- Email: [email protected]
P240 BILBERRY EXTRACT ATTENUATES DEVELOPMENT OF NONALCOHOLIC STEATOHEPATITIS IN ApoE3L MICE
POSTERS Results: Hepatic steatosis was not altered in CD36 KO mice fed a HFD compared to WT mice. However, de novo lipogenesis (DNL) was reduced in CD36 KO mice. In line with this, SCD1 desaturase activity were significantly reduced in CD36 KO mice compared to WT littermates. In addition, the levels of the anti-inflammatory polyunsaturated omega-3 fatty acids, AlphaLinolenic Acid (C18:3W3) and eicosapentaenoic acid (C20:5W3) were reduced, and the pro-inflammatory polyunsaturated omega-6 fatty acid Docosapentaenate (C22:5W6) was increased, suggesting that CD36 ablation contributes to a hepatic inflammation. This was confirmed by pathological assessment of liver sections and RT-PCR analysis of several inflammatory cytokines and genes. Conclusions: Our data indicates that CD36 ablation is associated with increased hepatic inflammation that may be mediated by a reduction in anti-inflammatory lipid species present. Therefore, CD36 may serve as a protective mechanism in the liver, dampening liver inflammation and attenuating the progression towards NASH. P238 METABOLICALLY INDUCED LIVER INFLAMMATION PROMOTES NASH DEVELOPMENT AND DIFFERS FROM ENDOTOXIN- AND CYTOKINE-INDUCED INFLAMMATION W. Liang1 , J.H. Lindeman2 , A. Menke3 , D. Koonen4 , P.Y. Wielinga1 , M. Morrison1 , P. Mulder1 , L.A.M. Havekes5 , A.M. van den Hoek1 , R. Kleemann1 . 1 Metabolic Health Research, The Dutch Organization for Applied Scientific Research (TNO), 2 Department of Vascular Surgery, Leiden University Medical Center, Leiden, 3 TNO-Triskelion, Zeist, 4 Molecular Genetics, University Medical Center Groningen, Groningen, 5 Departments of Endocrinology and Cardiology, Leiden University Medical Center, Leiden, Netherlands E-mail: [email protected] Background and Aims: The exact nature of the chronic inflammatory component that drives the progression of bland liver steatosis (BS) to non-alcoholic hepatosteatitis (NASH) is unclear. Among the possible triggers of inflammation are endotoxins, inflammatory cytokines as well as diet-related metabolic inducers of inflammation. In this study, we investigated the effect of LPS, IL1b (via minipumps), carbohydrate and cholesterol (via diet) on the progression from BS to NASH using human-ApoE3L.CETP transgenic mice. Hallmarks of human NASH (neutrophil infiltration, NF-úBactivation) were defined in biopsy material to guide subsequent histopathological analyses in mice. Methods: Experimental BS was established after 10 weeks of high fat diet (HFD) feeding. HFD was continued and inflammatory triggers (LPS, IL1b, carbohydrate or cholesterol) were superimposed for six more weeks, or not (control). Results: Livers of HFD-fed mice did not develop NASH and remained at the level of BS. All four inflammatory triggers activated intrahepatic NF-úB significantly and comparably (≥5-fold). Despite this, HFD+LPS and HFD+IL1b did not induce a NASHlike phenotype. By contrast, mice treated with HFD+carbohydrate or HFD+cholesterol developed human-like NASH characterized by greatly increased steatosis, hepatocellular hypertrophy and formation of mixed-type inflammatory foci containing neutrophils. Infiltration of neutrophils and an activation of the pro-inflammatory transcription factor AP-1 in liver was a specific and common effect of the NASH-inducing triggers and not observed with LPS or IL1b. Conclusions: HFD-feeding followed by LPS or IL1b-induced NF-úB activation per se is not sufficient to promote a transition from BS to NASH. HFD-feeding followed by metabolically-evoked inflammation induces additional inflammatory components which promote NASH.
