Preparation of active thiol metabolite of clopidogrel by isolated rat liver perfusion / 中国药房

OBJECTIVE To estab lish the pre paration method of clopidogrel active thiol metabolite （CATM），and to provide reference for the synthesis of cis-CATM. METHODS CATM was prepared ，separated and purified with isolated rat liver perfusion and ChromCore 120 C18 preparative column ，using（S）-2-oxo-clopidogrel as substrate. The target compounds were identified by mass spectrometry and nuclear magnetic resonance spectroscopy. The retention time of the active configuration of CATM in the human body （cis-CATM）were compared to confirm the proportion of active configuration in the target product. RESULTS The conversion rate of the target product was 11.71％. The target products were identified as CATM by MS and 1H-NMR. Peak 2-peak 5 of CATM were four stereoisomers. The retention time of them were 21.3，22.3，26.5，27.3 min. The peak area ratios of them were 7.13％，7.23％，63.52％，14.97％，respectively. Based on that retention time of the active configuration of CATM in human body was 26.3 min，the active cis-stereoisomer in the target product CATM accounted for 63.52％. CONCLUSIONS This method is low-cost ，simple，and can prepare CATM with higher active configuration.

Comparison between different perfusion methods for donor liver acquisition in rat liver transplantation model under direct vision of single operator / 器官移植

Objective To establish a rat liver transplantation model under direct vision of single operator and to explore the effect of different perfusion methods on the quality of the donor liver. Methods On the basis of the "two-cuff method" established by Kamada, the operation details were improved to established the rat liver transplantation model. The recipient rats were divided into two groups according to different perfusion methods, group A (perfusion via abdominal aorta) and group B (perfusion via portal vein). The perfusion effect, operation time, operation success rate, postoperative liver function, liver graft pathological manifestations and survival were compared between the two groups. Results There were more residual red blood cells in sinus hepaticus in group B than in group A after perfusion. Both the donor liver perfusion time and donor operation time were longer in group A than those in group B, and the differences were statistically significant (both P < 0.01). The success rate of operation in group A and group B was 77% and 71%, respectively. At 3 d after liver transplantation in rats, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TB) of the rats in the two groups were significantly higher than normal. At 7, 30 d after operation, compared with group A, the levels of ALT, AST and TB in group B were significantly increased, and the differences were statistically significant (all P < 0.01-0.05). The liver pathological examination showed that the degree of inflammatory reaction in the liver and degree of destruction of liver tissue in group B were more severe than those in group A, but there was no significant difference in long-term survival rate between the two groups. Conclusions Although the perfusion time and donor operation time of rat liver transplantation model were slightly prolonged by means of abdominal aorta perfusion, the perfusion effect was better, which can reduce liver tissue damage after operation and restore liver function to normal levels more quickly.

Participation of hepatic α/β-adrenoceptors and AT1 receptors in glucose release and portal hypertensive response induced by adrenaline or angiotensin II

It has been previously demonstrated that the hemodynamic effect induced by angiotensin II (AII) in the liver was completely abolished by losartan while glucose release was partially affected by losartan. Angiotensin II type 1 (AT1) and adrenergic (∝1- and β-) receptors (AR) belong to the G-proteins superfamily, which signaling promote glycogen breakdown and glucose release. Interactive relationship between AR and AT1-R was shown after blockade of these receptors with specific antagonists. The isolated perfused rat liver was used to study hemodynamic and metabolic responses induced by AII and adrenaline (Adr) in the presence of AT1 (losartan) and ∝1-AR and β-AR antagonists (prazosin and propranolol). All antagonists diminished the hemodynamic response induced by Adr. Losartan abolished hemodynamic response induced by AII, and AR antagonists had no effect when used alone. When combined, the antagonists caused a decrease in the hemodynamic response. The metabolic response induced by Adr was mainly mediated by ∝1-AR. A significant decrease in the hemodynamic response induced by Adr caused by losartan confirmed the participation of AT1-R. The metabolic response induced by AII was impaired by propranolol, indicating the participation of β-AR. When both ARs were blocked, the hemodynamic and metabolic responses were impaired in a cumulative effect. These results suggested that both ARs might be responsible for AII effects. This possible cross-talk between β-AR and AT1-R signaling in the hepatocytes has yet to be investigated and should be considered in the design of specific drugs.

