95% CI 1.32 to 5.61). The expression of IL22 seemed to be microbiota dependent as broad-spectrum antibiotics significantly diminished IL22 expression (p = 0.019). Furthermore, IL22 expression significantly correlated with G-protein coupled receptor (GPR)43 (r = 0.263, p = 0.015) and GPR41 expression (r = 0.284, p = 0.009). In conclusion, our findings reveal an essential role of IL22 for the prognosis of patients undergoing allogeneic SCT.

Exogenous butyrate regulates lipid metabolism through GPR41-ERK-AMPK pathway in rabbits

Diets containing higher levels of fat can lead to obesity, which is potentially harmful to health. Endogenous butyric acid is produced by intestinal microbial fermentation and participates in lipid metabolism. However, there is less butyric acid produced in the body, butyric acid has a shorter half-life in the blood. We used intraperitoneal injection of sodium butyrate to study the mechanism of butyrate involved in body lipid metabolism. Triglycerides in adipose tissue and liver were significantly reduced by butyrate. The content of triglyceride in plasma in butyrate group was also significantly decreased. In adipose tissue, butyrate up-regulated gene expression of hormone-sensitive lipase (HSL), carnitine palmitoyl transferase (CPT)1 and CPT2, and increased protein expression of G protein-coupled receptor (GPR)41, phosphorylated extracellular-signal-regulated kinase (P-ERK), adenosine 5’-monophosphate (AMP)-activated protein kinase (AMPK) and P-AMPK. Gene expression of lipoprotein lipase (LPL), differentiation-dependent factor 1 (ADD1) and fatty acid synthase (FAS) was down-regulated. In liver, butyrate up-regulated protein expression of GPR41, ERK, P-ERK, P-AMPK. Gene expression of ADD1 and FAS was down-regulated. In muscle tissue, butyrate significantly up-regulated the expression of gene LPL, fatty acid transport protein (FATP) and fatty acid-binding protein (FABP) and protein GPR41, GPR43. Thus, butyrate is involved in body lipid metabolism mainly through the GPR41-mediated ERK-AMPK pathway. Inhibits lipid synthesis in adipose tissue and the liver and promotes lipolysis, avoiding obesity caused by diets with higher fat content. Highlights Butyrate affects lipid metabolism in the body by affecting lipid synthesis/decomposition in adipose tissue and liver. Butyrate mainly through GPR41-mediated ERK-AMPK pathway, inhibit lipid synthesis, promote lipid decomposition, and avoid a large amount of fat accumulation induced by higher-fat diet.

Taxifolin Alleviates DSS-Induced Ulcerative Colitis by Acting on Gut Microbiome to Produce Butyric Acid.

Taxifolin is a bioflavonoid which has been used to treat Inflammatory Bowel Disease. However, taxifolin on DSS-induced colitis and gut health is still unclear. Here, we studied the effect of taxifolin on DSS-induced intestinal mucositis in mice. We measured the degree of intestinal mucosal injury and inflammatory response in DSS treated mice with or without taxifolin administration and studied the changes of fecal metabolites and intestinal microflora using 16S rRNA. The mechanism was further explored by fecal microbiota transplantation. The results showed that the weight loss and diarrhea score of the mice treated with taxifolin decreased in DSS-induced mice and longer colon length was displayed after taxifolin supplementation. Meanwhile, the expression of GPR41 and GPR43 in the colon was significantly increased by taxifolin treatment. Moreover, the expression of TNF-α, IL-1β, and IL-6 in colon tissue was inhibited by taxifolin treatment. The fecal metabolism pattern changed significantly after DSS treatment, which was reversed by taxifolin treatment. Importantly, taxifolin significantly increased the levels of butyric acid and isobutyric acid in the feces of DSS-treated mice. In terms of gut flora, taxifolin reversed the changes of Akkermansia, and further decreased uncultured_bacterium_f_Muribaculaceae. Fecal transplantation from taxifolin-treated mice showed a lower diarrhea score, reduced inflammatory response in the colon, and reduced intestinal mucosal damage, which may be related to the increased level of butyric acid in fecal metabolites. In conclusion, this study provides evidence that taxifolin can ameliorate DSS-induced colitis by altering gut microbiota to increase the production of SCFAs.

