甘草甜素类药对化学损伤人肝细胞的保护作用比较

目的:研究4种(3类)甘草甜素药物对半乳糖胺(D-GalN)和四氯化碳(CCl_4)损伤培养人肝细胞(L-02)的保护作用及差异,筛选出疗效好的药物,指导临床应用.方法:培养L-02,用复方甘草酸单铵(Compound ammonium glycyrrhizin,CAG)、复方甘草酸苷(Compound glycyrrhizin,CG)、甘草酸二铵(Diammonium glycyrrihizinate,DG)和异甘草酸镁(Magnesium isoglycyrr hizinate,MI)分别进行保护,再经D-GalN或CCl_4处理.观察肝细胞生长状态、测定AST及LDH酶活力、测定细胞内谷胱甘肽(GSH)含量.从而评价CAG、CG、DG和MI对D-GalN和CCl_4损伤L-02细胞的保护作用.结果:浓度为1mg/ml时,CAG、CG、DG和MI均能显著抑制D-GalN和CCl_4所致的AST及LDH释放,提高细胞的存活率.其中CAG抑制D-GalN所致的AST效果显著优于CG、DG和MI(P＜0.05);CAG抑制CCl_4所致的AST及LDH释放的效果显著好于DG和MI;CG抑制D-GalN所致的AST和LDH释放效果显著(P＜0.05)好于MI.浓度为1 mg/ml的CAG、CG、DG和MI 4种药物均能显著抑制2种化学损伤细胞内的GSH降低,其中CAG效果最明显(P＜0.05).结论:1 mg/ml的CAG、CG、DG和MI 4种药物对D-GakN和CCl_4致人肝细胞损伤均有保护作用,其机制可能与抑制GSH降低相关.4种甘草甜素药物,CAG保护肝细胞效果最为显著,CG、DG和MI作用依次减低.     

大鼠再生肝细胞对四氯化碳的耐受

本工作采用肝部分切除(PH)后96h 大鼠的离体肝细胞,以细胞存活率、介质中 GPT、GOT 含量作为肝损伤指标,观察其抗四氯化碳损伤作用。在不加四氯化碳条件下,肝细胞温育3h 或5h 后,细胞存活率、漏入介质中GPT、GOT 活性在假手术组和 PH 组间无显著差异。但在有四氯化碳损伤条件下则不同,在温育3h 或5h 后,存活率在假手术组分别降到52.8±5.9%和43.5±5.8%,而 PH 组分别为69.2±5.8%和60.9±3.2%。漏出的 GPT和 GOT 活性在假手术组也明显高于 PH 组(P<0.01)。提示大鼠离体再生肝细胞有明显的抗四氯化碳损伤作用。     

广东相思子中kaikasaponinⅢ和大豆皂甙Ⅰ对于四氯化碳所致大鼠肝细胞转氨酶升高的影响

豆科植物广东相思子 Abrus cantoniensisHance 全草在中国民间作为治疗肝炎的药物,并在临床上得到证实。作者在经过初级培养的大鼠肝细胞中通过四氯化碳诱导肝损伤模型,对该植物抗肝毒活性成分三萜类皂甙kaikasaponin Ⅲ和大豆皂甙Ⅰ进行了研究。通过测定 GPT 和 GOT 的水平,评估了这2个皂甙(甘草甜素为阳性对照药)的抗肝毒活性。测试剂量为10、50、100、200、和500μg/mL,大豆皂甙Ⅰ抑制 GPT 和 GOT 水平的升高,除剂量为500μg/mL 外,其生物活性     

肝刺激因子对实验性急性肝损伤小鼠的保护作用

大鼠肝刺激因子(HSS)是否有保肝作用?这一问题的阐明不仅有理论意义还有潜在的临床意义。本工作采取初断乳大鼠的肝提取HSS,用CCl_4和半乳糖胺损伤小鼠肝脏,结果观察到:(1)HSS明显降低CCl_4和半乳糖胺所致小鼠血清GPT和GOT升高的幅度;(2)HSS的保肝作用具有量效关系;(3)HSS抗CCl_4损伤肝的时程在72h以上;(4)肝组织切片检查表明,HSS可减轻CCl_4和丰乳糖胺损伤肝的程度。上述生化学和组织形态学的指标表明,HSS对实验性急性肝损伤小鼠确有保护作用。     

