The antifibrogenic effect of etanercept on development of liver cirrhosis induced by thioacetamide in rats

Liver cirrhosis is an elevating cause of morbidity and mortality worldwide. TNF-α/TNF-R1 signal is implicated in progression of many liver diseases. This study provides histological and ultrastructural view that clarifies the effect of etanercept, a TNF-α inhibitor, on development of thioacetamide (TAA)-induced liver cirrhosis and the accompanied hemosiderosis in rats, highlighting the implication and distribution pattern of hepatic TNF-R1. Sixty male albino rats (Rattus norvegicus) were equally randomized into three groups. Group I served as the control. Liver cirrhosis was triggered in the other two groups by intraperitoneal injection of TAA twice a week for five months. Group II received TAA only, while group III subcutaneously injected with etanercept one hour before TAA, along five months. At the end of the experiment, blood was collected for biochemical analysis and livers were excised for histological, immunohistochemical, and electron microscopical preparations. Rats treated with TAA only developed hepatic cirrhosis accompanied by massive deposition of hemosiderin; strong and widespread expression of hepatic TNF-R1 in sinusoidal endothelial cells (SECs), Kupffer cells (KCs), and many hepatocytes; and frequent appearance of fibrogenic, plasma, and mast cells, at the ultrastructural level. By contrast, administration of etanercept diminished the expression of TNF-R1, attenuated the accumulation of collagen and hemosiderin, and preserved the hepatic histoarchitecture. In conclusion, TNF-α signal via TNF-R1 may be implicated in the mechanism of fibrogenesis and the associated hemosiderosis. Etanercept may provide a promising therapeutic approach not only for attenuating the progression of fibrogenesis, but also for hepatic iron overload-associated disorders.     

Effects of suberoylanilide hydroxamic acid on rat cytochrome P450 enzyme activities

Vorinostat (suberoylanilide hydroxamic acid, SAHA) is the first approved histone deacetylase (HDAC) inhibitor for the treatment of cutaneous T-cell lymphoma after progressive disease following two systemic therapies. The rats were randomly divided into SAHA groups (low, medium and high dosage) and control group. The SAHA group rats were given 12.3, 24.5, and 49 mg/kg SAHA, respectively, by continuous intragastric administration for 7 days. The influence of SAHA on the activities of CYP450 isoforms CYP2B6, CYP1A2, CYP2C19, CYP2D6 and CYP2C9 were evaluated by cocktail method, they were responsed by the changes of pharmacokinetic parameters of bupropion, phenacetin, tolbutamide, metroprolol and omeprazole. The five probe drugs were given to rats through intragastric administration, and the plasma concentration were determined by UPLC-MS/MS. The result of SAHA group compared to control group, there were statistical pharmacokinetics difference for bupropion, phenacetin, tolbutamide and metroprolol. Continuous intragastric administration for 7 days may induce the activities of CYP2C19 of rats, inhibit CYP1A2 and slightly inhibit CYP2B6 and CYP2D6 of rats. This may give advising for reasonable drug use after co-used with SAHA. The results indicated that drug co-administrated with SAHA may need dose adjustment. Furthermore, continuous intragastric administration of SAHA for 7 days, liver cell damaged, causing liver cell edema, in liver metabolism process.     

Hepatoprotectivity of Panduratin A against liver damage: In vivo demonstration with a rat model of cirrhosis induced by thioacetamide

This experiment evaluated Panduratin A (PA), a chalcone isolated from Boesenbergia rotunda rhizomes, for its hepatoprotectivity. Rats were subjected to liver damage induced by intra‐peritoneal injection of thioacetamide (TAA). PA was tested first for its acute toxicity and then administered by oral gavage at doses 5, 10, and 50 mg/kg to rats. At the end of the 8^th week, livers from all rats were excised and evaluated ex vivo. Measurements included alkaline phosphatase (AP), alanine transaminase (ALT), aspartate transaminase (AST) and gamma‐glutamyl transferase (GGT), serum platelet‐derived growth factor (PDGF) and transforming growth factor (TGF‐β1), and hepatic metalloproteinase enzyme (MMP‐2) and its inhibitor extracellular matrix protein (TIMP‐1). Oxidative stress was measured by liver malondialdehyde (MDA) and nitrotyrosine levels, urinary 8‐hydroxy 2‐ deoxyguanosine (8‐OH‐dG), and hepatic antioxidant enzyme activities. The immunohistochemistry of TGF‐β₁ was additionally performed. PA revealed safe dose of 250 mg/kg on experimental rats and positive effect on the liver. The results suggested reduced hepatic stellate cells (HSCs) activity as verified from the attenuation of serum PDGF and TGF‐β1, hepatic MMP‐2 and TIMP‐1, and oxidative stress. The extensive data altogether conclude that PA treatment could protect the liver from the progression of cirrhosis through a possible mechanism inhibiting HSCs activity.     

