Keisuke Yokohama

Sitagliptin can inhibit the development of hepatic steatosis in high-fructose diet-fed ob/ob mice.

The beneficial effect of dipeptidyl peptidase-4 inhibition on diet-induced extra-pancreatic effects, especially on liver tissue remains poorly understood. Thus, we made the experimental designs as follows; five-week-old male ob/ob mice, which develop type 2 diabetic mellitus and nonalcoholic fatty liver disease by taking a high-carbohydrate diet (HCD), were divided into a group in which a HCD was given for 8 weeks as control, and another in which a HCD added with 0.0018% sitagliptin was given for 8 weeks. Hepatic steatosis was seen in all mice, but the mean grade of steatosis in the sitagliptin-administrated mice was significantly decreased. The acetyl-CoA concentrations were lower in sitagliptin-administrated mice, although the differences were not significant. However, the malonyl-CoA concentrations were significantly lower in sitagliptin-administrated mice. The expression of acetyl-CoA carboxylase 1 was inhibited in sitagliptin-administrated mice, irrespective of expressions of carbohydrate responsive element-binding protein (ChREBP) or sterol regulatory element-binding protein (SREBP)-1c. In conclusion, sitagliptin may affect the development of nonalcoholic fatty liver disease by inhibiting the production of malonyl-CoA and thus synthesis of fatty acids in the liver.

Co-Administration of Saireito Enabled the Withdrawal of Corticosteroids in an Elderly Woman with Autoimmune Hepatitis.

An 82-year-old woman with autoimmune hepatitis had been treated with 5 mg prednisolone (PSL) orally to maintain normal transaminase levels. The subsequent transaminase levels were elevated and remained unchanged despite increasing the dose of PSL to 20 mg and introducing ursodeoxycholic acid (UDCA) at a dose of 900 mg. This combined treatment with UDCA was, however, ineffective. Treatment with Saireito at a dose of 9.0 g in conjunction with PSL was then initiated, which led to the subsequent normalization of her transaminase levels. Oral administration of PSL was discontinued eight weeks after the co-administration of Saireito. The patient had a significant response to this treatment.

The preventive effect of the impaired liver function for antiemetic therapy against chemotherapy-induced nausea and vomiting in hepatocellular carcinoma patients

Transarterial chemoembolization and hepatic arterial infusion chemotherapy are recommended for the treatment in patients with intermediate stage of hepatocellular carcinoma. Impaired liver function was sometime observed in patients with hepatocellular carcinoma after transarterial chemoembolization or hepatic arterial infusion chemotherapy. However, what kinds of factors deeply influence in impaired liver function are not clear. A retrospective study was performed to evaluate the risk factors of impaired liver function in cisplatin-naïve patients treated with these therapies using cisplatin. Prior to and 2 months after these therapies, we analyzed the liver function by Child-Pugh score in these patients. For assessing the severity of chemotherapy-induced nausea and vomiting, we utilized the Common Terminology Criteria for Adverse Events ver. 4.0. In hepatocellular carcinoma patients received these therapies using cisplatin, the cancer stage and treatment without neurokinin-1 (NK1) antagonist were found to be independent risk factors of the impaired liver function. The treatment with NK1 antagonist was effective in reducing chemotherapy-induced nausea and vomiting and patients treated with NK1 antagonist kept their liver functions after cisplatin-used these therapies. The treatment with NK1 antagonist was effective in chemotherapy-induced nausea and vomiting and prevented the impaired liver function associated with cisplatin-used these therapies in hepatocellular carcinoma patients.

Effects of Oral L-Carnitine on Liver Functions after Transarterial Chemoembolization in Intermediate-Stage HCC Patients.

Transarterial chemoembolization (TACE) is usually followed by hepatic dysfunction. We evaluated the effects of L-carnitine on post-TACE impaired liver functions. Methods. 53 cirrhotic hepatocellular carcinoma patients at Osaka Medical College were enrolled in this study and assigned into either L-carnitine group receiving 600 mg oral L-carnitine daily or control group. Liver functions were evaluated at pre-TACE and 1, 4, and 12 weeks after TACE. Results. The L-carnitine group maintained Child-Pugh (CP) score at 1 week after TACE and exhibited significant improvement at 4 weeks after TACE (P < 0.01). Conversely, the control group reported a significant CP score deterioration at 1 week (P < 0.05) and 12 weeks after TACE (P < 0.05). L-carnitine suppressed serum albumin deterioration at 1 week after TACE. There were significant differences between L-carnitine and control groups regarding mean serum albumin changes from baseline to 1 week (P < 0.05) and 4 weeks after TACE (P < 0.05). L-carnitine caused prothrombin time improvement from baseline to 1, 4 (P < 0.05), and 12 weeks after TACE. Total bilirubin mean changes from baseline to 1 week after TACE exhibited significant differences between L-carnitine and control groups (P < 0.05). The hepatoprotective effects of L-carnitine were enhanced by branched chain amino acids combination. Conclusion. L-carnitine maintained and improved liver functions after TACE.

