人参总甙抗四氯化碳损伤大鼠离体肝细胞的作用

本实验采用大鼠离体肝细胞原代培养２４ｈ并利用ＣＣｌ４造成急性细胞损伤模型，检定人参总甙（ＧＳＳＤ）对肝细胞损伤的影响。结果表明：ＧＳＳＤ可显著降低中毒肝细胞的脂质过氧化物水平。抑制肝细胞脂质过氧化，并降低谷丙转氨酶（ＧＰＴ）和谷草转氨酶（ＧＯＴ）水平，保护脂质膜。ＧＳＳＤ显著促进中毒肝细胞ＲＮＡ和ＤＮＡ的合成，超微结构观察证实ＧＳＳＤ能减轻ＣＣｌ４对肝细胞染色质，线粒体，内质网和核蛋白体的损害，结     

黄芪皂甙抗犬心肌自由基损伤作用

在黄嘌呤－黄嘌呤氧化酶制造的自由基损伤模型上，用引导培养心肌细胞动作电位和离体心脏灌流的方法，发现黄嘌呤－黄嘌呤氧化酶使动作电位的各参数减少，使离体心功能显著下降，而黄芪皂甙可使上述改变向正常方向转化，因此，黄芪皂甙具有抗自由基损伤作用。     

黄芪多糖减轻慢性镉暴露诱导的大鼠肝脏损伤

目的研究黄芪多糖对慢性镉暴露诱导大鼠肝脏损伤的影响。方法将大鼠随机均分为对照组、模型组和黄芪多糖干预组(用腹腔注射氯化镉复制大鼠肝脏损伤模型),5周后处死大鼠取出肝脏。称重法计算大鼠肝指数;比色分析法检测大鼠血清和肝脏谷丙转氨酶(ALT)活性、谷草转氨酶(AST)活性、乳酸脱氢酶(LDH)活性;用电感耦合等离子质谱仪检测肝脏镉(Cd)含量;酶联免疫吸附试验(ELISA)测定大鼠肝脏白细胞介素-2(IL-2)和转化生长因子β1(TGF-β1)的含量;免疫组化法观察大鼠肝脏Bcl-2和Bax的蛋白表达。结果与对照组比较,模型组大鼠肝指数、ALT活性、AST活性、LDH活性、Cd含量、TGF-β1含量和Bax蛋白表达均升高(P<0.05或P<0.01);但模型组大鼠IL-2含量和Bcl-2蛋白表达下降(P<0.01)。与模型组比较,黄芪多糖能显著降低模型组大鼠肝指数、ALT活性、AST活性、LDH活性、Cd含量、TGF-β1含量和Bax蛋白表达(P<0.05或P<0.01);但黄芪多糖能显著提高模型组大鼠IL-2含量和Bcl-2蛋白表达(P<0.01)。结论黄芪多糖可能是通过调控Bcl-2/Bax蛋白表达来减轻镉致大鼠肝细胞氧化应激损伤。     

黄芪有效成分抗PC12细胞缺氧损伤作用的研究

目的探讨黄芪有效成分抗PC12细胞缺氧损伤的作用。方法超声醇水法提取黄芪有效成分,包括醇溶部分的黄酮、皂苷及水溶部分的黄芪多糖。用连二亚硫酸钠(Na2S2O4)诱导PC12细胞建立缺氧损伤模型,然后加入不同剂量的黄芪提取物,以四甲基偶氮唑盐(MTT)比色法及乳酸脱氢酶(LDH)漏出率测定法观察黄芪有效成分抗PC12细胞缺氧损伤作用。结果缺氧损伤可引起PC12细胞发生明显的损伤性变化,MTT测定的吸光度OD值下降,LDH漏出增加;在损伤后黄酮类可减少LDH漏出,增加吸光度OD值,使细胞存活率明显提高,以终浓度4mg/m l的作用最明显,在缺氧损伤后6h内抗损伤作用最佳,而多糖类此作用不明显。结论黄芪有效成分黄酮类有显著抗Na2S2O4诱导PC12细胞缺氧损伤作用。     

内毒素对实验性急性肝功能不全的作用

本文对大肠杆菌内毒素在半乳糖胺引起急性肝功能不全中的作用进行了研究。实验结果表明,内毒素加重半乳糖胺所致的肝细胞坏死,使大鼠血浆尿素氮、胆红素水平升高,凝血酶原时间延长,嗜睡时间提前与昏迷加重。经血浆、脑匀浆游离氨基酸分析提示,内毒素可使脑组织牛磺酸水平增高。上述结果表明,内毒素血症是诱发急性肝功能不全发生发展的重要因素。     