P239 METABOLIC SYNDROME AND NON-ALCOHOLIC STEATOHEPATITIS (NASH) ARE ATTENUATED BY INTERVENTION WITH CASPASE-1 INHIBITOR IN HIGH FAT DIET FED LDLR−/− .LEIDEN MICE P.Y. Wielinga, K. Salic, W. Liang, M. Morrison, P. Mulder, T. Kooistra, R. Kleemann. Metabolic Health Research, The Dutch Organization for Applied Scientific Research (TNO), Leiden, Netherlands E-mail: [email protected] Background and Aims: Non-alcoholic steatohepatitis (NASH) is a serious liver pathology which develops as a complication of the metabolic syndrome. Recently, the inflammasome was proposed to be involved in the development of NASH. Here we investigate whether a chemical inhibitor of caspase-1 in already manifest metabolic syndrome would prevent the development to NASH. Methods: Male LDLR−/− .Leiden mice were fed a high fat diet (HFD; group 1) or low fat diet (LFD; group 2) for 21 weeks. In a third group, intervention with caspase-1 inhibitor Ac-YVADCMK (40 mg/kg daily) was started after 9 weeks of HFD (manifest metabolic syndrome) and continued until 21 weeks. Results: HFD treated mice developed obesity and insulin resistance after already 9 weeks of HFD feeding. Intervention with caspase-1 inhibitor attenuated a further development of insulin resistance, and reduced body weight gain as well as adipose tissue inflammation compared to HFD. Histopathological analysis of the livers clearly demonstrated prevention of NASH development with caspase-1 inhibitor, livers were less steatotic and neutrophil infiltration was diminished. Additionally, hepatic fibrosis quantified by sirius red staining and acta2 and col1a1 gene expression as observed in HFD treated mice, was completely prevented. Conclusions: Intervention with a caspase-1 inhibitor in already established disease improved hallmarks of the metabolic syndrome and prevented the development of NASH. Our data further support the importance of caspase-1/inflammasome in the development of NASH and demonstrate that therapeutic intervention in the already ongoing disease process is feasible. P240 BILBERRY EXTRACT ATTENUATES DEVELOPMENT OF NONALCOHOLIC STEATOHEPATITIS IN ApoE3L MICE M.C. Morrison1,2,3 , W. Liang1 , K. Salic1 , P. Mulder1 , P. Wielinga1 , T. Kooistra1 , P. Heeringa2 , R. Kleemann1 . 1 Department of Metabolic Health Research, Netherlands Organization for Applied Scientific Research (TNO), Leiden, 2 Department of Pathology and Medical Biology, University Medical Center Groningen, Groningen, 3 Top Institute Food and Nutrition, Wageningen, Netherlands E-mail: [email protected] Background and Aims: Anthocyanin-rich extract of bilberry fruits may have beneficial effects on lipid metabolism and inflammation and was demonstrated to have hepatoprotective properties in models of restraint-stress- and chemically-induced liver damage. However, the potential of this extract to protect against NASH under conditions relevant for human pathogenesis remains unclear. Therefore, we studied the effects of bilberry extract on diet-induced NASH in a translational model for disease. Methods: ApoE*3Leiden mice were fed a Western-type cholesterolcontaining diet without (HC) or with 0.1% (w/w) bilberry extract (HCB) for 20 weeks to study effects on diet-induced NASH. Results: Bilberry extract attenuated HC-induced hepatic steatosis, as observed by decreased macrovesicular hepatocellular lipid accumulation and reduced hepatic cholesteryl-ester content. This anti-steatotic effect was accompanied by local anti-inflammatory effects in the liver, as demonstrated by reduced inflammatory cell clusters and myeloperoxidase-positive neutrophilic cells in HCB. On a molecular level, HC-diet significantly induced hepatic expression of pro-inflammatory genes Tnf-a, F4/80, Ccl2, Mpo, Cxcl1 and Cxcl2 while this induction was less pronounced or completely