四氯化碳亚急性中毒引起肝脏损害机理及部位

利用CCl_4亚急性中毒动物模型发现:CCl_4对大鼠亚急性中毒引起肝匀浆MDA含量显著升高,Schiff碱荧光强度显著增强,并伴随肝匀浆GSH含量显著降低,肝匀浆TG含量显著升高,血清SGPT活性显著增高,这三项肝损害指标变化与LPO指标变化存在平行关系。肝线粒体和微粒体MDA含量均显著升高,肝线粒体标志酶SDHase活性和肝微粒体标志酶G-6-Pase活性均显著降低,且分别与各自亚细胞器MDA含量升高存在平行关系。结果表明:CCl_4亚急性中毒引起肝脏损害的机理之一是LPO;CCl_4亚急性中毒引起肝细胞中的线粒体和微粒体发生LPO损害作用。     

食品添加剂亚硫酸钠对HL-7702肝细胞的毒性作用

目的 探讨亚硫酸钠(Na2SO3)对HL-7702正常人肝细胞的损伤作用.方法 经不同浓度(分别为0.039,0.156,0.625,2.5 mmol/L)的Na2SO3染毒24 h后,采用MTT法观察Na2SO3对肝细胞活性的抑制作用和通过测定肝细胞培养上清中乳酸脱氢酶(LDH)活性来反映其对肝细胞膜的损害.结果 ①随着Na2SO3染毒剂量的增加,OD值逐渐降低,肝细胞的活性抑制率逐渐升高;其中0.625 mmol/L和2.5 mmol/L染毒组与阴性对照组相比有统计学意义(P＜0.05).②肝细胞培养上清中的LDH随染毒剂量的增加而呈上升趋势,与对照组相比有显著性差异(P＜0.05),并且有一定的剂量反应关系(r=0.841,P＜0.01).结论 ①Na2SO3对肝细胞的活性有一定的抑制作用;②Na2SO3染毒24 h后使肝细胞培养上清中LDH活性增高,提示其对肝细胞膜可能有一定的损伤作用.     

大鼠肝刺激因子对CCl_4致肝损伤小鼠保护作用的机制研究——Ⅰ.HSS的抗氧化作用

本工作观察了大鼠肝刺激因子(HSS)对CCl_4损伤小鼠后,肝组织和血清脂质过氧化物丙二醛(MDA)及自由基清除剂谷胱甘肽(GSH)含量的影响。结果发现,HSS(0.1mg/g 体重,ip)可使CCl_4(10%,5ml/kg)中毒后48 h,肝组织 MDA 值的升高幅度明显降低(6.23±0.98/nmol/mg 蛋白vs.4.17±1.67nmol/mg 蛋白,P<0.005),但血清MDA值的下降幅度不明显。同时肝组织GSH含量明显高于CCl_4损伤组(3236±472μg/g vs.2007±589μg/g,P<0.001)。上述结果提示,HSS具有抗氧化作用,降低CCl_4自由基对肝细胞膜的损伤作用,保护受损肝脏。     

金疮小草鲜汁抗小鼠急性四氯化碳肝损伤的作用及机制

目的研究金疮小草鲜汁对四氯化碳(CCl_4)所引起的小鼠急性肝损伤的保护作用及其机制。方法 72只昆明小鼠随机分成空白对照组、模型组、阳性药物水林佳组(30 mg/kg)、金疮小草鲜汁高剂量组(40g/kg)、金疮小草鲜汁中剂量组(20 g/kg)、金疮小草鲜汁低剂量组(10 g/kg)6组。每天灌胃给药一次,连续5 d,末次给药1 h后,除空白对照组外,其他各组均腹腔注射CCl_4造成急性肝损伤。采用HE染色观察肝组织病理变化;检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(T-BIL)含量;检测肝组织内丙二醛(MDA)、超氧化物歧化酶(SOD)含量。结果与模型组比较,金疮小草鲜汁各组均显著降低血清ALT活性(P0.05),显著降低MDA含量(P0.05),提高肝组织超氧化物岐化酶(SOD)活性(P0.05);肝组织切片表明各用药组能减轻肝细胞变形、坏死程度,中剂量组减轻最明显。结论金疮小草鲜汁对CCl_4所致小鼠急性肝损伤有保护作用,其机制可能与抗脂质过氧化有关。     

