Caspase抑制剂F1013对刀豆蛋白A诱导小鼠急性肝损伤的治疗作用

目的研究Caspase抑制剂F1013对刀豆蛋白A(Co-nA)所致小鼠急性肝损伤模型的治疗作用,并对其机制进行初步探讨。方法 60只♂BALB/c小鼠随机分成对照组、模型组、阳性药组(NAC 155 mg.kg-1)和F1013(5,2.5,1.25 mg.kg-1)给药组。除对照组外,其余组小鼠均尾静脉注射ConA建立小鼠急性肝损伤模型,造模后2 h,阳性药组腹腔注射NAC,F1013给药组和模型组分别皮下注射F1013和溶媒。造模后8 h留取血清检测丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素(T-Bil)含量和TNF-α水平,肝组织进行HE染色,并检测肝细胞凋亡率。结果在Co-nA诱导小鼠急性肝损伤模型,F1013组均明显改善肝脏病理组织损伤;明显降低血清ALT、AST、T-Bil和TNF-α水平及肝细胞凋亡率(P<0.01或P<0.05)。结论 F1013对Co-nA所致小鼠急性肝损伤具有较好的治疗作用,其机制可能与减少TNF-α的产生及抗肝细胞凋亡有关。     

Caspase抑制剂F1013对急性肝衰竭大鼠肝细胞凋亡的影响

目的：观察F1013对D-氨基半乳糖（D-GalN）及脂多糖（LPS）所致大鼠急性肝衰竭模型的保护作用,并对其机制进行初步探讨。方法：采用D-GalN/LPS诱导大鼠急性肝衰竭模型,用全自动生化分析仪检测血清ALT、AST和总胆红素（TBIL）含量;用苏木素-伊红（HE）染色,观察肝组织病理学变化;采用流式细胞学技术和原位末端标记（TUNEL）法检测肝细胞凋亡情况。结果：模型组大鼠血清ALT、AST、TBIL水平和肝细胞凋亡率较正常对照组均显著升高（P〈0.01）。F1013（5,2.5,1.25 mg/kg）给药组大鼠肝组织病理损伤程度明显轻于模型组;血清ALT、AST水平和肝细胞凋亡率均较模型组明显降低（P〈0.01或P〈0.05）,另F1013（5 mg/kg）给药组血清TBIL水平较模型组明显降低（P〈0.01）。结论：F1013对GalN/LPS引起的大鼠急性肝衰竭有较好的保护作用,其机制可能与抑制肝细胞凋亡有关。     

前列地尔对D-氨基半乳糖致急性肝损伤大鼠肝匀浆IL-18水平升高的抑制作用

目的:观察前列地尔对D-氨基半乳糖致急性肝损伤大鼠肝匀浆IL-18水平升高的抑制作用。方法:32只SD大鼠一次性腹腔注射D-氨基半乳糖(D-Gal)1.4 g.kg-1,建立急性肝损伤模型。造模后大鼠随机分为前列地尔注射液治疗组(L ipo PGE130μg.kg-1)和阴性对照组(灭菌生理氯化钠溶液),均为一次性腹腔注射。检测两组造模前和造模后6,12,24 h血清谷丙转氨酶(ALT)、谷草转氨酶(AST)和肝匀浆IL-18的水平。结果:D-氨基半乳糖造模后,两组大鼠血清ALT,AST和肝匀浆IL-18值呈动态升高(P<0.05)。在造模后6,12和24 h,前列地尔30μg.kg-1治疗组的血清ALT,AST和肝匀浆IL-18值均明显低于阴性对照组。结论:前列地尔对D-氨基半乳糖致急性肝损伤大鼠有肝脏保护作用,其机制可能与降低肝脏IL-18水平有关。     

蛇葡萄根的保肝降酶及抗自由基损伤作用

用D-氨基半乳糖(D-GalN)制备大鼠急性肝损伤模型,检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)和肝匀浆超氧化物歧化酶(SOD)、丙二醛(MDA)含量,观察肝组织病理变化,以探讨中药蛇葡萄根对急性肝损害的保护作用。结果显示:蛇葡萄很能显著降低血清ALT、AST含量,提高SOD含量,降低MDA含量,高剂量组还能减轻肝细胞受损程度。提示蛇葡萄根具有保肝降酶及抗自由基损伤作用。     