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POSTERS quenched (F4/80, Mpo, Cxcl1) in HCB. A similar quenching effect was observed for HC-induced pro-fibrotic genes, Acta2 and Col1a1. Consistently, induction of collagen deposition by HC (hepatic fibrosis, determined histologically) was attenuated by bilberry extract. Many of the pro-inflammatory and pro-fibrotic parameters positively correlated with intrahepatic free cholesterol levels. HCB significantly reduced these levels, implicating improved cholesterol handling as a potential hepatoprotective mechanism. Conclusions: Bilberry extract attenuates development of NASH, reducing hepatic lipid accumulation, inflammation and fibrosis, possibly mediated by local anti-inflammatory effects associated with reductions in hepatic free cholesterol. P241 GENOME-WIDE DNA METHYLATION PROFILE IN CHINESE PATIENTS WITH NONALCOHOLIC FATTY LIVER DISEASE Q. Pan, F. Shen, R.-N. Zhang, G.-Y. Chen, J.-F. Lu, J.-Y. Wu, Y.-M. Chen, J.-G. Fan. Department of Gastroenterology, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China E-mail: [email protected] Background and Aims: To uncover the DNA methylation profile of nonalcoholic fatty liver disease (NAFLD) in Chinese Han population. Methods: Two groups of Chinese Han population, being normal group (n = 48) and NAFLD group (n = 57), were enrolled from Tian Jin (north China), Shanghai (middle China) and Zhang Zhou (south China). The genome-wide methylation status of CpG regions in 2 groups were analyzed by peripheral leukocytes using Infinium Human Methylation 450 Bead Chips. The obtained results were then subjected to bio-information analysis on the basis of gene ontology (GO) and signal pathway detection. Results: A total of 2084 differentially methylated loci had been identified between Chinese Han population with and without NAFLD. Of these NAFLD-related loci, 674 loci (32.34%) were located in the CpG islands (CGIs). Of these CGI-located loci, 232 loci were distributed in the promoter regions (34.42%). Signal pathways concerning lipid metabolism and insulin sensitivity, such as Insulin signaling pathway (P = 0.000), Metabolic pathways (P = 0.000), and Adipocytokine signaling pathway (P = 0.001), etc., had been identified to underlie the DNA methylation profile. A lot of differentially regulated GOs, especially enzyme regulator activity (P = 0.000) and lipid binding (P = 0.001), etc., reflected the comprehensive action of genome-wide DNA methylation. Conclusions: The occurrence of NAFLD is correlated with genomewide DNA methylation in CGIs and gene promoters. Altered activity of metabolism-related signal pathways and GOs may reflect the global effect of DNA methylation. P242 METFORMIN DECREASES THE TLR4 EXPRESSION ON PERIPHERAL BLOOD MONOCYTES OF PATIENTS WITH NAFLD A. Zwolak1 , J. Daniluk1 , O. Slabczynska2 , J. Semeniuk2 , M. KandeferSzerszen2 , A. Szuster-Ciesielska2 . 1 Department of Internal Medicine and Internal Medicine in Nursing, Medical University of Lublin, 2 Department of Virology and Immunology, Maria Curie-Skłodowska University, Lublin, Poland E-mail: [email protected] Background and Aims: NAFLD is common cause of chronic liver disease. Increased monocytes expression of Toll-like receptor 4 (TLR4) may be involved in the pathogenesis of NAFLD. The aim of our study was to determine metformin influence on TLR4 expression on monocytes of newly NAFLD diagnosed patients. Methods: The 40 patients were divided into three groups based on their clinicopathological features: i. NASH group with central obesity, ii. simple NAFLD with obesity, S146
iii. simple NAFLD with normal BMI. The next two groups were controls: iv. healthy obese and v. healthy slim persons. The percentage of monocytes expressed TLR4 receptor was estimated by flow cytometry after 24 hour incubation of blood samples with/without metformin (20 or 100mM) or LPS (100 ng/ml). Results: In comparison to blood monocytes of healthy slim persons the expression of TLR4 receptors was significantly higher in all NAFLD patients and correlates with BMI value. Additional in vitro activation of monocytes with LPS increased TLR4 expression only in healthy slim and obese subjects. Supplementation with 100mM of metformin decreased TLR4 expression mainly on monocytes of obese NAFLD/NASH patients while in healthy slim and obese persons expression of TLR4 remained unchanged. When monocytes were simultaneously incubated with LPS and 100mM metformin the reduction of TLR4 expression inversely correlated with disease progress: the lowest decrease appeared in NASH patients, the highest – in healthy slim subjects. Conclusions: Additional beneficial effect of metformin administration could be attenuate of blood monocytes TLR4 expression particularly in early stage of NAFLD. P243 INVOLVEMENT OF MYOKINES AND ADIPOKINES IN LIVER STEATOSIS M. Nakamuta1 , M. Kohjima1 , T. Yoshimoto1 , T. Nakamura1 , T. Ohashi1 , K. Fukuizumi1 , N. Fujimori1 , K. Kawabe1 , K. Haraguchi1 , A. Aso1 , Y. Sumida1 , N. Harada1 , T. Ryu2 , Y. Wada2 , Y. Takami2 , H. Saitsu2 , T. Utsunomiya3 , M. Shimada3 , K. Dohmen4 , H. Nomura5 , M. Enjoji6 . 