氟烷和安氟醚麻醉对肝脏功能的影响

50例条件基本相同的胸、脑部手术病人,分别施行50％N_20—50％O_2—氟烷或安氟醚吸入麻醉。于麻醉前,手术后第4天及第10天测定血清AKP、GPT、GOT、LDH、γ—GT值。术后第4天,两组GPT、GOT值升高(P<0.05～0.01),与术后发热有关;氟烷组5项血清酶值均升高(P<0.05～0.01),可能与氟烷中间代谢产物引起肝细胞膜通透性的一过性改变有关。对没有原发肝疾患的病人,本组资料提示,氟烷或安氟醚浅麻醉对肝功能没有明显影响。     

健肝汤抗CCI4所致小鼠急性肝损伤作用的研究

目的 观察健肝汤对CCI4所致急性肝损伤模型小鼠的影响。方法 用0．09％CCI4腹腔注射造成小鼠急性肝损伤模型，观察健肝汤对小鼠血清GPT、GOT以及肝组织形态学的影响。结果 健肝汤高、中、低剂量组均可降低小鼠血清GPT;高剂量组可降低血清GOT，光镜下显示高、中、低剂量组与模型组比肝损伤程度均有不同程度减轻，高、中剂量组可见少量新生的肝细胞。结论 健肝汤有明显的降酶及保护肝细胞的作用。     

右旋儿茶素对四氯化碳或半乳糖胺引起原代培养大鼠...

The effects of d-catechin (d-CTC) on carbon tetrachloride- and d-galactosamine (d-Ga1N)-induced cytotoxicity in primary cultured rat hepatocytes were examined. After 1.5 h preincubation, d-CTC was added at doses of 0.1-5.0 mg/ml to culture medium together with 10 mmol/L CCl4 or 5 mmol/L d-GalN, respectively. GOT, GPT and LDH levels were measured 1.5 h after treatment. The results showed that d-CTC at doses of 0.6 to 5.0 mg/ml could protect the hepatocytes against the toxic effects of CCl4 and d-GalN. At the higher doses (2.5-5.0 mg/ml), d-CTC showed weak inhibition of LDH and GPT activities, but these did not influence its anti-hepatotoxic activity.     

UW液、Celsior液和HTK液低温保存生物人工肝用L-02细胞的效果比较

目的 比较UW液、Celsior液和HTK液4℃常规低温保存生物人工肝用L-02细胞的效果。方法 制备好的L-02细胞悬液分以下3组：UW液保存组（UW液组）;Celsior液保存组（CS液组）;HTK液保存组（HTK液组）。各组细胞于4℃低温保存72h后,分别测定细胞存活率及死亡率（流式细胞术）,ALT及LDH释放,尿素合成功能及白蛋白分泌功能。结果 UW液较Celsior液和HTK液显著提高了4℃常规低温保存保存72h的L-02细胞的存活率[(60.05 ± 4.23)% vs (50.12 ± 3.99)%、(44.20 ±4.67)%],均P<0.05;降低了细胞死亡率[(39.95±4.23)%vs (49.88 ±3.99)%、(55.80%±4.67)%], 均P<0.05;抑制了ALT、LDH释放(均P <0.05);更好地维持了L-02细胞尿素合成功能[ (1.03 ± 0.23)mmol/L vs (0.80± 0.14)mmol/L、(0.48± 0.05)mmol/L]和白蛋白分泌功能[(8.36 ±1.38 )mg/L vs (6.41±1.25)mg/L、(5.19±0.41)mg/L), 均P<0.05。结论 同Celsior液和HTK液相比,使用UW液4℃常规低温保存L-02肝细胞可以明显的提高复温后细胞存活率,降低低温损伤引起的ALT、LDH释放,有效的保护肝细胞尿素合成功能和白蛋白分泌功能,但保存时间不应超过72h。     

