Investigation of the mechanisms of Genkwa Flos hepatotoxicity by a cell metabolomics strategy combined with serum pharmacology in HL-7702 liver cells

1. To investigate Genkwa Flos hepatotoxicity, a cell metabolomics strategy combined with serum pharmacology was performed on human HL-7702 liver cells in this study.

2. Firstly, cell viability and biochemical indicators were determined and the cell morphology was observed to confirm the cell injury and develop a cell hepatotoxicity model. Then, with the help of cell metabolomics based on UPLC-MS, the Genkwa Flos group samples were completely separated from the blank group samples in the score plots and seven upregulated as well as two down-regulated putative biomarkers in the loading plot were identified and confirmed. Besides, two signal molecules and four enzymes involved in biosynthesis pathway of lysophosphatidylcholine and the sphingosine kinase/sphingosine-1-phosphate pathway were determined to investigate the relationship between Genkwa Flos hepatotoxicity and these two classic pathways. Finally, the metabolic pathways related to specific biomarkers and two classic metabolic pathways were analyzed to explain the possible mechanism of Genkwa Flos hepatotoxicity.

3. Based on the results, lipid peroxidation and oxidative stress, phospholipase A₂/lysophosphatidylcholine pathway, the disturbance of sphingosine-1-phosphate metabolic profile centered on sphingosine kinase/sphingosine-1-phosphate pathway and fatty acid metabolism might be critical participators in the progression of liver injury induced by Genkwa Flos.     

Protection of chlordecone-potentiated carbon tetrachloride hepatotoxicity and lethality by partial hepatectomy

Chlordecone (CD) pretreatment is known to markedly potentiate CCl₄ hepatotoxicity. Previous studies have shown that prior exposure to CD obtunds the increased hepatocellular regeneration and repair observed in non-treated rats challenged with a single, low dose of CCl₄. These observations allowed us to hypothesize that suppression of hepatic regeneration and tissue repair by CD + CCl₄ combination treatment might be involved in this interaction. To test this hypothesis, CCl₄ hepatotoxicity was evaluated in actively regenerating livers using CD-treated (10 ppm in the diet for 15 days), surgically partially hepatectomized (PH) male Sprague-Dawley rats. Rats undergoing no surgical manipulation (CTRL) and sham operation (SH) were included as appropriate controls. Surgical manipulations were conducted on day 15 of the dietary protocol. Based on liver-to-body weight ratios (LW/BW), mitotic indices, hepatic cytochrome P-450 content, and hepatic glutathione (GSH and GSSG) levels, PH-induced hepatocellular regeneration was not affected by pretreatment with CD. Thus, the PH model was considered valid for assessing the effects of CD + CCl₄ combination treatment. CCl₄ (100 l/kg; i.p.) was administered 1, 2, 4 or 7 days after the surgical manipulations. Hepatotoxicity was assessed 24 h later by measuring LW/BW and serum enzymes (SGPT, SGOT and ICD) in all four groups. Hepatic histopathological, histomorphometric and lethal effects were assessed in animals receiving CCl₄ 1 or 7 days after the surgical manipulations. CCl₄-induced increases in LW/BW were observed in CD + PH rats receiving CCl₄ 4 or 7 days post-PH, but not in the 1 or 2 day post-PH groups in which the hepatocellular regeneration was maximal. CCl₄-induced serum enzyme elevations were significantly less in the CD + PH rats as compared to CD + SH. This decrease in the serum enzyme elevations was most prominent in the 1 day post-PH group, where the hepatocellular mitotic activity was most pronounced. CCl₄ lethality, assessed in the 1 day post-surgical manipulation group, was also decreased in the CD + PH rats in comparison to CD + SH rats. Such a protection was not observed in rats receiving CCl₄ 7 days post-PH. These data are consistent with and are supportive of the hypothesis that a suppression of otherwise normally stimulated hepatocellular regeneration following low-dose CCl₄ administration is involved in the marked amplification of CCl₄ toxicity by CD.Abbreviations CD chlordecone - GSH reduced glutathione - GSSG oxidized glutathione - PH partial hepatectomy - SH shamhepatectomy - CTRL control, not surgically manipulated - N normal diet - LW/BW liver weight-to-body weight ratio - SGPT serum glutamic; pyruvic transaminase - SGOT serum glutamic oxaloacetic transaminase - ICD isocitrate dehydrogenase These studies were made possible by a grant from the US Environmental Protection Agency R-811072A preliminary report of these findings was presented at the 70th Annual Meetings of the Federation of American Societies for Experimental Biology at St. Louis, MO (Fed Proc 45: 1051, 1986)A. N. Bell is a Predoctoral Toxicology Trainee and Robert A. Young is a Postdoctoral Trainee supported by Toxicology Training grant from National Institute of Environmental Health Science ES-07045     