1 Gastroenterology, Clinical Research Center, 2 HepatoBiliary-Pancreatic Surgery, Clinical Research Center, Kyushu Medical Center, Fukuoka, 3 Digestive and Pediatric Surgery, Tokushima University, Tokushima, 4 Internal Medicine, Chihaya Hospital, Fukuoka, 5 Internal Medicine, Shin-Kokura Hospital, Kitakyushu, 6 Clinical Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Fukuoka, Japan E-mail: [email protected] Background and Aims: Nonalcoholic fatty liver disease (NAFLD) is often accompanied with obesity, hyperlipidemia, diabetes, and/or, insulin resistance, which are simultaneously affect multiple organs. Multiple organ relationship such as between visceral fat, subcutaneous fat, muscle, and liver could be a key for lipid accumulation in the hepatocyte. Here we focused on myokines and adipokines and examined the organ crosstalk between multiple tissues. Methods: We collected visceral fat, subcutaneous fat, muscle, and liver tissue from surgery patients granted permission (n = 139) and extracted mRNA from the tissue. HepG2 or C2C12 cells were cultured in medium supplemented with cytokines or cytokine inhibitors. Expression of the genes was analyzed using quantitative PCR analysis. All experimental procedures were carried out under a protocol approved by ethical committee at Kyushu Medical Center. Results: Expression of Wnt2a in both visceral fat and subcutaneous fat were strongly associated with the expression of ACC1 and CD36 in the liver. FGF2 expression in rectus muscle was also significantly correlated with FAS, SREBP1c and CD36 expression in the liver. Patients with liver steatosis had higher serum FGF2 level than patients without steatosis, and serum FGF2 correlated significantly with Wnt2a mRNA expression in visceral and subcutaneous fat. In HepG2 cells, FGF2 showed stimulative effect on the hepatic fatty acid synthesis, and Wnt inhibitor IWP2 suppressed the expression of FGF2 in C2C12 cells. Conclusions: It is possible muscle FGF2 is secreted from muscle cells stimulated by Wnt2a from adipose tissue, and directly promotes fatty acid synthesis in hepatocyte.
Journal of Hepatology 2014 vol. 60 | S67–S214
P239 METABOLIC SYNDROME AND NON-ALCOHOLIC STEATOHEPATITIS (NASH) ARE ATTENUATED BY INTERVENTION WITH CASPASE-1 INHIBITOR IN HIGH FAT DIET FED LDLR−/− .LEIDEN MICE P.Y. Wielinga, K. Salic, W. Liang, M. Morrison, P. Mulder, T. Kooistra, R. Kleemann. Metabolic Health Research, The Dutch Organization for Applied Scientific Research (TNO), Leiden, Netherlands E-mail: [email protected] Background and Aims: Non-alcoholic steatohepatitis (NASH) is a serious liver pathology which develops as a complication of the metabolic syndrome. Recently, the inflammasome was proposed to be involved in the development of NASH. Here we investigate whether a chemical inhibitor of caspase-1 in already manifest metabolic syndrome would prevent the development to NASH. Methods: Male LDLR−/− .Leiden mice were fed a high fat diet (HFD; group 1) or low fat diet (LFD; group 2) for 21 weeks. In a third group, intervention with caspase-1 inhibitor Ac-YVADCMK (40 mg/kg daily) was started after 9 weeks of HFD (manifest metabolic syndrome) and continued until 21 weeks. Results: HFD treated mice developed obesity and insulin resistance after already 9 weeks of HFD feeding. Intervention with caspase-1 inhibitor attenuated a further development of insulin resistance, and reduced body weight gain as well as adipose tissue inflammation compared to HFD. Histopathological analysis of the livers clearly demonstrated prevention of NASH development with caspase-1 inhibitor, livers were less steatotic and neutrophil infiltration was diminished. Additionally, hepatic fibrosis quantified by sirius red staining and acta2 and col1a1 gene expression as observed in HFD treated mice, was completely prevented. Conclusions: Intervention with a caspase-1 inhibitor in already established disease improved hallmarks of the metabolic syndrome and prevented the development of NASH. Our data further support the importance of caspase-1/inflammasome in the development of NASH and demonstrate that therapeutic intervention in the already ongoing disease process is feasible. P240 BILBERRY EXTRACT ATTENUATES DEVELOPMENT OF NONALCOHOLIC STEATOHEPATITIS IN ApoE3L MICE M.C. Morrison1,2,3 , W. Liang1 , K. Salic1 , P. Mulder1 , P. Wielinga1 , T. Kooistra1 , P. Heeringa2 , R. Kleemann1 . 