富锗金针菇多糖肝保护有效成分的研究

目的寻找富锗金针菇粗多糖(CFVP)中发挥肝保护作用的有效成分。方法采用0.2%CCl4腹腔注射造成小鼠急性肝损伤模型,以富锗金针菇精品多糖FVP1、FVP2静脉给药,测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)的活力,肝组织匀浆,测定匀浆液中丙二醛(MDA)含量;采用二步分离法制备小鼠原代肝细胞,检测FVP2对原代小鼠肝细胞活力的影响,对CCl4损伤的原代肝细胞ALT活力的影响。结果FVP2中、高剂量组血清AST活力值,肝匀浆液MDA值,高剂量组血清ALT活力较模型组有显著性差异(P<0.05);FVP2能显著增加小鼠原代肝细胞的活力(P<0.01);可使原代肝细胞培养液中的ALT活性明显低于模型组(P<0.01)。结论FVP2可能是CFVP肝保护作用的主要有效成分。     

硫普罗宁对大鼠创伤性脑损伤后应激性肝损害的保护作用

目的:探讨硫普罗宁在大鼠创伤性脑损伤后应激性肝损害的保护作用.方法:采用Feeney's自由落体撞击法建立创伤性脑损伤后应激性肝损害模型.于致伤后不同时间点(1,3,6,12和24 h)腹腔注射硫普罗宁300 mg·kg-1,测定大鼠肝组织超氧化物歧化酶(SOD)、丙二醛(MDA),血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST),并观察肝脏组织病理改变.结果:硫普罗宁各时间点给药组的大鼠肝细胞病理学改变显著减轻,肝组织SOD,MDA和血清ALT,AST有所改善,早期给药,作用越显著.结论:硫普罗宁作为抗氧化剂可用于防治创伤性脑损伤引起的应激性肝损害,尤其是在颅脑损伤早期应用效果更好.     

血红素加氧酶1对实验性肝硬化内毒素血症大鼠肝功能的影响

目的 探讨血红素加氧酶1(HO-1)在实验性肝硬化内毒素血症大鼠中的作用.方法 80只大鼠均分为4组:对照组为正常大鼠+脂多糖(LPS);其余3组用硫代乙酰胺(TAA)诱导大鼠肝硬化模型后,TAA组腹腔内注射生理盐水,TAA+LPS组注射LPS,HM组注射氯化高铁血红素+LPS.造模后检测AST和ALT.结果 与对照组比较,其余3组血浆中ALT和AST水平均明显升高(P＜0.05),且HM组＞TAA+ LPS组＞TAA组.结论 HO-1通过其氧化损伤和促纤维化,促进大鼠肝硬化内毒素血症时肝损害.     

纳洛酮预处理大鼠肝细胞对缺血再灌注损伤的影响

为探讨纳洛酮预处理大鼠后对其肝细胞缺血再灌注损伤的影响 ,短期对 Wistar大鼠腹腔内注射纳洛酮 ,然后制作缺血再灌注损伤模型 ,检测血清中谷丙转氨酶 (ALT)和谷草转氨酶 (AST)的含量 ,检测肝组织中丙二醛 (MDA)和超氧化物歧化酶(SOD)的含量 ,通过光镜、电镜观察 ,了解肝细胞形态学及超微结构的改变。结果显示 ,纳络酮预处理组血清中 AL T和 AST含量及肝组织中 MDA含量于缺血再灌注组 (P<0 .0 5 ) ,而肝组织中 SOD含量则高于缺血再灌注组 (P<0 .0 5 ) ,与假手术组无明显差异 ,肝细胞形态学异常改变也明显减轻     