Su1811 Sitagliptin Can Be Anti-Inflammatory Drug for the Development of NAFLD in Obese Diabetic OB/OB Mice Under Lipopolysaccharides Existence

the biopsy day. Serum miR-34a was analyzed by quantitative real time PCR method (Applied BiosynthesisTM) and expressed as copies/μL. Results: The mean age of NAFLD patients was 47.1 ± 13.6 year with female of 56%. Their mean body mass index (BMI) was 32.3 ± 16.4 kg/m2. Obesity was found in 74.4%, and metabolic syndrome was 76.4%. Liver histopathology showed that 55.8% of patients had NAS ≥4 and the remaining of 44.2% had NAS <4. Two-third of patients (67.4%) had histopathology compatible with NASH and significant fibrosis ( ≥ F2) was found in 25.6%. Serum level of miR-34a showed significant correlation with NAS (r = 0.44, P = 0.003) (Figure 1), degree of steatosis (r = 0.34, P = 0.026), degree of ballooning (r = 0.31, P = 0.043), and degree of fibrosis (r = 0.34, P = 0.002), whereas miR-34a was not correlated with the degree of lobular inflammation (r = 0.21, P = 0.178). Interestingly, serum miR-34a in patients with NAS ≥4 was significantly higher than those with NAS <4 (338.9 ± 353.2 vs 118.4 ± 154.9, P = 0.01) (Figure 2). There was no significant correlation between serum miR-34a and the other variables including age, body weight, height, and BMI. Conclusion: Serum level of microRNA-34a had significantly fair to good correlation with NAS and degree of fibrosis which represent the severity of inflammation. Additionally, serum miR-34a level in patients with NAS ≥4 was significantly higher than those with NAS <4. Thus, microRNA-34a may serve as a potential biomarker of liver inflammation and fibrosis in NAFLD patients.

Tu1932 Ccl1 Antisense ODN Gene Therapy for Monocytes Is Prevented for the Progression of Hepatocellular Carcinoma in Humanized Chimeric Mice

G A A b st ra ct s death or censored at time of last contact (median follow-up 251 days (d), range 0-1758 d). Survival rates were analysed using the Kaplan-Meier method. Pearsons correlation analyses were performed with respect to survival times and concentrations of IGF-1 and DKK-1. Results. At the time of the analysis 54 patients had died. The concentrations of IGF-1 significantly correlated with survival (0.324, p < 0.01) while the concentration of DKK-1 did not (-0.109, p = 0.197). Survival times using previously published cut-offs for BALAD2 score [-0.91] and IGF-1 [30 ng/ml] are given in table 1. Conclusion. IGF-1 and the BALAD-2 score are valuable to predict survival in patients with hepatocellular carcinoma.

Sa1710 Suppression of Tumor Progression in Patients With Advanced Stage of Hepatocellular Carcinoma by M1 Macrophage Induction