Upregulation of G-protein Coupled Receptor 120 in Rats Following Spinal Cord Injury.

G protein-coupled receptors (GPCRs) are fundamental mediators of a wide array of processes including proliferation, immune cell function and neural signaling. GPR120 is a GPCR present within the spleen, lungs, adipose tissue and intestines that is stimulated by endogenous free fatty acids (FFAs). Whether GPR120 is expressed or functionally relevant in the central nervous system (CNS), however, has yet to be directly examined. Herein, a rat spinal cord injury (SCI) model was established and used to explore the expression of GPR120 in SCI. Western blotting and immunohistochemical staining revealed that GPR120 was detectable in the spinal tissues of healthy rats, and the levels rose following SCI and reached the peak on day 3, whereafter they declined to basal levels within two weeks post-SCI. Dual immunofluorescent staining revealed detectable GPR120 expression in astrocytes, microglia and a limited number of neurons. Following SCI, GPR120 upregulation was primarily evident in astrocytes. After injury, colocalization between GPR120 and the proliferative marker PCNA was also detected. Together, these results offer new insights regarding the dynamics of spinal cord GPR120 expression and suggest that it may play important roles in the CNS following SCI.

P088 Metabolomic analysis reveals differences among UC and non-IBD human colonic resections: role of GPCRs

Abstract Background Metabolomics is a recent technique that has bounced into Inflammatory Bowel Diseases (IBD) due to its capacity to elucidate specific metabolites involved in the pathology and changes in the metabolomic profile have been detected in urine, blood or feces from UC patients. G-protein coupled receptors (GPCRs) have been recently identified as promising pharmacological targets. We aim to characterize the metabolomic profile and metabolite-sensing GPCRs expression in colonic resections from UC patients. Methods Colonic resections from UC (n=18) and non-IBD (n=20) patients were obtained. Metabolomic analysis was performed using Nuclear Magnetic Resonance (NMR) to study polar metabolites and Gas-Chromatography or Liquid-Chromatography Mass-Spectrometry to study non-polar metabolites. Results are expressed as μg of metabolite per gram of tissue. Gene expression of GPCRs was analyzed by qPCR. Data were expressed as fold induction vs control (mean±SEM) and compared by a t-test. A p-value&amp;lt;0.05 was considered statistically significant. Pearson correlation matrix were performed with R or mixOmics library in R. Results Metabolomic analysis revealed in colon of UC patients significant increased levels of propionic acid (7.7±1.1), undecanoic acid (0.3±0.02) decanoic acid (4.5±0.9), myristic acid (64.5±8.7), α-linoleic acid (23.1±4.0), aspartic acid (33.9±3.4), phenylalanine (8.8±1.6), glutamic acid, (136.3±10.8), b-hydroxybutyric (8.3±0.9) acid and lactic acid (297.5±34.7) vs non-IBD patients. Gene expression of GPR43 (22.7±9.5), GPR41 (4.7±1.7), GPR109a (19.1±5.3), GPR109b (13.6±4.6), GPR91 (13.2±5.7), GPR84 (6.7±2.0), GPR40 (6.6±1.8), GPR65 (3.0±0.5) and GPR68 (4.6±1.0) were significantly increased in UC patients compared with controls. In contrast, GPR120 (0.9±0.3), GPR119 (0.6±0.2) and GPR35 (0.5±0.1) were significantly reduced in UC patients. N-oleylethanolamide positively correlates with most of GPCRs, especially GPR68, GPR84 and GPR91 while acetic and succinic acids negatively correlate with GPR4 (Fig.1A). In UC patients, medium-chain fatty acids (FA) and most long-chain FA positively correlate with GPR84, GPR142, GPR43, GPR109a, GPR109b, GPR91 and GPR132 (Fig. 1C). Fig1. Graphs show the correlations between data relative to mRNA expression of GPCRs (expressed as ΔCt) vs metabolites. (A) Data from controls and UC. (B) Only controls. (C) Only UC. Conclusion Increased levels of several metabolites and GPCRs are detected in the colon of UC patients and their correlations suggest potential candidates to be involved in UC pathophysiology.