前列地尔对D-氨基半乳糖致急性肝损伤大鼠肝匀浆IL-18

目的：观察前列地尔对D-氨基半乳糖致急性肝损伤大鼠肝匀浆IL-18水平升高的抑制作用。方法：32只SD大鼠一次性腹腔注射D-氨基半乳糖（D-Gal）1.4g&#8226;kg-1，建立急性肝损伤模型。造模后大鼠随机分为前列地尔注射液治疗组（Lipo PGE130μg&#8226;kg-1）和阴性对照组（灭菌生理氯化钠溶液），均为一次性腹腔注射。检测两组造模前和造模后6，12，24h血清谷丙转氨酶（ALT）、谷草转氨酶（AST）和肝匀浆IL-18的水平。结果：D-氨基半乳糖造模后，两组大鼠血清ALT，AST和肝匀浆IL-18值呈动态升高（P<0.05）。在造模后6，12和24h，前列地尔30μg&#8226;kg-1治疗组的血清ALT，AST和肝匀浆IL-18值均明显低于阴性对照组。结论：前列地尔对D-氨基半乳糖致急性肝损伤大鼠有肝脏保护作用，其机制可能与降低肝脏IL-18水平有关。     

氯化钆保护细菌脂多糖引发肝脏损伤及其可能机制研究

目的探讨氯化钆(Gadolinium chloride,Gd Cl3)在细菌脂多糖(Lipopolysaccharide,LPS)引起肝脏损伤的影响及其机制。方法将MT基因敲除小鼠(MT-/-)及与之对应的野生型小鼠(MT+/+)各分为4组:生理盐水对照组、LPS染毒组、Gd Cl3组和Gd Cl3预处理+LPS染毒组。动物腹腔注射LPS(10 mg/kg,腹腔注射)或生理盐水(NS,腹腔注射),此前24 h给予Gd Cl3(10 mg/kg,尾静肪注射)或NS预处理。注射LPS后24 h测定血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)活力以及一氧化氮(NO)含量,镉饱和法测定肝脏金属硫蛋白(MT)含量,进行肝组织病理学检查,判定不同处理对MT-/-小鼠及MT+/+小鼠肝脏影响的差异。结果注射LPS后,MT-/-小鼠及MT+/+小鼠血ALT、AST活力,血NO含量均较对照组升高,两种小鼠肝组织均出现大量炎性细胞浸润、肝细胞浊肿、空泡变性、灶性液化坏死、星状细胞增生。组织病理学检查结果显示MT-/-小鼠肝组织损伤程度较MT+/+小鼠更为严重。氯化钆、LPS均可以诱导MT+/+小鼠肝脏MT生成。Gd Cl3预处理可以同时降低LPS对MT-/-小鼠及MT+/+小鼠血ALT,AST活力,血NO含量的影响,可以减轻LPS对两种小鼠肝组织病理学损伤。结论 Gd Cl3预处理可以有效减轻由LPS引起的肝脏损伤,MT通过其对Kuffer细胞的抑制作用保护LPS引起的肝脏损伤。     

Protective effect of raloxifene on lipopolysaccharide and acid- induced acute lung injury in rats

Aim： To evaluate the protective effect of oral raloxifene on acute lung injury. Methods： Thirty adult, male Sprague-Dawley rats each weighing 180-210 g were used and divided into 3 groups： the raloxifene-lipopolysaccharide （LPS）-HCI group （n=10）, the LPS-raloxifene-HCl group （n=10）, and the placebo group （n=10）. All the rats were injected intraperitoneally （ip） with 5 mg/kg LPS, and raloxifene （30 mg/kg） was orally administered 1 h before and 14 h after LPS injection into the raloxifene-LPS-HCl and the LPS-raloxifene-HCl groups, respectively; the placebo group received nothing. Sixteen hours after LPS injection, all the animals were anesthetized and the femoral artery was cannulated. All the rats received a direct intratracheal （IT） injection ofHCl （pH 1.2; 0.5 mL/kg）. The mean arterial pressure （MAP） and blood gas concentrations were measured. Fifteen rats （5 in each group, respectively） underwent a micro positron emission tomography （microPET） scan of the thorax 4 h after HCI instillation. The wet/dry （W/D） weight ratio determination and histopathological examination were also performed. Results： The rats in the LPS-raloxifene-HCl group had a lower [^18F]fluorodeoxyglucose uptake compared with the rats in the placebo group （4.67±1.33 vs 9.01±1.58, respectively, P〈0.01）. The rats in the LPS-raloxifene-HCl group also had a lower histological lung injury score （8.20± 1.23 vs 12.6±0.97, respectively, P〈0.01） and W/D weight ratio （5.335±0.198 vs 5.886±0.257, respectively, P〈0.01） compared to the placebo group. The rats in this group also showed better pulmonary gas exchange and more stable mean arterial pressure （MAP） compared to the placebo group. Conclusion： Raloxifene provides a significant protective effect on acute lung injury in rats induced first by LPS ip injection and then by HCI IT instillation.     