Intrahepatic Delivery of Glutathione by Conjugation to Dextran

Glutathione was covalently attached to dextran (T-40) by the CNBr activation method. The compound obtained was a water-soluble powder containing 10 (w/w%) glutathione, which was gradually released from the conjugate in aqueous media. Mice depleted of glutathione by treatment with buthionine sulfoximine, a potent inhibitor of -glutamylcysteine synthetase, exhibited a significant increase in hepatic glutathione level after intravenous injection of the conjugate. In mice given a lethal dose of acetaminophen, the survival rate increased progressively with coadministration of the conjugate, whereas little improvement was found when free glutathione was given. The conjugate maintained the serum transaminase activities at lower level after acetaminophen administration. These findings suggest that the dextran conjugate of glutathione is transported into hepatic cells and is intracellulary hydrolyzed to free form, which protects mice from hepatotoxicity due to acetaminophen.     

Comparison of a traditional paracetamol medication and a new paracetamol/paracetamol-methionine ester combination

Summary The SUR 2647 combination is a sachet formulation containing free paracetamol and its N-acetyl-methionate ester (SUR 2647). In a randomized, single-blind, between-patient study the onset of analgesia, duration and efficacy of the SUR 2647 combination vs paracetamol was investigated in outpatients after oral surgery. One group (n=27) received sachets of SUR 2647 combination 2 b.i.d. (equivalent to 2 g paracetamol ×2) on the day of operation, and one sachet b.i.d. (equivalent to 1 g paracetamol ×2) for the following two days. The other group (n=26) received paracetamol tablets 2 q.i.d. on the day of operation (1 g×4) and one tablet q.i.d. (0.5 g×4) for the following two days. Several objective and subjective assessments, including pain score on a visual analogue scale, were recorded for comparison of the postoperative courses. Median onset of analgesia for both groups was 0.5 h. The duration after SUR 2647 combination was 5.5 h as compared to 2.5 h for paracetamol. Mean pain scores showed that the SUR 2647 combination regime reduced pain significantly more than the paracetamol regime from 0.5 to 3.0 h after initiation of medication. The mean pain scores did not show a significant difference during the remaining observation period. Mild to moderate drowsiness was reported in both treatment groups, but it was more common in subjects given SUR 2647 combination.     

Risks Associated with the Use of Garcinia as a Nutritional Complement to Lose Weight

Nowadays, obesity is one of the great nutritional problems facing public health. The prevalence of this pathology has increased in a worrying way over recent years, currently reaching epidemic proportions. In this context, nutritional supplements are presented as a therapeutic alternative to which more and more people are turning to. Nutritional supplements to lose weight based on the Garcinia plant, specifically on Garcinia cambogia, are commonly used. The active principle of this plant to which these properties have been attributed, is hydroxycitric acid (HCA). The aim of the present review is to gather reported data concerning the effectiveness of nutritional supplements based on Garcinia extracts on weight loss and their possible negative effects. Contradictory results have been observed regarding the effectiveness of the supplements. While statistically significant weight loss was observed in some studies, no changes were found in others. Regarding safety, although Garcinia supplements have been revealed as safe in the vast majority of the studies carried out in animal models and humans, some cases of hepatotoxicity, serotonin toxicity and mania have been reported. In conclusion, the results suggest that Garcinia-based supplements could be effective in short-term weight loss, although the data are not conclusive. In addition, the safety of the complement should be further studied.     

肝毒性中药及其与药性和有效成分的关系——对55种中药肝毒性文献资料的分析报告

目的 :研究有关文献资料中中药肝毒性的报告及其与四气、五味、归经、有效成分之间的关系。方法 :应用文献研究方法 ,对 55种具有肝毒性的中药进行统计归类分析研究。结果 :中药四气、五味、归经及其有效成分与肝毒性有一定相关性。肝毒性主要集中在寒、热类 ,苦、辛类 ,归肝、脾、肾经类和有效成分中碱、甙类药物方面。结论 :中药药性理论及有效成分均与肝毒性有一定的关系 ,应用归类分析的方法对文献资料进行分析研究是中药毒性研究中确实可行的方法。     

Characterizing the influence of structure and route of exposure on the disposition of dithiocarbamates using toluene-3,4-dithiol analysis of blood and urinary carbon disulfide metabolites.