GPR120 prevents colorectal adenocarcinoma progression by sustaining the mucosal barrier integrity

GPR120 (encoded by FFAR4 gene) is a receptor for long chain fatty acids, activated by ω-3 Polyunsaturated Fatty Acids (PUFAs), and expressed in many cell types. Its role in the context of colorectal cancer (CRC) is still puzzling with many controversial evidences. Here, we explored the involvement of epithelial GPR120 in the CRC development. Both in vitro and in vivo experiments were conducted to mimic the conditional deletion of the receptor from gut epithelium. Intestinal permeability and integrity of mucus layer were assessed by using Evans blue dye and immunofluorescence for MUC-2 protein, respectively. Microbiota composition, presence of lipid mediators and short chain fatty acids were analyzed in the stools of conditional GPR120 and wild type (WT) mice. Incidence and grade of tumors were evaluated in all groups of mice before and after colitis-associated cancer. Finally, GPR120 expression was analyzed in 9 human normal tissues, 9 adenomas, and 17 primary adenocarcinomas. Our work for the first time highlights the role of the receptor in the progression of colorectal cancer. We observed that the loss of epithelial GPR120 in the gut results into increased intestinal permeability, microbiota translocation and dysbiosis, which turns into hyperproliferation of epithelial cells, likely through the activation of β -catenin signaling. Therefore, the loss of GPR120 represents an early event of CRC, but avoid its progression as invasive cancer. these results demonstrate that the epithelial GPR120 receptor is essential to maintain the mucosal barrier integrity and to prevent CRC developing. Therefore, our data pave the way to GPR120 as an useful marker for the phenotypic characterization of CRC lesions and as new potential target for CRC prevention.

The expression of nutrient chemosensing gate molecules in the ileum and colon is altered for goats fed on a high-grain diet.

Limited knowledge is clarified about alterations in the related expression of nutrient chemosensors in the distal small intestine and hindgut under a high-grain (HG) diet in small ruminants. Herein, this study was performed to investigate the expression changes related to nutrient sensing and transport in the ileal and colonic epithelium of goats in response to feeding an HG diet. Twelve Liuyang black goats (similar age and weight) were randomly assigned into two groups: an HG diet (concentrate: hay = 90:10) and a CON diet (concentrate: hay = 55:45). Immunohistochemistry was applied to detect morphological changes in the gut epithelium together with altered expression of chemosensors in the ileum and colon. The results showed that feeding an HG diet increased ileal villus height and depth and induced mucosal sloughing in the colon. The expressions of the nutrient transporters GLUT2, GLUT5, SGLT2, CD36, rBAT, EAAT3, and LAT2 and sensing receptors GPR43 and T1R1 were promoted in the ileum under HG conditions. Moreover, feeding an HG diet also enhanced the expression of GLUT2, SGLT2, CD36, and GPR43 in the colon. These findings indicate that adaptation of the gastrointestinal tract to the HG diet promoted the absorption of glucose, fatty acids, and amino acids in goats.

GPR84 signaling promotes intestinal mucosal inflammation via enhancing NLRP3 inflammasome activation in macrophages

The putative medium-chain free fatty acid receptor GPR84 is a G protein-coupled receptor primarily expressed in myeloid cells that constitute the innate immune system, including neutrophils, monocytes, and macrophages in the periphery and microglia in the brain. The fact that GPR84 expression in leukocytes is remarkably increased under acute inflammatory stimuli such as lipopolysaccharide (LPS) and TNFα suggests that it may play a role in the development of inflammatory and fibrotic diseases. Here we demonstrate that GPR84 is highly upregulated in inflamed colon tissues of active ulcerative colitis (UC) patients and dextran sulfate sodium (DSS)-induced colitis mice. Infiltrating GPR84+ macrophages are significantly increased in the colonic mucosa of both the UC patients and the mice with colitis. Consistently, GPR84−/− mice are resistant to the development of colitis induced by DSS. GPR84 activation imposes pro-inflammatory properties in colonic macrophages through enhancing NLRP3 inflammasome activation, while the loss of GPR84 prevents the M1 polarization and properties of proinflammatory macrophages. CLH536, a novel GPR84 antagonist discovered by us, suppresses colitis by reducing the polarization and function of pro-inflammatory macrophages. These results define a unique role of GPR84 in innate immune cells and intestinal inflammation, and suggest that GPR84 may serve as a potential drug target for the treatment of UC.