黄芪抗脑衰老作用的研究

目的研究黄芪对快速衰老鼠脑中12个衰老相关基因表达的影响。方法雄性SAM-P/8老年鼠,随机分为对照组和黄芪组,黄芪组每日腹腔注射黄芪注射液,14d后取材,RT-PCR检测。结果黄芪组与对照组比较。Gabarb3、MAPKK4和Sortilin在鼠脑额叶和海马中表达下调,CalcineurinB、MAP2和P35仅在鼠脑海马中表达下调,RAB6A在鼠脑额叶中表达下调,Calmodulin、MAP1B、RAP2A、SCN2B和Synapsin的表达在黄芪组和对照组之间比较没有统计学差异。结论黄芪可下调上述7种基因在衰老鼠脑中的表达发挥抗衰老作用,通过抑制这些基因产物对脑的消极作用抗衰老。     

丹参提取物F抗大鼠胃窦部粘膜再灌注损伤作用的研究

目的：探讨丹参提取物Ｆ（ＤＳＥＦ）抗大鼠胃窦部粘膜再灌注损伤的作用。方法：大鼠随机分为假手术组（ＳＣ）,单纯失血组（ＨＥ）,生理盐水组（ＮＳ）及丹参提取物Ｆ组（ＤＳＥＦ）;ＳＣ组大鼠除未行放血及血液回输外,行所有操作步骤,ＨＥ组大鼠经２０ｍｉｎ的失血性休克后处死,ＮＳ及ＤＥＦ组在休克２０ｍｉｎ后处死。     

肝炎康对D-氨基半乳糖所致大鼠急性肝损伤的保护作用

目的:观察肝炎康对D-氨基半乳糖(D-GalN)所致大鼠急性肝损伤的保护作用。方法:建立D-GalN致大鼠急性肝损伤模型,设正常对照组,模型对照组,云芝多糖阳性对照组和肝炎康大、中、小剂量治疗组。检测大鼠血清中谷丙转氨酶(ALT)、天门冬氨酸转氨酶(AST)活性和肝组织均浆中丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性,并观察肝脏组织病理学变化,以评价肝炎康对肝脏的保护作用。结果:肝炎康能明显降低由D-GalN所致急性肝损伤大鼠血清中ALT、AST活性,降低肝组织中MDA含量,提高SOD活性,并能明显改善肝脏组织病理损伤。结论:肝炎康对D-GalN所致大鼠急性肝损伤具有一定的保护作用。     

黄芪对大鼠急性肾缺血再灌注损伤的保护作用

肾缺血再灌注损伤过程十分复杂 ,氧自由基的产生并介导组织损伤是其中一个重要环节。黄芪具有提高过氧化物歧化酶 (ＳＯＤ)活性、过氧化氢酶活性及降低脂质过氧化物的药理作用。本文观察黄芪注射液在大鼠急性肾缺血再灌注损伤过程中对肾组织ＳＯＤ活性与丙二醛 (ＭＤＡ)含量的变化。1　材料与方法1 1　实验动物与分组 :雄性ＳＤ大鼠 5 0只 ,体重 2 0 0～ 2 5 0ｇ ,随机分正常组 (Ａ组 ) 5只 ,假手术组 (Ｂ组 ) 15只 ,模型组 (缺血再灌注组 ,Ｃ组 ) 15只 ,黄芪组 (Ｄ组 ) 15只。其中黄芪组在实验前 ,每日腹腔注射黄芪注射液 2ｍＬ 0 1ｋｇ体…     