Background: Overcrowding always lead to the shedding of live cells to maintain homeostasis in epithelial cell sheets. Recently, some reports have shown that overcrowded cancer cells can be basally extruded by their neighbors, and then form metastases. However, this interesting phenomenon needs to be further clarified, and the exact mechanisms driving the extrusion of crowded cancer cell sheets remain elusive. We have previously confirmed that BVES, a novel adhesion molecule regulating tight junction formation, could inhibit liver cancer cell extrusion in a simply and skillfully designed 2D cell culture model, as cancer cells are always basally extruded, in this study, we performed a 3D cell culture model to further confirm the inhibition role of BVES in liver cancer cell extrusion. Methods: We designed a 3D cell culture model, after centrifuge, 5×10 6 cells were resuspended by 120μl 10% sucrose solutions, and quickly mixed with equal volume of 0.5% hydrogel solution( BeaverNanoTM ), the cells were then seeded in a Class Bottom Cell Culture Dish(NEST), and allowed to grow for six days. Then, cells were fixed with 4 % paraformaldehyde, incubated with primary antibody and secondary antibody, observed under a confocal microscopy. Extracted cell number were also investigated after enhanced or silenced expression of BVES. RhoA agonist U-46619 and inhibitor C3 enzyme were used to verify the role of RhoA/Rho-kinase pathway in BVES inhibited cancer cell extrusion. Results: Cells successfully grown to form an overcrowded cell mass in the 3D cell culture model, the number of extruded SK-Hep-1 cells, a high metastasis potential cell line, was significantly more than Huh7 cells with low metastasis potential, the normal liver cell line LO2 cells nearly had no cell extruded. Meanwhile, the extruded cells had decreased expression of BVES. Overexpression of BVES inhibited cells extrusion in the 3D cell culture model and increased RhoA activity in SK-Hep-1 cells, while stable knockdown of BVES remarkably promoted cells extrusion and decreased RhoA activity in Huh7 cells. U-46619 increased Huh7 cells extrusion while treated with C3 enzyme significantly diminished cells extrusion. Conclusion: These results suggest that in crowding liver cancer, BVES inactivate RhoA signaling pathway to decrease liver cancer cells extrusion, subsequently may inhibits tumor metastasis. (This study is supported by the National Natural Science Foundation of China NO:81472311 and the Fundamental Research Funds for the Central Universities 2014ZHYX020)

Sa1840 Effects of Oral L-Carnitine on Liver Functions After Transarterial Chemoembolization (TACE) in Intermediate Stage Hepatocellular Carcinoma (HCC) Patients.

Background :Transarterial chemoembolization (TACE) is the standard treatment for intermediate stage hepatocellular carcinoma (HCC) and usually followed by hepatic dysfunction that limits its efficacy. L-carnitine (4-N-trimethyl ammonium 3-hydroxybutyric acid) is recently studied as hepatoprotective agent. In this study, we evaluated L-carnitine effects against the deterioration in liver functions after TACE. Study design: 53 sequential patients with intermediate stage HCC at Osaka Medical College enrolled to this study. All patients were treated by TACE and assigned into two groups; L-carnitine group (26 patients) who received L-carnitine 300 mg tablet twice daily from 2 weeks before to 12 weeks after TACE and Control group (27 patients) without L-carnitine therapy. Liver functions were evaluated for all patients at 2 weeks before, 1, 4, 12 weeks after TACE. 28 of study patients received branched chain amino acids granules. Results: L-carnitine suppressed deterioration in serum albumin level at 1 week after TACE. There were significant differences between L-carnitine group and control group in mean serum albumin change from baseline to 1 week and 4 weeks after TACE (p < 0.05). L-Carnitine maintained Child-Pugh score at 1 week after TACE and exhibited significant improvement at 4 weeks after TACE (p < 0.01 compared to 1 week after TACE). Conversely, control group reported significant Child-Pugh score deterioration from baseline to 1 week after TACE (p < 0.05) and 12 weeks after TACE (p < 0.05). There were significant differences between L-carnitine and control groups in mean Child-Pugh score change from baseline to 4 weeks (p < 0.05) and 12 weeks after TACE (p < 0.05). Interestingly, L-carnitine displayed improvement in prothrombin time (PT) from baseline to 1 week, 4 weeks (p < 0.05) and 12 weeks after TACE. Contrariwise, PT in control group declined less than baseline along all follow up intervals. There were significant differences between L-carnitine and control groups in PT mean change from baseline to 1 week (p < 0.05) and 4 weeks after TACE (p < 0.05). Total bilirubin in L-carnitine group decreased at 1 week post TACE while in control group, total bilirubin at 1 week significantly increased (p = 0.01). Mean total bilirubin change from baseline to 1 week after TACE reported significant differences between L-carnitine and control groups (p < 0.05). Alanine transaminase and C-reactive protein elevation at 1 week after TACE were suppressed in Lcarnitine group. The hepatoprotective effects of L-carnitine were enhanced by concomitant use of branched chain amino acids. No side effects appeared by L-carnitine administration in all clinical courses. Conclusion: L-carnitine and BCAA combination therapy maintained and improved liver functions after TACE and may be offered as a new liver support tool in HCC patients.