Kcnk3, Ggta1, and Gpr84 are involved in hyperbaric oxygenation preconditioning protection on cerebral ischemia\u2013reperfusion injury

The present study aimed to explore the potential mechanism of the effect of hyperbaric oxygenation (HBO) preconditioning on cerebral ischemia and reperfusion injury (CIRI). GSE23160 dataset was used to identify differentially expressed genes (DEGs) from striatum between the middle cerebral artery occlusion (MCAO)/reperfusion and sham rats. The gene clusters with continuous increase and decrease were identified by soft clustering analysis in Mfuzz, and functional enrichment analysis of these genes was performed using clusterProfiler package. The intersection set of the genes with significantly altered expression at post-reperfusion 2, 8, and 24 h were screened in comparison to 0 h (sham group), and the expression of these genes was detected in the MCAO/reperfusion model and HBO preconditioning groups by real-time PCR (RT-PCR) and western blotting. A total of 41 upregulated DEGs, and 7 downregulated DEGs were detected, among which the expression of Gpr84 and Ggta1 was significantly upregulated at each reperfusion phase as compared to the sham group, while the expression of Kcnk3 was significantly downregulated except in the postreperfusion 8 h in the striatum group. RT-PCR and western blotting analyses showed that the expression of Ggta1, Gpr84, and Kcnk3 genes between the MCAO/reperfusion and sham rats were consistent with the bioinformatics analysis. In addition, the HBO preconditioning reduced the expression of Ggta1 and Gpr84 and increased the expression of Kcnk3 in MCAO/reperfusion rats. Kcnk3, Ggta1, and Gpr84 may play a major role in HBO-mediated protection of the brain against CIRI.

GPR120 Inhibits Colitis Through Regulation of CD4+ T Cell Interleukin 10 Production

G protein-coupled receptor (GPR) 120 has been implicated in regulating metabolic syndromes with anti-inflammatory function. However, the role of GPR120 in intestinal inflammation is unknown. Here, we investigated whether and how GPR120 regulates CD4+ T cell function to inhibit colitis development. Dextran sodium sulfate (DSS)-induced colitis model, Citrobacter rodentium infection model, and CD4+ T cell adoptive transfer model were used to analyze the role of GPR120 in regulating colitis development. The effect of GPR120 on CD4+ T cell functions was analyzed by RNA sequencing, flow cytometry, and Seahorse metabolic assays. Mice were administered GPR120 agonist for investigating the potential of GPR120 agonist in preventing and treating colitis. Deficiency of GPR120 in CD4+ T cells resulted in more severe colitis in mice upon dextran sodium sulfate insult and enteric infection. Transfer of GPR120-deficient CD4+CD45Rbhi T cells induced more severe colitis in Rag-/- mice with lower intestinal interleukin (IL) 10+CD4+ Tcells. Treatment with the GPR120 agonist CpdA promoted CD4+ T cell production of IL10 by up-regulating Blimp1 and enhancing glycolysis, which was regulated by mTOR. GPR120 agonist-treated wild-type, but not IL10-deficient and Blimp1-deficient, T helper 1 cells induced less severe colitis. Furthermore, oral administration of GPR120 agonist protected mice from intestinal inflammation in both prevention and treatment schemes. Gpr120 expression was positively correlated with Il10 expression in the human colonic mucosa, including patients with inflammatory bowel diseases. Our findings show the role of GPR120 in regulating intestinal CD4+ T cell production of IL10 to inhibit colitis development, which identifies GPR120 as a potential therapeutic target for treating inflammatory bowel diseases.