补体活化在D-氨基半乳糖/内毒素诱导的大鼠实验性肝坏死中的作用

目的：探讨补体系统活化在大鼠实验性肝坏死中的作用。方法：应用D-氨基半乳糖／内毒素（Ga1N／LPS）诱导大鼠肝坏死，用血清50％补体溶解法作为评价血清经典途径补体溶血活性而检测血清CH50、光镜下观察肝组织形态学改变和免疫组化法检测肝组织内补体C3沉积。结果：各组大鼠在用药后随着时间延长血清CH50呈不同程度下降，与对照组比较，单用Ga1N后第4小时血清CH50显著下降（71．43％，P〈0、05），2、6、8、10、12和14小时血清CH50降低更显著（P〈0．01），最低为30、5％；单用LPS后2小时的血清CH50下降至正常水平的69．31％（P〈0．01）；用药后4～6小时，血清CH50有回升趋势，但仍低于正常水平，差异显著（P〈0．05）；Ga1N／LPS组血清CH50的变化与Ga1N组相似，用药后2～14小时血清CH50下降均非常显著（P〈0.01）；应用Ga1N／LPS后2～4小时的血清CH50与LPS组比较差异不显著（P〉0．05），但在用药后6～14小时血清的CH50在两组间差异非常显著（P〈0．01）。联合注射Ga1N／LPS后可见肝组织呈亚大块坏死，并观察到C3沉积于Kupffer细胞、肝细胞膜及坏死区；单用Ga1N组可见肝组织损伤轻微，并观察到有单核细胞浸润、肝细胞气球样变及灶性坏死，但未见C3在肝内沉积；单用LPS组的肝组织形态学改变与Ga1N组类似，C3沉积在内皮细胞和Kupffer细胞。结论：血清补体活化和肝内局部补体活化在Ga1N／LPS大鼠急性肝坏死中起重要作用。外源性LPS可引起大鼠血清补体系统经典途径的活化，但其所致的肝损伤可能与肝组织中补体C3的局部沉积有关。Ga1N所致肝损伤可能与血清补体活化无明显关系。     

Effect of phenobarbital on hepatic injury induced by chronic carbon tetrachloride treatment

Rats were given 1 ml CCl4 per kg body weight subcutaneously 2 times a week, for 16 weeks. The effects of simultaneous phenobarbital (PB) treatment (0.05% in drinking water) on the hepatotoxicity of CCl4 was studied during 16 weeks of treatment. The retardation of growth, the increase in liver weight and mortality were greater in animals receiving both PB and CCl4 than those given CCl4 alone. Cirrhosis was apparent only in animals treated by PB + CCl4. The potentiating effect of PB on CCl4 hepatotoxicity was also seen in hexobarbital sleeping time, the rate of hexobarbital metabolism, and the cytochrome P-450 content in liver microsomes. The inducing effect of PB alone decreased with time both in vivo and in vitro, which suggests an adaptation or some kind of exhaustion of liver to the effects of PB.     

细脚拟青霉多糖对抗四氯化碳诱导的急性肝损伤

目的：观察细脚拟青霉粗多糖（cPtPs）及其纯化多糖（PtPs）对四氯化碳（CCl4）诱导大鼠急性肝坏死的影响。方法：将Wistar大鼠分为4组（对照组、CCl4组、cPtPs +CCl4组和PtPs +CCl4组），分别用生理盐水、cPtPs和PtPs灌胃15 d，最后2 d，腹腔注射CCl4，16 h后全自动生化分析仪检测血清中丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、总胆红素（TBIL）、直接胆红素（DBIL）和间接胆红素（IBIL）；肝组织苏木素-伊红（HE）染色观察病变程度；黄嘌呤氧化酶法和硫代巴比妥酸法分别检测肝匀浆中超氧化物歧化酶（SOD）和丙二醛（MDA）；甲基百里香酚蓝比色法测定肝细胞线粒体中Ca2+ 浓度；免疫组织化学法检测肝组织中α-平滑肌肌动蛋白（α-SMA）的表达。结果：与对照组比较，CCl4组血清中ALT、AST、TBIL、DBIL和IBIL的含量显著增加（P＜0.05），HE染色肝组织变性坏死累及全小叶。与CCl4组比较，PtPs+CCl4组血清中ALT、AST、TBIL、DBIL和IBIL的含量显著下降（P＜0.05）；PtPs +CCl4组HE染色病变程度较轻，变性坏死局限于肝小叶的Ⅲ区；PtPs +CCl4组胞浆中SOD活性提高， MDA水平降低（P＜0.05）；PtPs +CCl4组和cPtPs +CCl4组线粒体中Ca2+ 浓度均下降（分别为P＜0.05，P＜0.01）；PtPs +CCl4组α-SMA在肝组织的坏死区几乎无表达。结论：PtPs能显著减轻CCl4诱导的肝损伤，可能与PtPs抗脂质过氧化作用相关，效果优于cPtPs。     

Carbon tetrachloride-induced acute liver injury in Mini and Wistar rats

Acute liver injury induced by CCl4 injection (0.5 ml/kg b.w.) was compared between Mini and Wistar rats. Mini rats (Jcl:Wistar-TgN (ARGHGEN)1Nts strain) are Wistar-derived transgenic animals in which the expression of growth hormone (GH) gene is suppressed by the presence of an antisense transgene. The hepatic lesion appeared earlier and its recovery was delayed in Mini rats compared to in Wistar rats. The degree of the liver injury was more severe in Mini rats than in Wistar rats, and this corresponded well with the changes in serum AST level. Moreover, in accordance with the localization of CYP2E1-positive hepatocytes in the early stage after CCl4 treatment, the initial lesion characterized by ballooning of hepatocytes developed in the centrilobular zone in Wistar rats while it appeared in the middle zone in Mini rats. The changes in the percentage of PCNA-positive cells and the levels of HGF and TGF-beta1 mRNAs were clearly different between the two strains. These results indicate that the response of the liver to CCl4 is different between GH-suppressed Mini rats and Wistar rats.     

