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Tu1840 Protective Role of Uric Acid Excretion to the Intestinal Tract on Small Intestinal Injury Induced by Indomethacin
Tu1838
Table 1 The baseline charecteristics of study population
AGA Abstracts
Gene Expression in Luminal Extracellular Vesicles (EVS) in Patients With Inflammatory Bowel Disease (IBD) Shuji Mitsuhashi, Masato Mitsuhashi, Amanda Ting, Adam S. Cheifetz, Harry Anastopoulos, Simon C. Robson, Alan C. Moss Background: Extracellular Vesicles (EVs) are nanoparticles released from many cells as intercellular signaling vehicles. They have not previously been described in IBD. We sought to characterize the nature and function of EVs obtained from the intestinal lumen of patients with IBD. Methods: Intestinal luminal fluid (ILF) was aspirated (10mL) during routine endoscopic procedures from healthy controls (HC) and patients with IBD. EVs were isolated from ILF using an EV collection filter tube. EV size and concentration from representative samples were determined using nanoparticle tracking. Gene expression in EVs was quantified using RT-PCR of RNA isolated from EVs. Labeled RNA was used to track the uptake of EVs by intestinal epithelial cells. Results: Samples from healthy controls (n=11), patients with Crohn's Disease (CD, n=13) and Ulcerative Colitis (UC, n=21) were analyzed. Isolated EVs had mean size of 150nm, and a concentration of 5 x 1011 particles/mL. There were significant differences in mRNA levels of epithelial defense genes (EPCAM, MUC2, TFF1, DEFA3) and leukocyte activation markers (CD45, TGFB1, S100A9) between EVs from HCs and IBD patients (p<0.05 by t-test). Significant differences in gene expression between samples from patients with higher endoscopic severity score (Mayo ≥2) were also noted. Co-culture of intestinal epithelial cell lines with EVs from HCs and patients with IBD led to uptake by epithelial cells of these EVs. In response, there was a significant up-regulation of IL-8 mRNA from cells cultured with EVs from IBD patients, when compared to cells cultured with HC EVs. Conclusions: These data identify abundant EVs in the lumen of patients with IBD for the first time. These EVs exhibit significant differences in mRNA content according to disease activity. We also demonstrate an impact on epithelial cells from absorbed luminal EVs. These findings warrant further investigation on the immunomodulatory properties of luminal EVs. Tu1839 Pulse Wave Velocity, Intima Media Thickness and Flow Mediated Dilatation in Patients With Normotensive Normoglysemic Inflammatory Bowel Disease Kadir Ozturk, Ahmet K. Guler, Mehmet Cakir, Hakan Demirci, Ahmet Uygun, Mustafa Gulsen, Sait Bagci Introduction: IBD patients have lower traditional cardiovascular risk factors than general population. However, some studies suggested that IBD patients have increased risk for cardiovascular events and atherosclerosis. The cause of increased cardiovascular risk is not fully understood. Chronic inflammatory process may predispose to atherosclerosis.In fact, the effect of chronic inflammation on devolepment of atherosclerosis has already been demonstrated in various chronic inflammatory diseases especially rheumatoid arthritis and systemic lupus erythematosus. Aim: We aimed primarly to investigate subclinical atherosclerosis in patients with IBD, by measuring cf-PWV, CIMT and FMD compared to matched normal controls. Our secondary aim was to demonstrate the relationship between these parameters and disease variables (disease duration and extent). Methods: A total of 192 cases, consisting of 74 patients with UC, 52 patients with CD and 66 healthy control subjects, were included in the study. Patients with previous cardiovascular disease, DM, HT, chronic renal failure, infectious and inflammatory disorders other than CD and UC were excluded.High sensitive CRP was measured by using a nephelometric method. Hemoglobin, platelet, white blood cell, serum levels of FPG, uric acid, high density lipoprotein, triglycerides and creatinine were determined by using auto-analyzers. LDL was calculated by using the Friedewald's formula. PWV was calculated using an automatic device. Measurements of FMD and CIMT were performed with B-mode ultrasonography, by using a high resolution, 18 MHz linear-array transducer. Results: The baseline charecteristics of study population are summarized in Table 1. The cf-PWV levels were 8.13±1.61 m/s in the UC patients, 8.16±1.74 m/s in the CD patients and 6.85±0.95 m/s in the healthy subjects. The levels of cf-PWV were significantly higher in CD and UC patients than control groups (P<0.001), but there was no significantly difference between UC and CD (P>0.05). We found significantly decreased FMD levels in patients with UC and CD (9.6±5.1% vs 10.8±4.4%), when compared with control subjects (15.1±9.7%) (P<0.05), but FMD levels did not differ significantly between UC and CD patients (p>0.05). On the other hand, no difference in CIMT was detected between UC, CD and controls groups. We did not find any significant relationships between disease extent, cf-PWV, FMD and CIMT (P>0.05). There was a significant correlation between cf-PWV and disease duration (P=0.001, r=0.297). Conclusion: The present study suggest that IBD patients, without traditional cardiovascular risk factors have increased risk of endothelial dysfunction and atherosclerosis. Structural changes in arterial vessel wall occur because of long-term exposure to inflammation or cardiovascular risk factors after endothelial dysfunction.