Differences in the toxicities observed for dithiocarbamates have been proposed to result from the influence of nitrogen substitution, oxidation state, and route of exposure. To better characterize the fate of dithiocarbmates in vivoas a function of structure and route of exposure, rats were administered equimolar doses of carbon disulfide (CS2), N-methyldithiocarbamate, pyrrolidine dithiocarbamate, N,N-diethyldithiocarbamate, or disulfiram daily for five days, either po or ip, and sequential blood samples obtained. Protein dithiocarbamates formed by the in vivo release of CS2, parent dithiocarbamate, and protein-bound mixed disulfides were assessed in plasma and hemolysate by measuring toluene trithiocarbonate generated upon treatment with toluene-3, 4-dithiol (TdT). To aid in determining the bioavailability of CS2 from the administered dithiocarbamates, the urinary CS2 metabolites, 2-thiothiazolidine-4-carboxylic acid (TTCA) and 2-thiothiazolidin-4-ylcarbonylglycine (TTCG), were also determined. The levels of TdT-reactive moieties detected depended upon both the compound administered and the route of exposure. Parent dithiocarbamates, with the exception of disulfiram, were eliminated from blood within 24 h; but protein associated TdT-reactive moieties persisted and accumulated with repeated exposure, regardless of the route of exposure. N-Methyldithiocarbamate demonstrated the greatest potential to produce intracellular globin modifications, presumably through its unique ability to generate a methylisothiocyanate metabolite. Urinary excretion of TTCA and TTCG was more sensitive than TdT analysis for detecting dithiocarbamate exposure, but TdT analysis appeared to be a better indicator of in vivo release of CS2 by dithiocarbamates than were urinary CS2 metabolites. These data suggest that CS2 is a more important metabolite, following oral exposure, than are other routes of exposure, e.g., inhalation or dermal. In addition, data also suggest that acid stability, nitrogen substitution, and route of exposure are important factors governing the toxicity observed for a particular dithiocarbamate.     

黄褐毛忍冬和灰毡毛忍冬几种成分对大、小鼠化学性肝损伤的保护作用

目的：研究黄褐毛忍冬和灰毡毛忍冬几种成分对四氯化碳（ＣＣｌ_４）,Ｄ-氨基半乳糖（Ｄ-Ｇａｌ）等肝毒物造成大、小鼠肝损伤的保护作用。方法：大、小鼠分别ｓｃ黄褐毛忍冬和灰毡毛忍冬几种成分后,ｉｐＣＣｌ_４或Ｄ-Ｇａｌ,检测血清谷丙转氨酶（ｓＧＰＴ）、肝脏甘油三酯（ＴＧ）和丙二醛（ＭＤＡ）含量。结果：黄褐毛忍冬和灰毡毛忍冬几种成分均不同程度的降低由ＣＣｌ_４和Ｄ-Ｇａｌ引起的ｓＧＰＴ和ＴＧ的升高,降低ＣＣｌ_４引起的ＭＤＡ升高。结论：黄褐毛忍冬和灰毡毛忍冬几种成分具有降低几种肝毒物引起的肝脏毒性的作用。     

三氧化二砷对急性早幼粒细胞白血病患儿的肝脏毒性

目的 研究治疗剂量三氧化二砷(As2O3)对急性早幼粒细胞白血病(APL)患儿的肝脏毒性.方法 对41例APL患儿静脉滴注As2O3注射液,同时进行肝脏毒性监测:肝功能的动态变化、分析影响因素并观察治疗反应.结果 1.肝脏损害的发生率:发生轻度肝损害8例(19.5%),中度肝损害2例(4.9%),未见重度损害.9例(21.9%)ALT升高,10例(24.4%)AST升高,5例(12.2%)γ谷氨酰转肽酶(γ-GT)升高,多在1～3周发生,第4周以后恢复正常.2.肝功能变化:诱导缓解期所有病例的总蛋白、清蛋白、清蛋白与球蛋白比例、总胆红素、前清蛋白和胆碱酯酶均未见动态性差异变化;ALT在第1周开始明显增高,但恢复较快,持续到第3周恢复正常;AST在第1周开始与γ-GT同时明显增高,持续到第4周恢复正常.3.肝功能损害的影响因素:未见患者的年龄、性别、发病时间、有无肝大和肝功能异常、是否存在DIC与肝毒性的发生存在关系;初诊时血常规及LDH与肝毒性的发生亦未见显著性差异;为了达到完全缓解而应用蒽环类药物者,未见肝毒性增加;短期内As2O3的蓄积量变化与肝毒性的发生未见显著性差异;发生分化综合征者,肝毒性的发生率增多.4.治疗反应:暂时停用As2O3,给予保肝治疗后肝功能恢复,未见因肝衰竭而死亡者.结论 常规剂量As2O3治疗小儿APL肝毒性较轻,且短暂,存在一过性肝功能损害,诱导缓解期第1-3周是重点监测时期,应注意分化综合征发生时可能加重肝毒性.As2O3累积量的变化对小儿肝脏功能近期影响不大,但应动态观察肝功能的变化.     