二硫代氨基甲酸吡咯烷对小鼠急性凋亡性肝损伤的效应

目的探讨二硫代氨基甲酸吡咯烷(PDTC)对氨基半乳糖(GalN)/细菌脂多糖(LPS)诱导小鼠急性凋亡性肝损伤的作用及其分子机制。方法设置生理盐水(NS)组、GalN/LPS组、PDTC+GalN/LPS组和PDTC组。每组10只小鼠被用于观察LPS处理后72h内的动物死亡情况;每组6只小鼠经LPS处理后1.5h被取血、处死并留取肝脏,用RT-PCR检测肝脏组织TNF-αmRNA表达水平,用EMSA分析肝脏NF-κB结合活性,每组12只小鼠于LPS处理后8h取血、处死并留取肝脏,测定血清丙氨酸转氨酶(ALT)活力,用TUNEL技术检测肝脏细胞凋亡,并对肝组织切片行常规HE染色。结果GalN/LPS共处理升高小鼠血清ALT活力;肝脏组织病理学检查发现,GalN/LPS组小鼠肝脏严重充血、坏死并伴有大量炎性细胞浸润,肝脏组织TUNEL阳性细胞增多;在GalN/LPS共处理72h内有90%小鼠发生死亡,所有死亡小鼠均伴有肝脏严重充血。PDTC预处理抑制GalN/LPS诱导的肝脏NF-κB激活和TNF-α表达,但PDTC预处理反而加重GalN/LPS引起的小鼠肝脏细胞凋亡、进一步升高血清ALT活力、加重肝脏充血和坏死并加速小鼠死亡。结论NF-κB抑制剂PDTC通过抑制肝脏实质细胞NF-κB介导的抗凋亡机制加重GalN/LPS诱导的小鼠急性凋亡性肝损伤。     

β淀粉样蛋白对衰老模型大鼠学习记忆及大脑皮层内脂褐素含量的影响

为探讨 β淀粉样蛋白 (βAP ,于d 2 8双侧海马内各注射 4 μg)对D 半乳糖 (D Gal ,ip ,5 0mg·kg- 1·d- 1连续 4 2d)致衰老大鼠学习记忆行为的影响 ,检测了大鼠学习记忆行为 ,包括开场行为 ,Y 迷宫分辨学习和一次性被动回避反应 ,并用荧光光度计测定大脑皮层内脂褐素含量 .结果表明D Gal和βAP使大鼠在新异环境中自发活动和探究行为减少 ,学习记忆能力减退 ;同时大脑皮层内脂褐素含量增加 .βAP与D Gal联用使上述作用进一步增强 ,说明 βAP与D Gal有一定协同作用     

Amiodarone exposure during modest inflammation induces idiosyncrasy-like liver injury in rats: role of tumor necrosis factor-alpha