The Relationship between CT Perfusion Imaging and TCM Syndrome of Liver Cancer Patients / 浙江中医药大学学报

Objective] To discuss the relationship between CT perfusion imaging and TCM syndrome of liver cancer patients. [Methods]Through testing the Perfusion parameters of Lesions, surrounding liver tissue and liver tissue perfusion in the distance, we may explore the correlation among perfusion parameters, TCM syndrome of primary liver cancer, then explore the phase rule.[Results] There exists significant difference between perfusion parameters and TCM syndrome of liver cancer patients. Hepatic arterial perfusion(HAP), portal venous perfusion(PVP):deficiency syndrome of both liver and kidney yin>syndrome of heat-damp>syndrome of qi stagnation and blood stasis>syndrome of hepatic stagnation and spleen deficiency;Hepatic perfusion index(HPI):syndrome of hepatic stagnation and spleen deficiency>syndrome of qi stagnation and blood stasis>syndrome of heat-damp>deficiency syndrome of both liver and kidney yin. Child-Pugh classification: syndrome of hepatic stagnation and spleen deficiency(5.34 ±1.46),syndrome of qi stagnation and blood stasis(6.82±0.94),syndrome of heat-damp(8.34±1.12),deficiency syndrome of both liver and kidney yin(9.01±1.19).There exists significant difference between Child-Pugh classification and TCM syndrome of liver cancer patients(P<0.05). The result shows that a high positive correlation between AF, PI and Child-Pugh classification(P<0.05);There exists a negative correlation between PF and Child-Pugh classification(P<0.05).[Conclusion] CT perfusion parameters can be used as an objective indicator of middle-late stage of TCM syndrome of liver cancer patients.

Korean red ginseng prevents ethanol-induced hepatotoxicity in isolated perfused rat liver / 대한수의학회지

Alcohol abuse and its medical and social consequences are a major health problem in many areas of the world. Korean red ginseng (KRG) has been traditionally used for the treatment of liver disease. This study was conducted to evaluate the hepatoprotective effects of KRG against hepatotoxicity in Sprague-Dawley rats treated with ethanol (EtOH). Administration of EtOH for 20 days induced significant changes in serum biochemical parameters (aspartate aminotransferase, alanine transaminase, and glucose) accompanied by histological changes in the liver tissue. Treatment with KRG prior to administration of EtOH inhibited the EtOH-induced biochemical and histological changes of the liver. In perfused rat livers, administration of EtOH caused an increase in lactate dehydrogenase (LDH) release into the perfusate and activated the pro-apoptotic Bax protein but inhibited the anti-apoptotic Bcl-2 protein. Pretreatment with KRG prior to administration of EtOH decreased the EtOH-induced LDH release and inhibition of Bcl-2 protein. These results suggest that KRG exerts anti-apoptotic effects and alleviated EtOH-induced liver injury in rats.

Hemangioma hepático subcapsular com realce perilesional: achados de RM / Hepatic subcapsular hemangioma with perilesional enhancement: MRI features

OBJETIVO: Descrever os achados de imagem de RM no hemangioma hepático com realce perilesional. MATERIAIS E MÉTODOS: Realizou-se uma pesquisa no banco de dados da unidade de RM para identificar todos os casos de hemangioma hepático com realce perilesional entre março de 2008 e julho de 2009. Todos os pacientes foram submetidos a RM pré-contraste em imagens ponderadas em T1 e T2 e em imagens dinâmicas após injeção de gadolínio. Características do hemangioma e do realce perilesional foram avaliadas nas imagens de RM. RESULTADOS: Sete hemangiomas em sete pacientes (cinco homens, duas mulheres; faixa etária entre 41-69 anos; média de 57 anos) foram incluídos no presente estudo. O tamanho das lesões variou de 7 a 20 mm (média de 12,4 mm). Na fase dominante arterial hepática, todos os sete hemangiomas mostraram realce perilesional cuneiforme que se atenuou nas imagens dois minutos após injeção de gadolínio. Quatro dessas lesões demonstraram realce capsular adjacente. CONCLUSÃO: Todos os hemangiomas hepáticos com realce perilesional eram lesões capsulares medindo menos que 2 cm. Tal localização sugere que esses hemangiomas podem recrutar vasos capsulares responsáveis pelo realce perilesional.

OBJECTIVE: To describe the MR imaging features of hepatic hemangioma with perilesional enhancement. MATERIALS AND METHODS: A search was performed of the MRI section database to identify all cases of hepatic hemangioma with perilesional enhancement between March 2008 and July 2009. All patients underwent MR examinations including precontrast T1- and T2-weighted images and postgadolinium dynamic images. On MR images, characteristics of the hemangioma and perilesional enhancement were evaluated. RESULTS: Seven hemangiomas in seven patients (five men, two women; age range, 41-69 years; mean, 57 years) were included in this study. Lesion size ranged from 7 to 20 mm (mean, 12.4 mm). On hepatic arterial dominant phase, all seven hemangiomas exhibited wedge-shaped perilesional enhancement which faded on two minutes postgadolinium images. Four of these lesions demonstrated adjacent capsular enhancement. CONCLUSION: All hepatic hemangiomas with perilesional enhancement were < 2 cm capsule-based lesions. This location suggests that these hemangiomas may conscript capsular vessels to account for the perilesional enhancement.