丙烯腈对胰岛素抵抗大鼠肝细胞的毒性影响及苯乙基异硫氰酸酯的保护作用

目的:探讨胰岛素抵抗大鼠肝(insulin resistance-Buffalo rat liver, IR-BRL)细胞对丙烯腈毒性的易感性及膳食源化学物苯乙基异硫氰酸酯(β-phenylethyl isothiocyanate, PEITC)对其毒性的拮抗效应。方法:以葡萄糖消耗量、胰岛素受体底物-2(insulin receptor substrate-2,IRS-2)蛋白和胞内磷脂酰肌醇-3激酶(phosphatidylinositol-3 kinase, PI3K)蛋白表达量为评价指标,采用50 nmol/L胰岛素诱导处理BRL细胞24 h,构建IR-BRL细胞模型。以2.5～5.0 mmol/L丙烯腈处理IR-BRL细胞12 h, MTT法检测细胞活力和乳酸脱氢酶(lactate dehydrogenase, LDH)漏出率法检测细胞毒性,综合评价IR-BRL细胞对丙烯腈的易感性特征。以1.25～5.0μmol/L PEITC预处理丙烯腈染毒的IR-BRL细胞1 h, MTT法检测细胞活力和LDH漏出率法检测细胞毒性,评价PEITC减弱IR-BRL细胞对丙烯腈的易感效应。结...     

再灌液中Ca~(2+)浓度的变化对心肌缺血再灌注损伤的影响

在大鼠Langendorff灌流模型上观察灌流液含不同浓度Ca~(2+)对心肌缺血再灌注损伤的影响。结果表明:随再灌液中Ca~(2+)浓度的增加(分别为0、1.5、3.0、5.0mmol/L),以心肌组织谷胱甘肽过氧化物酶活力下降、脂质过氧化物(MDA)含量、Ca含量及乳酸脱氢酶(LDH)漏出量增加为指标的组织损伤进行性加重。     

去甲斑蝥素对LPS 诱导的体外培养的肝细胞损伤的保护作用

目的:通过体外观察去甲斑蝥素(NCTD) 对LPS 所致肝细胞损伤和TNF-α 表达的影响,探讨NCTD 的作用及其机制。方法:胶原酶Ⅳ灌流分离培养大鼠肝细胞,将细胞分为对照组、LPS 组、NCTD 组。对照组用无血清DMEM 培养, LPS 组用LPS (40 mg/L) 诱导,NCTD 组用不同浓度NCTD(0.5,1.0,2.5 μg/mL) 与LPS (40 mg/L) 共同作用,各组作用时间均为24 h。MTT 法检测肝细胞的增殖情况、测定培养上清液乳酸脱氢酶(LDH) 含量,ELISA 法检测TNF-α 和IL-6 表达。结果:LPS (40 mg/L) 作用于原代培养大鼠肝细胞24 h 后,与对照组相比,细胞生长抑制率达27%,培养上清液LDH 含量增加20 倍,TNF-α 和IL-6 的表达以及NF-κB DNA 结合活性均明显增加(P<0.05)。给予不同浓度NCTD 后,培养上清液LDH,TNF-α 和IL-6 的含量及NF-κB DNA 结合活性均明显下降,且呈量效关系。结论:NCTD 拮抗LPS 所致的肝细胞损伤,其保护作用的机制可能与其抑制TNF-α 和IL-6 的表达有关。     

Induction of heme oxygenase-1 in human hepatocytes to protect them from ethanol-induced cytotoxicity