Amiodarone [2-butyl-3-(3',5'-diiodo-4'α-diethylaminoethoxybenzoyl)-benzofuran] (AMD), a class III antiarrhythmic drug, is known to cause idiosyncratic hepatotoxic reactions in human patients. One hypothesis for the etiology of idiosyncratic adverse drug reactions is that a concurrent inflammatory stress results in decreased threshold for drug toxicity. To explore this hypothesis in an animal model, male Sprague-Dawley rats were treated with nonhepatotoxic doses of AMD or its vehicle and with saline vehicle or lipopolysaccharide (LPS) to induce low-level inflammation. Elevated alanine aminotransferase (ALT), aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyltransferase activities as well as increased total bile acid concentrations in serum and midzonal hepatocellular necrosis were observed only in AMD/LPS-cotreated rats. The time interval between AMD and LPS administration was critical: AMD injected 16 h before LPS led to liver injury, whereas AMD injected 2-12 h before LPS failed to cause this response. The increase in ALT activity in AMD/LPS cotreatment showed a clear dose-response relationship with AMD as well as LPS. The metabolism and hepatic accumulation of AMD were not affected by LPS coexposure. Serum concentration of tumor necrosis factor-alpha (TNF) was significantly increased by LPS and was slightly prolonged by AMD. In Hepac1c7 cells, addition of TNF potentiated the cytotoxicity of both AMD and its primary metabolite, mono-N-desethylamiodarone. In vivo inhibition of TNF signaling by etanercept attenuated the AMD/LPS-induced liver injury in rats. In summary, AMD treatment during modest inflammation induced severe hepatotoxicity in rats, and TNF contributed to the induction of liver injury in this animal model of idiosyncratic AMD-induced liver injury.     

乙酰半胱氨酸对半乳糖胺致小鼠引起免疫性肝衰竭的治疗作用

目的 :观察乙酰半胱氨酸 (NAC)治疗小鼠免疫性肝衰竭的疗效。方法 :ICR小鼠随机分成 6组 :正常对照组、模型对照组、阳性对照组 ,乙酰半胱氨酸 2 5 ,5 0 ,10 0mg·kg- 1剂量组 ,每组 30只。除正常对照组外 ,每组腹腔注射半乳糖胺 (D galac tosamine ,Gal) 80 0mg·kg- 1,并同时腹腔注射脂多糖 (lipopolysacharides ,LPS) 10 μg·kg- 1,正常对照组、模型对照组、阳性对照组、给药组于注入Gal/LPS前、后 2h分别静脉注射生理盐水 ,生理盐水 ,促肝细胞生长素 (pHGF) 6 0mg·kg- 1,NAC 2 5 ,5 0 ,10 0mg·kg- 1。结果 :乙酰半胱氨酸能明显阻止Gal/LPS攻击 6h后引起小鼠血清转氨酶活力增高 ,改善凝血功能 ,使肝组织腐胺水平下降 ,肝组织损害减轻 ,降低小鼠死亡率 ,显示剂量依赖。结论 :乙酰半胱氨酸对Gal/LPS致小鼠免疫性肝衰竭有明显预防和治疗作用     

Bacterial lipopolysaccharide exposure augments aflatoxin B(1)-induced liver injury.

Bacterial endotoxin (lipopolysaccharide; LPS) given to animals in large doses results in pronounced, midzonal liver injury. Exposure to smaller, non-injurious doses of LPS augments the toxicity of certain hepatotoxicants. This study was conducted to delineate the development of injury in a rat model of augmentation of aflatoxin B(1) (AFB(1)) hepatotoxicity by LPS. At large doses (i.e., > 1 mg/kg, ip), AFB(1) administration resulted in pronounced injury to the periportal regions of the liver. Male, Sprague-Dawley rats (250-350 g) were treated with 1 mg AFB(1)/kg, ip or its vehicle (0.5% DMSO/saline) and 4 h later with either E. coli LPS (7.4 x 106 EU/kg, iv) or its saline vehicle. Liver injury was assessed 6, 12, 24, 48, 72, or 96 h after AFB(1) administration. Hepatic parenchymal cell injury was evaluated as increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in serum and from histologic examination of liver sections. Biliary tract alterations were evaluated as increased concentration of serum bile acids and activities of gamma-glutamyltransferase (GGT), alkaline phosphatase (ALP), and 5'-nucleotidase (5'-ND) in serum. At all times and for all markers, injury in rats treated with either AFB(1) or LPS alone was absent or modest. In the AFB(1)/LPS cotreated group, hepatic parenchymal cell injury was pronounced by 24 h and had returned to control values by 72 h. The injury began in the periportal region and spread midzonally with time. Furthermore, changes in serum markers indicative of biliary tract alterations were evident by 12 h and had returned to control values by 72 h. Thus, the nature of the hepatic lesions suggested that LPS potentiated the effects of AFB(1) on both parenchymal and bile duct epithelial cells.     