The Gut Microbiota and Their Metabolites in Human Arterial Stiffness

Gut microbiota-derived metabolites, such as short-chain fatty acids (SCFAs) have vasodilator properties in animal and human exvivo arteries. However, the role of the gut microbiota and SCFAs in arterial stiffness in humans is still unclear. Here we aimed to determine associations between the gut microbiome, SCFA and their G-protein coupled sensing receptors (GPCRs) in relation to human arterial stiffness. Ambulatory arterial stiffness index (AASI) was determined from ambulatory blood pressure (BP) monitoring in 69 participants from regional and metropolitan regions in Australia (55.1% women; mean, 59.8± SD, 7.26 years of age). The gut microbiome was determined by 16S rRNA sequencing, SCFA levels by gas chromatography, and GPCR expression in circulating immune cells by real-time PCR. There was no association between metrics of bacterial α and β diversity and AASI or AASI quartiles in men and women. We identified two main bacteria taxa that were associated with AASI quartiles: Lactobacillus spp. was only present in the lowest quartile, while Clostridium spp. was present in all quartiles but the lowest. AASI was positively associated with higher levels of plasma, but not faecal, butyrate. Finally, we identified that the expression of GPR43 (FFAR2) and GPR41 (FFAR3) in circulating immune cells were negatively associated with AASI. Our results suggest that arterial stiffness is associated with lower levels of the metabolite-sensing receptors GPR41/GPR43 in humans, blunting its response to BP-lowering metabolites such as butyrate. The role of Lactobacillus spp. and Clostridium spp., as well as butyrate-sensing receptors GPR41/GPR43, in human arterial stiffness needs to be determined.

The effects of increasing fruit and vegetable intake in children with asthma: A randomized controlled trial.

A high fruit and vegetable (F&V) diet reduces asthma exacerbations in adults; this has not been examined in children to date. To investigate the effect of a 6-month, high F&V diet on the time to first asthma exacerbation in children with asthma, in a parallel-group, randomized, controlled trial. Children (aged 3-11years) with asthma, history of exacerbations and usual low F&V intake (≤3 serves/day) were randomized to the intervention (high F&V diet) or control group (usual diet) for 6months. The primary outcome was time to first exacerbation requiring medical intervention. Secondary outcomes included exacerbation rate, lung function, plasma TNF-α, CRP, and IL-6, faecal microbiota and peripheral blood mononuclear cell (PBMC) histone deacetylase (HDAC) activity and G-protein coupled receptor (GPR) 41/43 and HDAC (1-11) expression. 67 children were randomized between September 2015 and July 2018. F&V intake (difference in change (∆): 3.5serves/day, 95% CI: [2.6, 4.4] p<0.001) and plasma total carotenoids (∆: 0.44µg/ml [0.19, 0.70] p=0.001) increased after 6months (intervention vs control). Time to first exacerbation (HR: 0.81, 95% CI: [0.38, 1.69], p=0.569; control vs. intervention) and exacerbation rate (IRR: 0.84, [0.47, 1.49], p=0.553; control vs. intervention) were similar between groups. In per-protocol analysis, airway reactance z-scores increased in the intervention versus control group (X5 ∆: 0.76 [0.04, 1.48] p=0.038, X20 ∆: 0.93 [0.23, 1.64] p=0.009) and changes in faecal microbiota were observed though there was no difference between groups in systemic inflammation or molecular mechanisms. In the control group, CRP and HDAC enzyme activity increased, while GPR41 expression decreased. No adverse events attributable to the interventions were observed. A high F&V diet did not affect asthma exacerbations over the 6-month intervention, though warrants further investigation as a strategy for improving lung function and protecting against systemic inflammation in children with asthma.

Synthesis, molecular docking, dynamic simulation and pharmacological characterization of potent multifunctional agent (dual GPR40-PPARγ agonist) for the treatment of experimental type 2 diabetes

The current manuscript describes two molecules that were designed against PPARγ and GPR40 receptors. The preparation of the compounds was carried out following a synthetic route of multiple steps. Then, the mRNA expression levels of PPARγ, GLUT4, and GPR40 induced by compounds were measured and quantified in adipocyte and β-pancreatic cell cultures. The synthesized compound 1 caused an increase in the 4-fold expression of mRNA of PPARγ regarding the control and had a similar behavior to the pioglitazone, while compound 2 only increased 2-fold the expression. Also, the compound 1 increased to 7-fold the GLUT4 expression levels, respect to the control and twice against the pioglitazone. On the other hand, the 1 increase 3-fold GPR40 expression, and compound 2 had a minor activity. Besides, 1 and 2 showed a moderated increase on insulin secretion and calcium mobilization versus the glibenclamide. Based on the molecular docking studies, the first compound had a similar conformation to co-crystal ligands into the binding site of both receptors. The poses were docked keeping the most important interactions and maintaining the interaction along the Molecular Dynamics simulation (20 ns). Finally, compound (1) showed an antihyperglycemic effect at 5 mg/kg, however at higher doses of 25 mg/kg it controlled blood glucose levels associated with feeding intake and without showing the adverse effects associated with insulin secretagogues (hypoglycemia). For these reasons, we have concluded that molecule 1 acts as a dual PPARγ and GPR40 agonist offering a better glycemic control than current treatments.