莪术油注射液致66例不良反应文献分析

目的汇集莪术油注射液的不良反应,为临床合理用药提供参考。方法对国内近几年医药期刊报道应用莪术油注射液出现的66例不良反应进行分类统计与分析。结果莪术油注射液致不良反应与给药剂量无关,主要集中发生在用药过程中30分钟以内,且多发生于<20岁和20-40岁年龄组。结论临床医生、药师及药品生产企业应重视莪术油注射液的不良反应。     

Single dose intravenous thioacetamide administration as a model of acute liver damage in rats

Thioacetamide (TAA) has been used extensively in the development of animal models of acute liver injury. Frequently, TAA is administered intraperitoneally to induce liver damage under anaesthesia. However, it is rarely administered by intravenous injection in conscious rats. The experiments in this study were designed to induce acute liver damage by single intravenous injection of TAA (0, 70 and 280 mg/kg) in unrestrained rats. Biochemical parameters and cytokines measured during the 60-h period following TAA administration, included white blood cells (WBC), haemoglobulin (Hb), platelet, aspartate transferase (GOT), alanine transferase (GPT), total bilirubin (TBIL), direct bilirubin (DBI), albumin, ammonia (NH3), r-glutamyl transpeptidase (r-GT), tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Rats were sacrificed by decapitation 60 h after TAA administration and livers were removed immediately for pathology and immunohistochemical (IHC) examination. Another group of rats were sacrificed by decapitation 1, 6 and 24 h after TAA administration and livers were removed immediately for time course change of pathology and IHC examination. TAA significantly increased blood WBC, GOT, GPT, TBIL, DBIL, NH3, r-GT, TNF-α and IL-6 levels but decreased the blood Hb, platelet and albumin level. The levels of histopathological damage in the liver after intravenous TAA administration were also increased with a dose-dependent trend and more increased at 60 h after TAA administration. The levels of inducible nitric oxide synthase (iNOS) and nuclear factor-κB (NF-κB) detected by IHC in the liver after intravenous TAA administration were also increased with a dose-dependent trend and more increased at 1 h after TAA administration. Single intravenous TAA administration without anaesthesia is a restorable animal model which may be used to investigate acute liver damage.     

四氯化碳大鼠肝硬化形成早期的血液流变学改变

目的：探查大鼠四氯化碳肝硬化形成早期红细胞变形指数、全血粘度等多项血液流变学指数的变化。方法：将Wismr大鼠随机分组,肝组织HE和天狼猩红染色,下腔静脉取血检测红细胞变形指数、全血粘度等血液流变学参数。结果：肝硬化7w鼠的肝组织中有大量胶原纤维增生并开始形成假小叶。对照组和肝硬化组间红细胞数和全血粘度的差异不明显;实验4～5w后,肝硬化组的红细胞变形指数、血浆纤维蛋白原和红细胞血红蛋白含量明显较对照组降低。结论：四氯化碳大鼠肝硬化模型的红细胞变形指数等多项血液流变学参数在第5w后出现明显异常,比形态学上出现典型肝硬化表现约提前了2～3w时间,为临床肝硬化的早期诊断治疗提供了实验依据。     

Effect of emotional-painful stress on cytochrome P-450 induction in the rat liver

Laboratory of Bioenergetics, Research Institute of Pharmacology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR, A. V. Val'dman.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 108, No. 7, pp. 48–50, July, 1989.     