Systemic Effects of Inhaled Ultrafine Particles in Two Compromised,Aged Rat Strains

Epidemiological studies associate morbidity and mortality with exposure to particulate air pollution in elderly individuals with existing cardiopulmonary disease. These associations led to the hypothesis that inhaled particles can exert adverse effects outside of the lung, particularly on the cardiovascular system. We tested this hypothesis by examining the pulmonary and peripheral effects of inhaled ultrafine carbon particles in old rats that were injected with endotoxin (lipopolysaccharide, LPS) to model systemic gram-negative bacterial infection. Fischer 344 rats (23 mo) and spontaneously hypertensive (SH) rats (11–14 mo) were injected with LPS (2 mg/kg, ip) immediately before being exposed to inhaled ultrafine carbon particles for 6 h (150 μg/m³, CMD = 36 nm). Controls were injected with sterile saline or were sham exposed. Twenty-four hours after LPS injection, bronchoalveolar lavage (BAL) fluid, cells, and blood were obtained to assess endpoints of inflammation, oxidant stress, coagulability, and the acute-phase response. LPS did not cause an influx of neutrophils (PMNs) into the alveolar space, but did increase the number and percentage of circulating PMNs and the concentration of plasma fibrinogen in both rat strains. Inhaled ultrafine particles did not induce lung inflammation in either rat strain. In both strains, ultrafine particles (UFP) were found to decrease the number of blood PMNs, increase the intracellular oxidation of a fluorescent dye (DCFD) in blood PMNs, and affect plasma thrombin–anti-thrombin (TAT) complex and fibrinogen levels. UFP were also found to interact with ip LPS with respect to plasma TAT complex levels and blood PMN DCFD oxidation. Differences between the two rat strains were also found for TAT complex levels, BAL cell reactive oxygen species release, and DCFD oxidation in both BAL macrophages and blood PMNs. These results suggest that inhaled ultrafine carbon particles inhaled at concentrations mimicking high episodic increases in urban air can exert extrapulmonary effects in old rats and that they can change the systemic response to an inflammatory stimulus.     

Lipopolysaccharide augments aflatoxin B(1)-induced liver injury through neutrophil-dependent and -independent mechanisms.

Exposure to small, noninjurious doses of the inflammagen, bacterial endotoxin (lipopolysaccharide, LPS) augments the toxicity of certain hepatotoxicants including aflatoxin B(1) (AFB(1)). Mediators of inflammation, in particular neutrophils (PMNs), are responsible for tissue injury in a variety of animal models. This study was conducted to examine the role of PMNs in the pathogenesis of hepatic injury after AFB(1)/LPS cotreatment. Male, Sprague-Dawley rats (250-350 g) were treated with either 1 mg AFB(1)/kg, ip or its vehicle (0.5% DMSO/saline), and 4 h later with either E. coli LPS (7. 4 x 10(6) EU/kg, iv) or its saline vehicle. Over a course of 6 to 96 h after AFB(1) administration, rats were killed and livers were stained immunohistochemically for PMNs. LPS resulted in an increase in PMN accumulation in the liver that preceded the onset of liver injury. To assess if PMNs contributed to the pathogenesis, an anti-PMN antibody was administered to reduce PMN numbers in blood and liver, and injury was evaluated. Hepatic parenchymal cell injury was evaluated as increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in serum and from histologic examination of liver sections. Biliary tract alterations were evaluated as increased concentration of serum bile acids and activities of gamma-glutamyltransferase (GGT), alkaline phosphatase (ALP), and 5'-nucleotidase (5'-ND) in serum. Neutrophil depletion protected against hepatic parenchymal cell injury caused by AFB(1)/LPS cotreatment but not against markers of biliary tract injury. This suggests that LPS augments AFB(1) hepatotoxicity through two mechanisms: one of which is PMN-dependent, and another that is not.