1 Department of Metabolic Health Research, Netherlands Organization for Applied Scientific Research (TNO), Leiden, 2 Department of Pathology and Medical Biology, University Medical Center Groningen, Groningen, 3 Top Institute Food and Nutrition, Wageningen, Netherlands E-mail: [email protected] Background and Aims: Anthocyanin-rich extract of bilberry fruits may have beneficial effects on lipid metabolism and inflammation and was demonstrated to have hepatoprotective properties in models of restraint-stress- and chemically-induced liver damage. However, the potential of this extract to protect against NASH under conditions relevant for human pathogenesis remains unclear. Therefore, we studied the effects of bilberry extract on diet-induced NASH in a translational model for disease. Methods: ApoE*3Leiden mice were fed a Western-type cholesterolcontaining diet without (HC) or with 0.1% (w/w) bilberry extract (HCB) for 20 weeks to study effects on diet-induced NASH. Results: Bilberry extract attenuated HC-induced hepatic steatosis, as observed by decreased macrovesicular hepatocellular lipid accumulation and reduced hepatic cholesteryl-ester content. This anti-steatotic effect was accompanied by local anti-inflammatory effects in the liver, as demonstrated by reduced inflammatory cell clusters and myeloperoxidase-positive neutrophilic cells in HCB. On a molecular level, HC-diet significantly induced hepatic expression of pro-inflammatory genes Tnf-a, F4/80, Ccl2, Mpo, Cxcl1 and Cxcl2 while this induction was less pronounced or completely
Journal of Hepatology 2014 vol. 60 | S67–S214
S145
POSTERS quenched (F4/80, Mpo, Cxcl1) in HCB. A similar quenching effect was observed for HC-induced pro-fibrotic genes, Acta2 and Col1a1. Consistently, induction of collagen deposition by HC (hepatic fibrosis, determined histologically) was attenuated by bilberry extract. Many of the pro-inflammatory and pro-fibrotic parameters positively correlated with intrahepatic free cholesterol levels. HCB significantly reduced these levels, implicating improved cholesterol handling as a potential hepatoprotective mechanism. Conclusions: Bilberry extract attenuates development of NASH, reducing hepatic lipid accumulation, inflammation and fibrosis, possibly mediated by local anti-inflammatory effects associated with reductions in hepatic free cholesterol. P241 GENOME-WIDE DNA METHYLATION PROFILE IN CHINESE PATIENTS WITH NONALCOHOLIC FATTY LIVER DISEASE Q. Pan, F. Shen, R.-N. Zhang, G.-Y. Chen, J.-F. Lu, J.-Y. Wu, Y.-M. Chen, J.-G. Fan. Department of Gastroenterology, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China E-mail: [email protected] Background and Aims: To uncover the DNA methylation profile of nonalcoholic fatty liver disease (NAFLD) in Chinese Han population. Methods: Two groups of Chinese Han population, being normal group (n = 48) and NAFLD group (n = 57), were enrolled from Tian Jin (north China), Shanghai (middle China) and Zhang Zhou (south China). The genome-wide methylation status of CpG regions in 2 groups were analyzed by peripheral leukocytes using Infinium Human Methylation 450 Bead Chips. The obtained results were then subjected to bio-information analysis on the basis of gene ontology (GO) and signal pathway detection. Results: A total of 2084 differentially methylated loci had been identified between Chinese Han population with and without NAFLD. Of these NAFLD-related loci, 674 loci (32.34%) were located in the CpG islands (CGIs). Of these CGI-located loci, 232 loci were distributed in the promoter regions (34.42%). Signal pathways concerning lipid metabolism and insulin sensitivity, such as Insulin signaling pathway (P = 0.000), Metabolic pathways (P = 0.000), and Adipocytokine signaling pathway (P = 0.001), etc., had been identified to underlie the DNA methylation profile. A lot of differentially regulated GOs, especially enzyme regulator activity (P = 0.000) and lipid binding (P = 0.001), etc., reflected the comprehensive action of genome-wide DNA methylation. Conclusions: The occurrence of NAFLD is correlated with genomewide DNA methylation in CGIs and gene promoters. Altered activity of metabolism-related signal pathways and GOs may reflect the global effect of DNA methylation. P242 METFORMIN DECREASES THE TLR4 EXPRESSION ON PERIPHERAL BLOOD MONOCYTES OF PATIENTS WITH NAFLD A. Zwolak1 , J. Daniluk1 , O. Slabczynska2 , J. Semeniuk2 , M. KandeferSzerszen2 , A. Szuster-Ciesielska2 . 