The effect of MS-275 on CYP450 isoforms activity in rats by cocktail method

MS-275, is a potent, class I selective histone deacetylase inhibitor currently in clinical trials for the cure of several types of cancer. The influence of MS-275 on the activities of CYP450 isoforms CYP2B6, CYP1A2, CYP2C9, CYP2D6, CYP3A4 and CYP2C19 were evaluated by cocktail method. The rats were randomly divided into MS-275 group (Low, Medium, High) and control group. The MS-275 group rats were given 12.3, 24.5, 49 mg/kg (Low, Medium, High) MS-275 by continuous intragastric administration for 7 days. The six probe drugs were given to rats through intragastric administration, and the plasma concentration were determinated by UPLC-MS/MS. The result of MS-275 group compared to control group, there were statistical pharmacokinetics difference for bupropion, phenacetin, tolbutamide, metroprolol, midazolam and omeprazole. Continuous intragastric administration for 7 days may induce the activities of CYP2B6, CYP1A2, CYP2C9, CYP2D6, CYP3A4 and CYP2C19 of rats, and may induce the hepatocytes apoptosis. This may give advising for reasonable drug use after co-uesd with MS-275.     

水飞蓟素磷脂复合物对硫代乙酰胺所致大鼠急性肝损伤的保护作用

目的 观察水飞蓟素磷脂复合物灌胃对硫代乙酰胺(TAA)所致大鼠急性肝损伤的保护作用。方法实验分7组,即正常对照组、模型对照组、阴性对照组(磷酯组)、阳性对照组(益肝宁组,即水飞蓟素组)、水飞蓟素磷脂复合物小、中、大剂量组。给药28天后,用硫代乙酰胺(TAA,20mg/kg)造模型。测定各组动物血清中AST、ALT、TP、ALB、LN、HA、PⅢP、CⅣ等指标的变化;并称肝重,计算肝脏指数。结果 与益肝宁(142mg/kg水飞蓟素)相比较,水飞蓟素磷脂复合物灌胃(150、300、600mg/kg)可显著降低硫代乙酰胺肝中毒模型肝损伤的严重程度。结论 水飞蓟素磷脂复合物体内保肝疗效比益肝宁为佳。该作用可能与水飞蓟素磷脂复合物比水飞蓟素的生物利用度高有关。     

Protective effects of pentoxifylline on acute liver injury induced by thioacetamide in rats

Pentoxifylline (PTX) is a non-selective phosphodiesterase inhibitor with the effects of antioxidation, anti-inflammation and anti-fibrosis that has been shown to induce damage in liver. The purpose of this study is to investigate the effects and possible mechanisms of PTX on thioacetamide (TAA)-induced acute liver injury in rats. Male Sprague-Dawley (SD) rats were divided into four groups: control, PTX, TAA and PTX+TAA treated groups. Rats were administrated TAA together with or without PTX for a week and sacrificed 24 h after the last intragastric administration of PTX. Histopathological analysis was carried out. The liver function, the indices of oxidative stress including malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) in liver tissues, and pro-inflammatory cytokines expressions were examined. The mRNA level of NF-κB p65 in liver was also determined. PTX significantly attenuated TAA-induced liver injury. The serum transaminase and MDA levels were reduced while the levels of SOD and GSH were increased, as compared with the TAA-treated group. PTX also remarkably suppressed the secretions of pro-inflammatory cytokines and the nuclear factor-κB (NF-κB) activation induced by TAA. In addition, the histopathological analysis showed that the range and degree of liver tissue lesions were improved obviously in PTX treated group. Pentoxifylline could ameliorate the effects of thioacetamide-induced acute liver injury in rats by inhibiting oxidative stress, expressions of pro-inflammatory cytokines and NF-κB activation.     