AGA Abstracts
UC ulcerative colitis, CD crohn disease, BMI body mass index, SBP systolic blood pressure, DBP diastolic blood pressure, hs-CRP High sensitive c reactive protein, ESR erytrocyte sedimentation rate, HGB hemoglobine, PLT platelets, WBC White blood cells, FPG fasting plasma glucose, LDL C, low density lipoprotein cholesterol, HDL C high density lipoprotein cholesterol, TG triglyceride, anti-TNF anti tumor necrosis factor, cf-PWV carotid femoral pulse wave velocity, CIMT carotid intima-media thickness, FMD flow mediated dilatation Tu1840 Protective Role of Uric Acid Excretion to the Intestinal Tract on Small Intestinal Injury Induced by Indomethacin Yuichi Yasutake, Ryota Hokari, Yasushi Inoue, Takeshi Takajo, Koji Maruta, Hirokazu Sato, Kazuyuki Narimatsu, Chie Kurihara, Yoshikiyo Okada, Kenichi Yoshikawa, Chikako Watanabe, Shunsuke Komoto, Kengo Tomita, Shigeaki Nagao, Soichiro Miura Background: Genome wide association study shows relationship between ATP-binding cassette sub-family G member 2 (ABCG2) and hyperuricemia. Surprisingly, it has been reported that small intestine is another main organ to excrete uric acid (UA) outside the body through ABCG2 in addition to kidney and dysfunction of ABCG2 by SNPs is responsible for hyperuricemia and gout in over 60% of the patients in Japan. Although intestine is directly exposed to a large amount of UA, its role in small intestinal diseases has not been studied yet. It is already known UA plays anti-inflammatory roles as the main antioxidants. Actually, hyperuricemia shows inverse correlation to progress of the neurological diseases such as multiple sclerosis or Parkinson's disease. We investigated whether UA plays antiinflammatory roles in the NSAIDs-induced intestinal injury, which is the most common small intestinal disorders but its treatment has not been well established. Methods: Because mice have uricase to catalyze UA, we treated mice with potassium oxonate (KOX), a uricase inhibitor. To investigate direct exposure to UA on small intestine, male C57BL6 of 8 week old mice were treated with 250mg/kg UA and 500mg/kg KOX by oral gavage and induced ileitis with 10mg/kg indomethacin (IND) i.p. To assess the effect of UA excreted to intestinal lumen through ABCG2 transporter, we made hyperuricemia mice by intraperitoneal administration of 250mg/kg inosinic acid (IMP), UA precursor, and 500mg/kg KOX. Ten mg/kg febxostat (FEB), a xanthine oxidase inhibitor, was administered to clarify the inhibitory effect of IMP catalyzation to UA. Intestinal damage was evaluated by ulcer area after injection of evans blue dye. Inflammatory cytokine was evaluated by mRNA expression in the intestinal mucosa by using real-time RT-PCR. Results: IND treatment induced small intestinal injury with increased expression of mucosal TNF-alpha. Direct exposure to UA on intestine ameliorated intestinal injury (ulcer area of control group: 30.3±5.6 mm2 vs UA treatment: 17.7±2.1 mm2). Intraperitoneal administration of IMP induced hyperuricemia at the concentration of 2.8±0.17 mg/dl. This treatment ameliorated intestinal injury (ulcer area of control group: 34.2±8.1mm2 vs IMP treatment 15.8±3.3 mm2) with decreased mucosal TNF alpha expression. FEB treatment decreased blood concentration of UA and canceled the ameliorating effect of IMP treatment to intestinal injury (ulcer area of control group: 31.6±4.6 mm2, IMP treatment: 13.2±3.6 mm2, FEB treatment alone: 31.8±11.8 mm2, FEB and IMP treatment: 25.8±7.0 mm2). Conclusion: This is the first report showing that UA has a potential modulator of intestinal inflammation induced by IND. We conclude UA plays anti-inflammatory roles in the intestinal tract and ABCG2 may become a possible target for small intestinal inflammation.