1 Department of Internal Medicine and Internal Medicine in Nursing, Medical University of Lublin, 2 Department of Virology and Immunology, Maria Curie-Skłodowska University, Lublin, Poland E-mail: [email protected] Background and Aims: NAFLD is common cause of chronic liver disease. Increased monocytes expression of Toll-like receptor 4 (TLR4) may be involved in the pathogenesis of NAFLD. The aim of our study was to determine metformin influence on TLR4 expression on monocytes of newly NAFLD diagnosed patients. Methods: The 40 patients were divided into three groups based on their clinicopathological features: i. NASH group with central obesity, ii. simple NAFLD with obesity, S146
iii. simple NAFLD with normal BMI. The next two groups were controls: iv. healthy obese and v. healthy slim persons. The percentage of monocytes expressed TLR4 receptor was estimated by flow cytometry after 24 hour incubation of blood samples with/without metformin (20 or 100mM) or LPS (100 ng/ml). Results: In comparison to blood monocytes of healthy slim persons the expression of TLR4 receptors was significantly higher in all NAFLD patients and correlates with BMI value. Additional in vitro activation of monocytes with LPS increased TLR4 expression only in healthy slim and obese subjects. Supplementation with 100mM of metformin decreased TLR4 expression mainly on monocytes of obese NAFLD/NASH patients while in healthy slim and obese persons expression of TLR4 remained unchanged. When monocytes were simultaneously incubated with LPS and 100mM metformin the reduction of TLR4 expression inversely correlated with disease progress: the lowest decrease appeared in NASH patients, the highest – in healthy slim subjects. Conclusions: Additional beneficial effect of metformin administration could be attenuate of blood monocytes TLR4 expression particularly in early stage of NAFLD. P243 INVOLVEMENT OF MYOKINES AND ADIPOKINES IN LIVER STEATOSIS M. Nakamuta1 , M. Kohjima1 , T. Yoshimoto1 , T. Nakamura1 , T. Ohashi1 , K. Fukuizumi1 , N. Fujimori1 , K. Kawabe1 , K. Haraguchi1 , A. Aso1 , Y. Sumida1 , N. Harada1 , T. Ryu2 , Y. Wada2 , Y. Takami2 , H. Saitsu2 , T. Utsunomiya3 , M. Shimada3 , K. Dohmen4 , H. Nomura5 , M. Enjoji6 . 1 Gastroenterology, Clinical Research Center, 2 HepatoBiliary-Pancreatic Surgery, Clinical Research Center, Kyushu Medical Center, Fukuoka, 3 Digestive and Pediatric Surgery, Tokushima University, Tokushima, 4 Internal Medicine, Chihaya Hospital, Fukuoka, 5 Internal Medicine, Shin-Kokura Hospital, Kitakyushu, 6 Clinical Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Fukuoka, Japan E-mail: [email protected] Background and Aims: Nonalcoholic fatty liver disease (NAFLD) is often accompanied with obesity, hyperlipidemia, diabetes, and/or, insulin resistance, which are simultaneously affect multiple organs. Multiple organ relationship such as between visceral fat, subcutaneous fat, muscle, and liver could be a key for lipid accumulation in the hepatocyte. Here we focused on myokines and adipokines and examined the organ crosstalk between multiple tissues. Methods: We collected visceral fat, subcutaneous fat, muscle, and liver tissue from surgery patients granted permission (n = 139) and extracted mRNA from the tissue. HepG2 or C2C12 cells were cultured in medium supplemented with cytokines or cytokine inhibitors. Expression of the genes was analyzed using quantitative PCR analysis. All experimental procedures were carried out under a protocol approved by ethical committee at Kyushu Medical Center. Results: Expression of Wnt2a in both visceral fat and subcutaneous fat were strongly associated with the expression of ACC1 and CD36 in the liver. FGF2 expression in rectus muscle was also significantly correlated with FAS, SREBP1c and CD36 expression in the liver. Patients with liver steatosis had higher serum FGF2 level than patients without steatosis, and serum FGF2 correlated significantly with Wnt2a mRNA expression in visceral and subcutaneous fat. In HepG2 cells, FGF2 showed stimulative effect on the hepatic fatty acid synthesis, and Wnt inhibitor IWP2 suppressed the expression of FGF2 in C2C12 cells. Conclusions: It is possible muscle FGF2 is secreted from muscle cells stimulated by Wnt2a from adipose tissue, and directly promotes fatty acid synthesis in hepatocyte.
Journal of Hepatology 2014 vol. 60 | S67–S214