We investigated the relationship between ethanol exposure and heme oxygenase (HO-1) in human hepatocytes in order to ascertain if induction of HO-1 can prevent ethanol induced cellular damage. Methods Dose-dependent (25-100 mmol/L) and time-dependent (0-24 h) ethanol exposure were used in the present study. HO-1 mRNA and protein expression were detected by PT-PCR and Western blot respectively. HO-1 activity was indicated by bilirubin and Fe^2 formation. Cytotoxicity was investigated by means of lactate dehydrogenate (LDH) and aspartate transaminase (AST) level in culture supematants, as well as the intracellular formation of malondialdehyde (MDA), cellular glutathione (GSH) status and CYP 2El activity. Results We first demonstrated a dose-dependent response between ethanol exposure and HO-1 mRNA and protein expression in human hepatocytes. We further observed a time-dependent increase of HO-1 mRNA expression using 100 mmol/L ethanol starting 30 minutes after ethanol exposure, reaching its maximum between 3 h and 9 h, Being similar to what had been demonstrated with the mRNA level,increased protein expression started at 6 h after ethanol exposure, and kept continuous elevated over 18 h. In addition, we found that ethanol exposure to hepatocytes markedly increased HO-1 enzyme activity in a time-dependent manner measured as bilirubin and Fez formation in human hepatocytes.Our results clearly showed that ethanol exposure caused a significant increase of LDH, AST, and MDA levels, while the antioxidant GSH was time-dependently reduced. Furthermore, we demonstrated that pre-administration of cobalt protoporphyrin (CoPP) induced HO-1 in human hepatocytes, and prevented an increase of MDA and a decrease of GSH. These effects could be partially reversed by zinc protoporphyfin (ZnPP), an antagonist of HO-1 induction. Conclusion HO-1 expression in cells or organs could lead to new strategies for better prevention and treatment of ethanol-induced oxidative damage in human liver.     

Low-dose Simvastatin Increases Skeletal Muscle Sensitivity to Caffeine and Halothane

Objective To determine whether the myotoxic side effects of statin simvastatin affect skeletal muscle’s sensitivity to caffeine and halothane. Methods Primary cultured neonate rat skeletal myotubes were treated with 0.01-5.0μmol/L simvastatin for 48 hours. MTT was used to evaluate cellular viability. The gross morphology and microstructure of the myotubes were observed with a light and electron microscope, respectively. The intracellular calcium concentrations ([Ca2+]i) at rest and in response to caffeine and halothane were investigated by fluorescence calcium imaging. Data were analyzed byanalysis of variance (ANOVA) test. Results Simvastatin (0.01-5.0μmol/L) decreased myotube viability, changed their morphological features and microstructure, and increased the resting [Ca2+]i in a dose-dependent manner. Simvastatin did not change myotube’s sensitivity to low doses of caffeine (0.625-2.5 mmol/L) or halothane (1.0-5.0 mmol/L). In response to high-dose caffeine (10.0 mmol/L, 20.0 mmol/L) and halothane (20.0 mmol/L, 40.0 mmol/L), myotubes treated with 0.01μmol/L simvastatin showed a significant increase in sensitivity, but those treated with 1.0μmol/L and 5.0μmol/L simvastatin showed a significant decrease. The sarcoplasmic reticulum Ca2+ storage peaked in the myotubes treated with 0.01μmol/L simvastatin, but it decreased when cells were treated with higher doses of simvastatin (0.1-5.0μmol/L). Conclusions The myotoxic side effect of simvastatin was found to change the sensitivity of myotubes in response to high-dose caffeine and halothane. When dose was low, sensitivity increased mainly because of increased Ca2+ content in the sarcoplasmic reticulum, which might explain why some individuals with statin-induced myotoxic symptoms may show positivecaffeine-halothane contracture test results. However, when the dose was high and the damage to the myotubes was severer, sensitivity was lower. It is here supposed that the damage itself might put individuals with statin-induced myotoxic symptoms at greater risks of presenting with rhabdomyolysis during surgery or while under anesthesia.     