S-916
Table 1 The baseline charecteristics of study population
AGA Abstracts
Gene Expression in Luminal Extracellular Vesicles (EVS) in Patients With Inflammatory Bowel Disease (IBD) Shuji Mitsuhashi, Masato Mitsuhashi, Amanda Ting, Adam S. Cheifetz, Harry Anastopoulos, Simon C. Robson, Alan C. Moss Background: Extracellular Vesicles (EVs) are nanoparticles released from many cells as intercellular signaling vehicles. They have not previously been described in IBD. We sought to characterize the nature and function of EVs obtained from the intestinal lumen of patients with IBD. Methods: Intestinal luminal fluid (ILF) was aspirated (10mL) during routine endoscopic procedures from healthy controls (HC) and patients with IBD. EVs were isolated from ILF using an EV collection filter tube. EV size and concentration from representative samples were determined using nanoparticle tracking. Gene expression in EVs was quantified using RT-PCR of RNA isolated from EVs. Labeled RNA was used to track the uptake of EVs by intestinal epithelial cells. Results: Samples from healthy controls (n=11), patients with Crohn's Disease (CD, n=13) and Ulcerative Colitis (UC, n=21) were analyzed. Isolated EVs had mean size of 150nm, and a concentration of 5 x 1011 particles/mL. There were significant differences in mRNA levels of epithelial defense genes (EPCAM, MUC2, TFF1, DEFA3) and leukocyte activation markers (CD45, TGFB1, S100A9) between EVs from HCs and IBD patients (p<0.05 by t-test). Significant differences in gene expression between samples from patients with higher endoscopic severity score (Mayo ≥2) were also noted. Co-culture of intestinal epithelial cell lines with EVs from HCs and patients with IBD led to uptake by epithelial cells of these EVs. In response, there was a significant up-regulation of IL-8 mRNA from cells cultured with EVs from IBD patients, when compared to cells cultured with HC EVs. Conclusions: These data identify abundant EVs in the lumen of patients with IBD for the first time. These EVs exhibit significant differences in mRNA content according to disease activity. We also demonstrate an impact on epithelial cells from absorbed luminal EVs. These findings warrant further investigation on the immunomodulatory properties of luminal EVs. Tu1839 Pulse Wave Velocity, Intima Media Thickness and Flow Mediated Dilatation in Patients With Normotensive Normoglysemic Inflammatory Bowel Disease Kadir Ozturk, Ahmet K. Guler, Mehmet Cakir, Hakan Demirci, Ahmet Uygun, Mustafa Gulsen, Sait Bagci Introduction: IBD patients have lower traditional cardiovascular risk factors than general population. However, some studies suggested that IBD patients have increased risk for cardiovascular events and atherosclerosis. The cause of increased cardiovascular risk is not fully understood. Chronic inflammatory process may predispose to atherosclerosis.In fact, the effect of chronic inflammation on devolepment of atherosclerosis has already been demonstrated in various chronic inflammatory diseases especially rheumatoid arthritis and systemic lupus erythematosus. Aim: We aimed primarly to investigate subclinical atherosclerosis in patients with IBD, by measuring cf-PWV, CIMT and FMD compared to matched normal controls. Our secondary aim was to demonstrate the relationship between these parameters and disease variables (disease duration and extent). Methods: A total of 192 cases, consisting of 74 patients with UC, 52 patients with CD and 66 healthy control subjects, were included in the study. Patients with previous cardiovascular disease, DM, HT, chronic renal failure, infectious and inflammatory disorders other than CD and UC were excluded.High sensitive CRP was measured by using a nephelometric method. Hemoglobin, platelet, white blood cell, serum levels of FPG, uric acid, high density lipoprotein, triglycerides and creatinine were determined by using auto-analyzers. LDL was calculated by using the Friedewald's formula. PWV was calculated using an automatic device. Measurements of FMD and CIMT were performed with B-mode ultrasonography, by using a high resolution, 18 MHz linear-array transducer. Results: The baseline charecteristics of study population are summarized in Table 1. The cf-PWV levels were 8.13±1.61 m/s in the UC patients, 8.16±1.74 m/s in the CD patients and 6.85±0.95 m/s in the healthy subjects. The levels of cf-PWV were significantly higher in CD and UC patients than control groups (P<0.001), but there was no significantly difference between UC and CD (P>0.05). We found significantly decreased FMD levels in patients with UC and CD (9.6±5.1% vs 10.8±4.4%), when compared with control subjects (15.1±9.7%) (P<0.05), but FMD levels did not differ significantly between UC and CD patients (p>0.05). On the other hand, no difference in CIMT was detected between UC, CD and controls groups. We did not find any significant relationships between disease extent, cf-PWV, FMD and CIMT (P>0.05). There was a significant correlation between cf-PWV and disease duration (P=0.001, r=0.297). Conclusion: The present study suggest that IBD patients, without traditional cardiovascular risk factors have increased risk of endothelial dysfunction and atherosclerosis. Structural changes in arterial vessel wall occur because of long-term exposure to inflammation or cardiovascular risk factors after endothelial dysfunction.