Exposure to ethephon compromises endometrial decidualization in mice during early pregnancy via GPR120

Exposure to ethephon (ETH), a plant growth regulator commonly used for several purposes, can potentially decrease sperm numbers and viability. Occasional findings regarding ETH effects on female reproduction during early pregnancy have also been reported. During early pregnancy, endometrial decidualization is a critical event for embryo implantation and pregnancy maintenance. Thus, we aimed to explore the effect and mechanism of ETH on endometrial decidualization both in vivo and in vitro. Mice were gavaged with 0 and 285 mg/kg b.w. ETH from gestational days (GD)1 until sacrifice, whereas pseudopregnant mice from pseudopregnant day 1 (PPD-1) until PPD-8. Primary mouse endometrial stromal cells (mESCs) received 640 ug/ml ETH and added E2 and P4 to induce decidualization. Results indicated female albino CD1 mice exposed to high dose of ETH (285 mg/kg b.w.) by oral gavage, the number of embryo implantation sites on GD6 and GD8 were significantly decreased, the levels of serum E2 and P4 on GD8 were significantly decreased. Compared with the control group, the decidualization response artificially induced by corn oil in pseudopregnant mice and by E2 and P4 in primary mouse endometrial stromal cells (mESCs) was weakened in the high dose of ETH treated group. The high dose, 285 mg/kg b.w ETH treated group altered the expression of endometrial decidual markers on GD6 and GD8. The triglyceride and fatty acid metabolism-related genes were significantly increased after female albino CD1 mice exposed to high does, 285 mg/kg b.w ETH on GD6 and GD8. GPR120 was substantially reduced after ETH treatment. When overexpression of GPR120, the compromised decidualization induced by ETH treatment was rescued. Furthermore, molecular docking presented Thr234 and His251 of GPR120 as preferred binding sites for ETH. Mutation of these two sites rescued the compromised decidualization induced by ETH. In conclusion, we demonstrated that ETH exposure could impair decidualization during early pregnancy. GPR120 expression and binding between GPR120 and ETH are crucial for impaired decidualization mediated via ETH.

Lipotoxicity induced pancreatic beta cell damage is associated with GPR119/MST1/FoxO1 Pathway

IntroductionG-protein-coupled receptor 119 (GPR119) is emerging as a potential therapeutic target against type 2 diabetes with beneficial effects on glucose homeostasis. However, the function of GRP119 in lipotoxicity induced pancreatic beta cell apoptosis and the molecular mechanism remains largely unknown.Material and methodsImpact of GPR119 on pancreatic islet beta cell apoptosis was evaluated in INS-1 cells treated with palmitate. The subsequent modulation of the MST1-FOXO1-Pdx1 signaling pathway and pro-apoptotic caspase-3 system were determined by measuring the target protein and mRNA expression. Dyslipidemia mice with gain and loss of GPR119 function by the application of specific lenti-viral vector was utilized to evaluate the impact of GPR119 on pancreas function in vivo. Lipid metabolism, glucose and insulin response, morphological changes as well as activation/inhibition of MST1-FOXO1-Pdx1 signaling pathway in pancreas were analyzed systematically.ResultsPalmitate treatment stimulated pro-apoptotic response in INS-1 cells, accompanied by inhibition of GPR119 expression and the subsequent activation of the MST1-FOXO1 combined with inhibition of Pdx1 signaling cascade. Activation of GPR119 by MBX prevents INS-1 cell from lipotoxicity induced apoptosis by targeting the MST1-FOXO1-Pdx1 pathway. Moreover, overexpression of GPR119 significantly attenuates the dyslipidemia and dysfunction of the pancreas. In contrast, inactivation of GPR119 by lentiviral vector in mice results in accelerated pancreas apoptosis and malfunction. The protective effects of GRP119 on lipotoxicity induced pancreas dysfunction are associated with modulating the MST1-FOXO1-Pdx1 signaling cascade.ConclusionsGPR119 alleviates lipotoxicity induced pancreatic beta cell apoptosis and malfunction through regulating MST1-FOXO1-Pdx1 signaling pathway.