不同剂量硫代乙酰胺致肝性脑病小鼠肝脏和脑功能变化

目的:探讨硫代乙酰胺(TAA)致肝性脑病(HE)小鼠肝脏和脑功能变化,寻找TAA的合适剂量和理想的观察指标。方法:设立4个TAA剂量组,分别以200、250、300、400mg/(kg.d)剂量的TAA隔24h行腹腔注射,共3次,建立不同剂量TAA致小鼠HE的动物模型,正常对照组腹腔注射生理盐水,每隔24h观察各组生存率变化,第4d进行脑功能评分、旷场实验和高架十字实验,第5d后观察其血氨和肝功指标。结果:(1)不同剂量的TAA致小鼠存活率差异,400mg/(kg.d)致小鼠存活率过低(31.6%);(2)不同剂量TAA致小鼠脑功能评分、旷场实验的总路程和中央活动时间、高架十字实验的开臂滞留时间百分比显著低于对照组(P0.05),但开臂进入次数百分比与对照组无显著差异(P0.05);(3)不同剂量TAA致小鼠血氨、AST和ALT水平显著低于对照组(P0.001),且不同剂量组之间存在显著差异(P0.001)。结论:(1)300mg/(kg.d)剂量TAA隔日腹腔注射3次为诱导小鼠A型HE模型的适宜剂量和方法;(2)TAA诱导A型HE小鼠脑功能异常,伴有自发活动减少和焦虑情绪,脑功能评分、旷场实验和高架十字实验可用于评估脑功能的变化;(3)TAA致肝功能变化呈现剂量依赖性,血氨、AST和ALT可用于评估肝功能的变化。     

肝硬化大鼠肝癌移植瘤模型建立及影像学特点

目的探讨肝硬化大鼠肝癌原位移植瘤模型的制作及影像学特点。方法55只雄性Wistar大鼠通过腹腔注射二乙基亚硝胺（DEN）诱导肝硬化,然后在硬化程度较轻的50只大鼠肝脏上原位种植CBRH．7919瘤块,4周后磁共振检查筛选出成功模型行超声造影及CT增强检查并分析影像学表现。选取3只肿瘤模型行HE染色检查。结果53只大鼠成功建立肝硬化模型,2只大鼠死亡,肝硬化建模成功率为96．36％（53,55）。肝脏原位移植瘤模型成功率为60％（30150）。30只成功模型肝脏内共长出34个肿块,平均直径为10mm,磁共振检查肿瘤呈稍长信号;CT平扫肿瘤呈稍低密度肿块;多普勒超声提示肿瘤血供丰富;超声造影及CT增强检查动脉期明显强化,静脉期强化消退,部分肿瘤中心动脉期及静脉期均未见强化。肿瘤大体解剖肉眼呈类球型或不规则形,切面肿瘤组织呈鱼肉状;HE染色光学显微镜下肿瘤细胞染色深,大小和形态不一,核大深染、异形性明显,核质比例增大,瘤灶周围肝组织纤维增生明显,正常肝小叶结构消失,形成典型的假小叶结构。结论肝硬化大鼠肝癌原位移植瘤模型建模成功率高,具有可控性和可重复性,其特点和病理学表现与人类肝癌相似,适合肝癌实验性治疗及影像诊断等方面的研究。     

Analysis of gene and protein expression of cytochrome P450 and stress-associated molecules in rat liver after spaceflight

As the incidence and length of spaceflight increases, the need to administer pharmacological agents to crew members may become greater. The final goal is to predict the therapeutic or toxic effects of drugs, especially those with a low therapeutic index, during spaceflight. The liver is central to systemic metabolism, therefore, various drugs rely on hepatic metabolism by cytochrome P450 (CYP). The function of individual CYP enzymes is diverse because they are subject to different regulatory mechanisms and substrate specificity. Because spaceflight may be a stressor and influence each CYP expression individually, the purpose of the present study was to measure the expression of 11 CYP genes and protein distribution in the liver of rats after spaceflight, using real-time polymerase chain reaction and immunohistochemistry, respectively. The gene and protein expression of stress-related proteins such as cold-inducible RNA binding protein (Cirp), heat shock protein 70 (HSP70), HSP90 and the p53 tumor suppressor gene, were also determined. CYP4A1 and Cirp gene expression was significantly increased whereas HSP90 and p53 gene expression was significantly decreased in the flight group than in the ground control. Combined with histology, it is concluded that the effects of spaceflight on the liver may be similar to mild cold stress or fasting.     

Effect of a spleen-sparing operation on functional state of liver mitochondria of rats with experimental cirrhosis

Research Institute of Pediatrics, Academy of Medical Sciences of the USSR, Moscow. Central Research Laboratory, Tashkent Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR M. L. Studenikin.) Translated from Byulleten' £Éksperimental'noi Biologii i Meditsiny, Vol. 111, No. 2, pp. 191–193, February, 1991.     