氧化苦参碱对四氯化碳损伤大鼠肝细胞的保护作用

目的 :研究氧化苦参碱对肝细胞的保护作用。方法 :采用原代肝细胞培养的方法 ,选用CCl430mmol/L制备肝损伤模型 ,同时加以不同浓度的氧化苦参碱以保护肝细胞 ,培养 12h后以MTT法测细胞存活率 ,并测定培养基中AST、ALT、LDH。结果 :氧化苦参碱对肝细胞的保护呈剂量依赖性 ,氧化苦参碱组细胞存活率明显高于CCl4损伤组 (98.3% /3.2 % ,P <0 .0 5 ) ;其AST、ALT、LDH较CCl4损伤组明显降低 (依次为 6 .13± 0 .4 4 /896 .87±13.2 3、2 1.5± 0 .76 /15 0 2 .13± 35 .80、19.5± 0 .6 8/2 87.38± 5 .79,均P <0 .0 5 )。结论 :氧化苦参碱对CCl4损伤的肝细胞具较好的保护作用 ,存在最佳剂量性关系。     

黄连素对缺血再灌注心肌细胞损伤的保护作用

目的　研究黄连素对新生大鼠心肌细胞缺血再灌注损伤的保护作用。方法　取体外培养的新生大鼠心肌细胞于缺氧 2 4h复氧 1h造成缺血再灌注 ( I/ R)模型 ,观察细胞损伤情况 ;并将黄连素以 1.5× 10 - 6 m ol/ L、1.5× 10 - 5m ol/ L、1.5× 10 - 4 mol/ L 三种浓度分别加入培养基中 ,预处理 2 4h后 ,再置于上述缺氧复氧环境中培养 ,检测以上不同条件下细胞上清液中的乳酸脱氢酶 ( L DH)、丙二醛 ( MDA)、超氧化物歧化酶 ( SOD)含量 ,并检测各组细胞的凋亡率。结果　与正常对照组相比 ,缺血及再灌组细胞上清液中 L DH、MDA含量明显升高 ( P<0 .0 1) ,SOD活力则显著降低 ( P<0 .0 1) ,缺血组和再灌组凋亡率均升高明显 ( P<0 .0 1)。而用黄连素预处理后缺血及再灌组的 L DH、MDA显著低于用药前 ( P<0 .0 1) ,SOD则高于单纯缺血和再灌组 ( P<0 .0 1) ,上述作用在本实验黄连素浓度为 1.5× 10 - 6 m ol/ L～ 1.5× 10 - 4 mol/ L 范围内 ,随浓度升高而更加明显。特别是用黄连素 1.5× 10 - 5mol/ L 浓度预处理后 ,缺血组和再灌组的细胞凋亡率分别是 14.4%和 2 0 % ,分别与用药前 ( 17.4%和 41% )比较有显著性差异 ( P<0 .0 1)。结论　黄连素对缺血再灌心肌细胞有保护作用 ,其作用与浓度有一定依赖关系     

Protective Effects of Dimethyl—4,4‘—Dinmethoxy—5,6,5’,6‘—Dimethylene Dioxybiphenyl—2,2’—Dicarboxylate on Damages of Isolated Rat Hepatocytes Induced by Carbon Tetrachloride and D—galactosam

The protective effect of biphenyl dimethyl dicarboxylate (DDB) on chemically induced damages was studied in isolated suspended rat hepatocytes. The experimental results showed that DDB (200 micrograms/10(6) cells) efficiently protected the hepatocytes against carbon tetrachloride (CCl4 10 mmol.L-1) and D-galactosamine (1 mmol.L-1) induced damages. Membranal lipid peroxidation (malondialdehyde, MDA formation) and glutamic pyruvic transaminase (GPT) release from the hepatocytes were markedly decreased. The damage of the cell surfaces of the hepatocytes were also reduced as seen under a scanning electron microscope (SEM). Pretreatment with DDB (300 mg.kg-1) orally ameliorated the reduction of liver glycogen and blood glucose caused by ip injection of D-galactosamine (800 mg.kg-1) in mice. When normal rats were given DDB 300 mg.kg-1 once daily for 10 d, the free ribosomal protein and RNA in the liver increased significantly. These results indicate that DDB is of beneficial effects on both damaged and normal hepatocytes.