四氯化碳致小鼠急性肝损伤的性别差异及其与雄激素受体表达的关系

目的比较CCl4对不同性别小鼠急性肝损伤的诱导作用及其对肝脏雄激素受体(AR)表达的影响。方法健康成年昆明小鼠70只,随机分为CCl4雄性组、CCl4雌性组、雄性对照组,雌性对照组。50%CCl4-粟米油溶液皮下注射,0.6ml/100g体重,1周2次,诱导小鼠急性肝损伤。造模第7天处死各组动物取材固定,常规HE染色比较不同组别小鼠肝组织病理变化;免疫组织化学法检测各组小鼠肝脏AR的表达,根据阳性细胞反应强度及百分比设定0级(阴性)及1-3级(阳性)标准,双盲计分法统计各组AR表达分值。结果实验组CCl4造模过程中雄性组小鼠死亡率为60%(15/25),明显高于雌性组(12%,3/25),对照组小鼠无死亡(0/10,0/10)。肝组织HE染色结果可见CCl4诱导的急性肝损伤主要部位为肝小叶周围的门管区及界板区,损伤部位肝细胞出现脂肪变,少数肝组织出现点状坏死。免疫组化显示雄性及雌性对照组小鼠肝细胞AR呈弱阳性表达,不同性别小鼠肝细胞AR表达差别无统计学意义;实验组各组别肝小叶周围受损肝细胞出现较强的AR表达,其中CCl4雄性组AR表达分值明显高于CCl4雌性组小鼠(P<0.01)。结论雄性小鼠对急性肝损伤的耐受性低于雌性小鼠,其中AR的高表达与CCl4诱导的急性肝损伤性别差异密切相关。     

Hepatobiliary excretion of cysteinyl leukotrienes in three experimental models of acute hepatic injury

The acute phase response to chemically-induced organ damage involves inflammation and the production of leukotrienes. The liver ordinarily takes up, metabolizes and excretes into bile cysteinyl leukotrienes, but the effect of hepatic injury on these processes is unknown. The hepatic uptake and biliary excretion of LTC₄ was studied in male Sprague-Dawley rats after exposure to either streptozotocin (45 mg/kg iv 30 days before experimentation), estradiol-17 β-valerate (1mg/kg sc once a week for 3 weeks) or lipopolysaccharide/d-galactosamine (33 μg/kg ip; 300 mg/kg ip at 6 h and 3 h, respectively, before experimentation). Acute liver injury is produced by these treatment paradigms. Glucose concentrations and activities of several marker enzymes in plasma were measured to demonstrate hepatic injury. Biliary excretion of³H-LTC₄ was similar to normal control rats in the three types of acute liver injury. Bile flow rates after³H-LTC₄ injection were reduced in lipopolysaccharide-pretreated rats and increased in estradiol-treated animals. Total biliary excretion of leukotrienes was not altered in any disease group. Thus, these models of acute hepatic injury do not appear to influence the hepatobiliary clearance of leukotrienes. accepted by G. W. Carter     

High-mobility group box 1 exacerbates CCl4-induced acute liver injury in mice

High-mobility group box 1 (HMGB1) is a nuclear factor that can also serve as an imflammatory mediator once released into extracellular milieu. Therefore, HMGB1 has been recognized to play a pivotal role in inflammatory diseases such as sepsis, acute lung injury, ischemia reperfusion injury and type 1 diabetes. Nevertheless, its impact on carbon tetrachloride (CCl₄)-induced hepatic injury is yet to be elucidated. In the present report, we demonstrated evidence indicating that high levels of HMGB1 were not only present in the necrotic area of liver but also in the serum after CCl₄ challenge. In line with these observations, administration of exogenous recombinant HMGB1 exacerbated CCl₄-induced hepatic injury, while HMGB1 blocking antibody provided protection for mice against CCl₄-induced acute liver injury as evidenced by the decrease of serum transaminase and reduction of hepatic tissues necrosis. Mechanistic studies revealed that blockade of HMGB1 attenuated CCl₄-induced MDA accumulation along with improved SOD and GSH activity. Treatment of mice with HMGB1 neutralizing antibody also significantly inhibited the production of proinflammatory mediators TNF-α and IL-6 along with attenuated HMGB1 expression and its extracellular release. Together, our data suggest an essential role for HMGB1 in CCl₄-induced acute liver injury, while HMGB1 neutralizing antibody could be served as an effective regimen for preventing CCl₄-induced acute liver injury.     