苍耳子水萃取物对大鼠肝脏毒性作用的实验研究

目的：观察不同浓度苍耳子水萃取物对大鼠肝脏功能与形态的影响,为临床安全使用苍耳子提供实验依据。方法：取苍耳子碎粉22 kg,用65%乙醇提取,用石油醚、乙酸乙酯、正丁醇萃取,回收有机溶剂后得到水萃取物743.6 g。水提取物用含3%吐温80生理盐水配制成28.00 mg/mL和1.12 mg/mL混悬液。144只SPF级雄性SD大鼠,体重（200±10）g,用随机数字表法分为苍耳子水萃取物高剂量组、低剂量组和对照组,每组48只。高、低剂量组分别给予28.00 mg/mL和1.12 mg/mL苍耳子水萃取物混悬液2.5 mL每日2次灌胃,对照组以3%吐温80生理盐水2.5 mL灌胃;均连续28 d。分别于给药开始后7、142、12、8 d和停药后71、4 d观察大鼠被毛、摄食量和活动情况,测量大鼠体重,检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、碱性磷酸酶（AKP）水平,取肝脏计算肝脏指数（肝重/体重×100%）,进行肝脏病理分级和评分。结果：苍耳子水萃取物高剂量组大鼠给药28 d内先后出现竖毛、脱毛、疲倦少动、摄食量减少、对外界刺激反应缓慢等症状;低剂量组仅少数大鼠出现少动喜卧、精神萎靡等症状。停药14 d后高剂量组大鼠活动量和摄食量均增加,少动喜卧症状减轻,被毛光泽度恢复。给药212、8 d和停药7 d时,高剂量组大鼠体重（[10.5±4.2）（、10.2±3.1）（、12.1±4.5）g]明显低于低剂量组（[15.3±2.1）（、16.7±4.2）（、17.6±3.2）g]和对照组（[18.6±3.4）（、20.5±5.2）（、19.6±2.5）g],血清ALT（[42.8±3.2）（、49.3±5.9）（、43.2±3.2）U/L]和AST（[108.8±11.7）（、119.8±16.3）（、110.5±17.6）U/L]水平均明显高于低剂量组（[33.7±4.60）（、34.8±5.4）（、33.5±4.9）U/L（,94.7±12.6）（、95.4±10.7）（、96.8±12.8）U/L]和对照组（[31.2±4.3）（、32.5±6.3）、（31.7±5.7）U/L（,92.3±16.2）（、92.9±20.3）（、93.7±16.3）U/L;]血清AKP（[197.2±25.7）（、210.4±41.8）（、189.3±17.6）U/L]水平明显高于低剂量组（[174.3±22.6）（、175.3±27.4）（、176.3±22.8）U/L]和对照组[（171.3±25.6）、（172.5±28.7）（、172.8±26.3）U/L;]给药21、28 d和停药7 d时高剂量组肝脏指数（4.2±0.45、.0±0.7、4.9±0.3）明显高于低剂量组（3.4±0.63、.6±0.53、.9±0.6）和对照组（3.0±0.4、3.2±0.3、3.4±0.5）;给药14、21、28d和停药7d时高剂量组大鼠肝脏病理评分中位数分别为（1.0、1.5、3.0、1.5）,明显高于低剂量组（0.2、0.5、0.9、0.5）和对照组（0.1、0.1、0.2、0.1）。以上各指标高剂量组与低剂量组、对照组间差异均有统计学意义（P〈0.05,P〈0.01）;而低剂量组与对照组间差异均无统计学意义（均P〉0.05）。结论：苍耳子水萃取物致大鼠肝损害与其浓度和作用时间有关;大剂量和长时间用药可加重肝损害。