Effect of Extracorporeal Liver Alloperfusion in Treatment of Acute Liver Failure in Pigs / 中国普外基础与临床杂志

Objective To evaluate the feasibility and curative effect of extracorporeal liver perfusion (ECLP) in treatment of acute liver failure (ALF) in pigs. Methods The experiments were carried out in healthy pigs (weight 20-30 kg) under general anesthesia. All of the pigs were randomly divided into 3 groups. ALF model was established by liver blood supply obstructing and portal-systemic shunting. ALF group (n=5): ALF pigs were killed 8 h after establishing. ALF+ECLP group (n=5): ALF pigs were perfused with ECLP for 4 h and killed 8 h after establishing. Normal liver+ECLP group (n=4): normal pigs were dealt with just as ALF+ECLP group. The data of PT, AST, TNF, blood ammonia were collected in all groups. Pathologic changes in liver and brain were detected. Results The levels of PT, AST, TNF, blood ammonia, RBC and HCT in the ALF+ECLP group were lower than those in the ALF group (P

The Effect of Low Dose Nitroglycerin on Hepatic Blood Flow, Real-time Local Liver Perfusion and Oxygen Extraction Ratio Following the Occlusion and Reperfusion of Hepatic Blood Flow in Experimental Dogs / 대한마취과학회지

BACKGROUND: The Pringle maneuver is traditionally used during the hepatectomy to reduce the blood loss. However, there have been no studies about local liver perfusion (LLP) and oxygen extraction ratio (ERO2) following hepatic ischemia and reperfusion. In this study, the changes in hepatic blood flow (HBF), LLP, ERO2 following hepatic ischemia and reperfusion were observed. And the effects of low dose nitroglycerin (NTG) were observed too. METHODS: A total of 14 mongrel dogs were divided into two groups; control group (C, n = 7), NTG administration group (N, n = 7), NTG administration was started 5 minutes before HBF occlusion. After femoral arterial and central venous catheterization, midline abdominal incision was made. Hepatic artery (HA) and portal vein (PV) were exposed to clamp and declamp. And then doppler flowmeter probes were applied on HA and PV to measure their blood flow and a thermal diffusion microprobe was inserted in the liver parenchyme to measure LLP. RESULTS: The HA and PV blood flow, LLP, and ERO2 were not different between two groups. However, HBF more increased compared to the baseline level in N group after reperfusion. In C group, LLP did not recover after reperfusion. The LLP in N group recovered to the baseline level after reperfusion. CONCLUSIONS: In conclusion, it was observed that the HBF increased and LLP recovered to the baseline level after reperfusion by administration of low dose NTG. The use of low dose NTG is safe and effective for hepatectomy.

TGF-?_1-mediated apoptosis of murine hepatocytes / 医学研究生学报

Objective:The aim of this study was to determine the relationships between the effect of TGF-? 1 and apoptosis in normal and cirrhotic murine hepatocytes. Methods: Liver cirrhosis was induced by means of injection with 50% carbon tetrachloride in BALB/c mice. Hepatocytes were isolated an in situ perfusion method with collagenase solution with minor modification. Normal hepatocytes treated with TGF-? 1(5 ng/ml) were stained with fluorescent DNA-binding dyes Hoechst 33342. The apoptosis of hepatocytes was determined by electrophoresis on 1.5% agarose gel to observe DNA ladder. Results:The viability of hepatocytes isolated from mice by in situ two step collagenase digestion was 95.2%. Internucleosomal fragmented DNA ladder was observed using a 1.5% agarose gel in normal hepatocytes treated with TGF-? 1.obvious DNA ladder from cirrhotic hepatocytes was not observed. The percentage of apoptotic rate in TGF-? 1 treated normal hepatocytes were much higher than that of hepatocytes untreated with TGF-? 1. Conclusion: It was revealed that cirrhotic hepatocytes have poor response to apoptosis induced by TGF-? 1.

Liver perfusion technique and its applications in drug study / 中国药理学通报

Liver perfusion technique has been used in drug study for many decades. Liver perfusion has outstanding advantages over other techniques, such as isolated hepatocytes, hepatic cell lines, and hepatocyte membrane vesicles.It is an ideal experimental model used in drug metabolism, drug-drug interactions, and pharmacokinetic studies. Liver perfusion technique, its advantages or disadvantages, and its extensive applications have been reviewed.