大鼠肝纤维化程度与门脉压力的关系

为证实肝纤维化在肝硬化门脉高压形成中的作用,我们用半自动图象分析仪测定了大鼠CCl_4肝硬化的肝脏胶原含量,同时用检压计测定了同组动物的门脉压力,分析了两者的相关件。结果表明,正常大鼠肝内胶原面积为3206.30±864.62μm~2,肝硬化大鼠为32390.16±11879.98μm~2,差异极显著(P<0.001)。前者与参考面积的比值为1.58±0.43%,后者为15.97±5.86%。正常鼠肝脏胶原面积与门脉压力梯度没有相关性,肝硬化鼠血管周围面积(r=0.44,P<0.05)、肝小叶内胶原面积(r=0.79,P<0.01)及肝内胶原总面积(r=0.76,P<0.01)均与门脉压力梯度呈显著正相关。结果提示,在CCl_4肝硬化大鼠,肝纤维化是导致门脉高压的重要原因之一,支持门脉高压形成的后向血流学说。     

Modulation of perisinusoidal cell cytoskeletal features during experimental hepatic fibrosis

Summary Hepatic perisinusoidal cells (PSCs) proliferate and are thought to be the principal source of extracellular matrix proteins during the development of liver fibrosis. We have studied the classical model of carbon tetrachloride induced liver fibrosis in order to evaluate the possible modulation of PSCs into a synthetically active and contractile cell: the myofibroblast (MF). At the ultrastructural level, this modulation was characterized by reduction of lipid vacuoles and appearance of a developed rough endoplasmic reticulum as well as of microfilament bundles. On investigating the cytoskeletal equipment of PSCs and MFs using light and electron microscopic immunohistochemistry, we found a heterogeneity of phenotypic features. While typical PSCs in normal and fibrotic livers always contained desmin, MFs expressed-smooth muscle (SM) actin in areas of tissue injury and active fibrogenesis. Cells co-expressing-SM actin and desmin were most prominent in the prevenular zone of the lobule (known to be vulnerable to carbon tetrachloride toxicity) and in developing fibrous septa. As demonstrated by immunogold electron microscopy, labelling of microfilament bundles by-SM actin antibody was noted in PSCs containing lipid droplets in early stages of fibrosis; here MFs gradually accumulated and showed-SM actin containing microfilament bundles. In scar tissue,-SM actin expression decreased in both PSCs and myofibroblasts. Our observations support the concept of phenotypic plasticity of PSCs and confirm, at the ultrastructural level, previous suggestions of modulation of these cells into MFs in the course of liver fibrosis.     

缺血预处理通过抑制白三烯C4生成减轻大鼠肝缺血再灌注损伤

 目的：研究缺血预处理（IP）减轻大鼠肝脏缺血再灌注（I/R）损伤是否涉及前炎症因子白三烯C4(LTC4)。方法：健康成年雄性SD大鼠随机分为3组(每组6只)。I/R组： 采用大鼠部分（70%左右）肝脏缺血60 min再灌注5 h模型,缺血前15 min开始至复灌5 h经颈外静脉输注生理盐水（3 mL·kg-1·min-1）;假手术组：只麻醉开腹,不阻断肝脏血流;IP组：在I/R前先阻断肝左、中叶血流10 min,然后开放血流10 min,余步骤同I/R模型组。应用反相高效液相色谱法（RP-HPLC）检查肝组织LTC4含量,同时生化检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）活性和肝组织谷胱甘肽（GSH）含量,以及HE染色法检查肝组织学损伤。结果：肝脏IP处理完全逆转了再灌注5 h所致肝组织LTC4含量增加（P<0.05）;同时增加肝组织GSH含量,明显降低血清ALT和AST活性（P<0.05）,并减轻肝脏组织结构损伤。结论：IP处理减少肝脏I/R期间LTC4堆积,同时伴随血清肝酶释放减少和肝组织结构损伤降低,以及保护肝组织氧化还原状态,表明IP的有利影响可能涉及其在肝脏I/R损伤期间抑制LTC4生成。