AGA Abstracts
UC ulcerative colitis, CD crohn disease, BMI body mass index, SBP systolic blood pressure, DBP diastolic blood pressure, hs-CRP High sensitive c reactive protein, ESR erytrocyte sedimentation rate, HGB hemoglobine, PLT platelets, WBC White blood cells, FPG fasting plasma glucose, LDL C, low density lipoprotein cholesterol, HDL C high density lipoprotein cholesterol, TG triglyceride, anti-TNF anti tumor necrosis factor, cf-PWV carotid femoral pulse wave velocity, CIMT carotid intima-media thickness, FMD flow mediated dilatation Tu1840 Protective Role of Uric Acid Excretion to the Intestinal Tract on Small Intestinal Injury Induced by Indomethacin Yuichi Yasutake, Ryota Hokari, Yasushi Inoue, Takeshi Takajo, Koji Maruta, Hirokazu Sato, Kazuyuki Narimatsu, Chie Kurihara, Yoshikiyo Okada, Kenichi Yoshikawa, Chikako Watanabe, Shunsuke Komoto, Kengo Tomita, Shigeaki Nagao, Soichiro Miura Background: Genome wide association study shows relationship between ATP-binding cassette sub-family G member 2 (ABCG2) and hyperuricemia. Surprisingly, it has been reported that small intestine is another main organ to excrete uric acid (UA) outside the body through ABCG2 in addition to kidney and dysfunction of ABCG2 by SNPs is responsible for hyperuricemia and gout in over 60% of the patients in Japan. Although intestine is directly exposed to a large amount of UA, its role in small intestinal diseases has not been studied yet. It is already known UA plays anti-inflammatory roles as the main antioxidants. Actually, hyperuricemia shows inverse correlation to progress of the neurological diseases such as multiple sclerosis or Parkinson's disease. We investigated whether UA plays antiinflammatory roles in the NSAIDs-induced intestinal injury, which is the most common small intestinal disorders but its treatment has not been well established. Methods: Because mice have uricase to catalyze UA, we treated mice with potassium oxonate (KOX), a uricase inhibitor. To investigate direct exposure to UA on small intestine, male C57BL6 of 8 week old mice were treated with 250mg/kg UA and 500mg/kg KOX by oral gavage and induced ileitis with 10mg/kg indomethacin (IND) i.p. To assess the effect of UA excreted to intestinal lumen through ABCG2 transporter, we made hyperuricemia mice by intraperitoneal administration of 250mg/kg inosinic acid (IMP), UA precursor, and 500mg/kg KOX. Ten mg/kg febxostat (FEB), a xanthine oxidase inhibitor, was administered to clarify the inhibitory effect of IMP catalyzation to UA. Intestinal damage was evaluated by ulcer area after injection of evans blue dye. Inflammatory cytokine was evaluated by mRNA expression in the intestinal mucosa by using real-time RT-PCR. Results: IND treatment induced small intestinal injury with increased expression of mucosal TNF-alpha. Direct exposure to UA on intestine ameliorated intestinal injury (ulcer area of control group: 30.3±5.6 mm2 vs UA treatment: 17.7±2.1 mm2). Intraperitoneal administration of IMP induced hyperuricemia at the concentration of 2.8±0.17 mg/dl. This treatment ameliorated intestinal injury (ulcer area of control group: 34.2±8.1mm2 vs IMP treatment 15.8±3.3 mm2) with decreased mucosal TNF alpha expression. FEB treatment decreased blood concentration of UA and canceled the ameliorating effect of IMP treatment to intestinal injury (ulcer area of control group: 31.6±4.6 mm2, IMP treatment: 13.2±3.6 mm2, FEB treatment alone: 31.8±11.8 mm2, FEB and IMP treatment: 25.8±7.0 mm2). Conclusion: This is the first report showing that UA has a potential modulator of intestinal inflammation induced by IND. We conclude UA plays anti-inflammatory roles in the intestinal tract and ABCG2 may become a possible target for small intestinal inflammation.
S-916