The Effect of Low Dose Nitroglycerin on Hemodynamics and Local Liver Perfusion Following an Occlusion and Reperfusion of the Hepatic Artery and Portal Vein in Experimental Dogs / 대한마취과학회지

BACKGROUND: To reduce massive blood loss during a hepatectomy, many anesthesiologists have used the technique of low central venous pressure maintenance by administration of low dose nitroglycerin (NTG) and/or intravenous fluid reduction. However, so far there have been no studies about local liver perfusion (LLP) changes after hepatic artery (HA) or portal vein (PV) reperfusion in patients receiving nitroglycerin administration. In this study, the changes in hemodynamics and LLP following HA and PV reperfusion along with low dose (2micro gram/kg/min) NTG administration in dogs were observed. METHODS: A total of 20 mongrel dogs were divided into four groups; HA occlusion and reperfusion group (H, n = 5), NTG administration group during the reperfusion on H (H-NTG, n = 5), PV occlusion and reperfusion group (P, n = 5), NTG administration group during the reperfusion on P (P-NTG, n = 5). After femoral and pulmonary arterial catheterization, a midline abdominal incision was made. HA and PV were exposed to clamp and declamp. A thermal diffusion microprobe was inserted in the liver parenchyme to measure LLP. RESULTS: The PV blood flow was not changed after HA occlusion, but HA blood flow increased after PV occlusion. The LLP decreased after HA and PV occlusion. The LLP recovered to the baseline level in group H-NTG after HA reperfusion, but the LLP was more increased compared to the baseline level in group H. In group P, the LLP did not recover after PV reperfusion, but the LLP in group P-NTG recovered to the baseline level after PV reperfusion. CONCLUSIONS: In conclusion, it was observed that the LLP recovered to the baseline level by administration of NTG after PV reperfusion. However, the LLP did not increase after HA reperfusion by administration of low dose NTG.

Ca Effects on Synthesis and Secretion of Insulin-like Growth Factor(IGF-I) and IGF-Binding Proteins by the Perfased Rat Liver / 대한내분비학회지

Background: The insulin-like growth factors, IGF-I and

Hemodynamic characteristics of extracellular UTP in the perfused rat liver

Uridine 5'-triphosphate (UTP) is stored in the granules of cells such as platelets and is released into the extracellular space upon cell stimulation. Extracellular UTP is known to influence many biological processes. We investigated the hemodynamic effects of UTP on the perfused rat liver and characterized its receptors. Liver perfusions were performed in a recirculation system under constant pressure (28 cmH2O). The perfusion flow and oxygen consumption rate were measured at 30 second intervals. UTP decreased the perfusion flow and the oxygen consumption rate, dose-dependently. UTP-induced changes were transient and disappeared in about 10 minutes. Suramin (P2-purinergic antagonist, 100 uM) and indomethacin (cyclooxygenase inhibitor, 20 uM) blocked UTP-induced hemodynamic changes significantly. The effects of UTP were also inhibited when Kupffer cells were damaged with treatment of gadolinium chloride (10 mg/kg iv). L-NAME (1 mM), a potent inhibitor of nitric oxide synthase, markedly enhanced and prolonged the contractile response of UTP in the hepatic vessel. These results suggest that UTP acts mainly on suramin-sensitive UTP receptors on the Kupffer cell through prostanoid synthesis. The nitric oxide systems in the endothelium seem to counteract the vasoconstrictile action of UTP in the hepatic circulation.

The Metabolism of Benzidine-based Dye Direct Black 38 in the Isolated Perfused Rat Liver / 대한산업의학회지

Direct Black 38, a kind of benzidine-based azo dye, is widely used as a dye for fabric, leather, cotton, cellulosic material, paper, wool, silk, and so on. Benzidine-based azo dyes are proven as a mutagen and linked to bladder cancer. In 1978, Natonal Institute for Occupational Safety and Health recommended that three widely used benzidine-based dyes (Direct Black 38, Direct Blue 6, and Direct Brown 95) should be treated as carcinogens. In this experiment, metabolism of the benzidine-based dye. Direct Black 38 was examined by using an isolated liver perfusion system. To measure the metbolites of Direct Black 38,/ 8.0 micrometer, 30.5 micrometer and 63,3 micrometer of Direct Black 38 was added into the recirculating perfusate of the isolated perfused rat liver. Samples were collected at 0, 10, 20, 30, 60, 90. 120 minute. They were treated with sep-pak and methanol, and the metabohtes were detected and quantified with high performance liquid chromatography (HPLC). Residual non-reactive dye in the perfusate and liver was reduced to benzidine and then analyzed by HPLC. Detected metabolites of ?Direct

Influence of Guipi Granule on enzyme AST、ALT in liver perfusion / 中成药

AIM: To analyse the effect of Guipi Granule and serum that contains drug on rat's liver. METHODS: Isolated rat's liver perfusion model was used in the experiment for determining AST and ALT content. RESULTS: Middle and low concentration of Guipi Granule (2.5、5.0g/L) didn't cause any chang of enzyme AST、ALT content in isolated rat's liver perfusion liquid. The effect of serum was not different from Guipi Granule groups and control groups. CONCLUSION: The result shows that Guipi Granule doesn't cause damage to the liver cell.