大豆异黄酮对非酒精性脂肪肝病大鼠的作用

目的探讨大豆异黄酮对高脂饮食诱导大鼠非酒精性脂肪肝病模型的干预作用。方法将50只SD大鼠随机分成5组:正常组,模型组,大豆异黄酮低剂量组(30mg/kg/d),大豆异黄酮中剂量组(100mg/kg/d),大豆异黄酮高剂量组(300mg/kg/d)。除正常组外,其余各组以高脂饮食诱导建立大鼠脂肪肝模型,测定大鼠血清中总胆固醇(TC)、甘油三酯(TG)含量,计算肝指数,对肝脏进行病理学检查,观察大豆异黄酮对该模型的影响。结果模型组大鼠血清中甘油三酯(TG)、胆固醇(TC)水平明显高于正常组(P<0.01);与模型组相比,SI高剂量组及中剂量组大鼠血清中TG、TC水平和肝指数水平均明显低于模型组(P<0.05),SI高剂量组及中剂量组大鼠肝组织的病理损伤明显改善。结论大豆异黄酮对高脂饮食诱导的大鼠脂肪肝有明显的干预作用。     

Antifibrotic effect of edaravone in rat liver cirrhosis induced by dimethylnitrosamine

Edaravone (EDA), a newly synthesized free radical scavenger, has shown excellent results in the treatment of stroke. An overproduction of reactive oxygen species (ROS) causing oxidative DNA damage has been accounted as a major factor causing liver injury and fibrosis. Therefore, we examined its effect of EDA in rat model of liver cirrhosis induced by dimethylnitrosamine (DMN). Ten rats (DMN-group) were injected intraperitoneally with DMN (10 μg/g body weight) alone and another ten rats (EDA-group) were injected intraperitoneally with EDA (10 mg/kg body weight) 2 h after being injected with DMN. Both groups underwent their injection regimen three times a week for 4 weeks, after which the rats were sacrificed and their liver tissue sections were stained with Azan–Mallory for quantitative analyses of fibrosis development, using soft imaging and a previously published scoring system. Additionally, these sections were immunohistochemically stained using an antibody against α-smooth muscle actin (α-SMA). The total-bililubin in the EDA-group was found to be lower than that in the DMN-group. Quantitive analysis of liver fibrosis showed that the fibrotic area of the EDA-group was significantly smaller than that of the DMN-group. Additionally, the number of α-SMA positive cells in the EDA-group were significantly lower than that in the DMN-group. This study showed that EDA reduces liver fibrosis in a rat of cirrhosis induced by DMN. These data suggest that the reduction of liver fibrosis by EDA may be induced by the suppression of activated hepatic stellate cells.     

Short-term effects of splenectomy on serum fibrosis indexes in liver cirrhosis patients

Objective: To determine the changing patterns of 4 liver fibrosis markers pre and post splenectomy (combined with pericardial devascularization [PCDV]) and to examine the short-term effects of splenectomy on liver fibrosis. Methods: Four liver fibrosis markers of 39 liver cirrhosis patients were examined pre, immediately post, 2 days post, and 1 week post (15 cases) splenectomy (combined with PCDV). Results: The laminin (LN) level decreased immediately post surgery compared with the preoperative LN level (P < 0.05). The type IV collagen level decreased immediately post surgery compared with that pre surgery (P < 0.05), it significantly increased (P < 0.05) 2 days post surgery and significantly decreased 1 week post surgery (P < 0.05). Hyaluronic acid and the procollagen III N-terminal peptide levels increased significantly 2 days post surgery compared with that pre and immediately post surgery, they significantly decreased 1 week post surgery compared to 2 days post surgery (P < 0.05). Conclusions: In the short-term, the 4 liver fibrosis markers and the FibroScans post splenectomy showed characteristic changes, splenectomy may transiently initiate the degradation process of liver fibrosis.     

Steroid hormone levels in patients with primary biliary cirrhosis of the liver

Central Research Institute of Gastroenterology, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 109, No. 3, pp. 237–239, March, 1990.     

Activity of nucleolar organizers in hepatocytes of rats with cirrhosis of the liver after treatment with bioactive preparations

The therapeutic efficiency of bioactive preparations of the liver and spleen in the treatment of experimental toxic cirrhosis of the liver is shown. Evaluation of the nucleolar organizer activity showed the highest efficiency of treatment with human fetal liver extract. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 142, No. 7, pp. 114–117, July, 2006