Mo1050 The Liver Protection Effect of L-Carnitine Administration After TACE on Intermediate-Stage Hepatocellular Carcinoma Patients

Background: Hepatocellular (HCC) and Cholangiocellular carcinomas (CCA) are the two most common primary liver malignancies affecting the population worldwide and associated with dismal outcomes. Venous thrombosis is often present in patients with liver malignancies and complicates clinical management. It is not clear whether HCC and CCA are associated with different patterns of venous thrombosis since HCC is usually associated with cirrhosis which by itself promotes hypercoagulability. The true frequency and distribution of venous thrombosis in the patients with HCC and CCA is also poorly described. Aim: The goal of the study was to assess the differences in frequency and distribution of portal and nonportal venous thrombosis (PVT and NPVT) in patients with HCC and CCA. Methods: We conducted the retrospective review of the medical records of the 1185 consecutive patients with pathology proven HCC and 559 with pathology proven CCA seen at a tertiary care medical center (Mayo Clinic, Rochester, MN, USA) between 2000 and 2013. The frequency of the venous thrombosis and baseline demographic characteristics were analyzed. Results: Among patients with HCC, 45.6% (540/1185) had associated venous thrombosis. This was comparable to patients with CCA (35.4%; 198/559). Portal venous thrombosis was present in 31% of patients with HCC and 23.4% of patients with CCA. Non-portal venous thrombosis was observed twice less frequently as compared to portal VT and was present in 14.2% of patients with HCC and 12% of patients with CCA. The average age of patients with HCC was similar in portal and non-portal venous thrombosis groups (60 versus 59.5 years respectively) and was comparable to those patients with HCC who did not have venous thrombosis (62 years). This was also true for the patients with CCA where average age of patients with portal venous thrombosis was 61.5 years and non-portal venous thrombosis 62.5 years as compared to average age of 61 years in patients with CCA without venous thrombosis. Across all patient groups (HCC: PVT; HCC:NPVT; CCA:PVT; and CCA:NPVT) male to female ratio was approximately 1:2. Interestingly, females with non-portal venous thrombosis were slightly younger in the HCC group as compared to the CCA group (59 and 65 years respectively). Conclusions: The frequencies of portal and non-portal venous thrombosis are similar in patients with HCC and CCA making radiological and clinical differentiation of these malignancies based only on the distribution of venous thrombosis difficult. The similarity in the venous thrombotic pattern in HCC and CCA points toward a common pathogenesis, and is not likely to be increased by the underlying cirrhosis in HCC.

Mo1772 Lipopolysaccharides Can Accelerate the Development of Nonalcoholic Fatty Liver Disease, Caused by the Up-Regulation of Lipogenic Gene Expressions

Background/Aim: Obesity is rapidly becoming a pandemic and is associated with increased prevalence of iron deficiency anaemia (IDA). Obese populations have higher circulating levels of leptin in contrast to low concentrations of adiponectin. Hence, it is important to evaluate the dynamic role between adiponectin and leptin in obesity-related IDA. Since leptin shares a number of common biological features with IL-6, a major factor in the development of anaemia of chronic inflammation (ACI), we speculated that leptin and its metabolic counterpart adiponectin might play a role in regulating iron metabolism in the overweight population. Methods: The human hepatoma cells HuH7 and Hep G2 were exposed to IL-6 [10-50ng/ml], leptin [0.1-10μg/ml] and adiponectin [5μg/ml] for 16 hours. mRNA expression was determined using real-time quantitative RT-PCR, protein levels were detected by western blot analysis. Both cell lines were also transfected with a hepcidin promoter-luciferase reporter gene construct to investigate transcriptional regulation of hepcidin. Results: Similar to IL-6 (***p<0.001), leptin causes a significant dose-dependent increase of hepcidin promoter activity (***p<0.001) in both cell lines, leading to elevated mRNA-levels. The activation of transcription factor STAT3 seems to play a crucial role. This was confirmed with specific STAT3-Inhibitor S3I201 [50μM] (***p<0.001). Furthermore, we were able to show for the first time that adiponectin completely abolished leptin effects on hepcidin promoter activation (***p<0.001 in HepG2, *p<0.05 in Huh7). Conclusion: Collectively, these data provide a novel insight into the molecular link between obesity and IDA and further demonstrate that adiponectin has the molecular potential to inhibit leptininduced hepcidin expression.