硫代乙酰胺所致急性肝衰时内毒素血症与肝内胆汁淤积关系 …

本实验采用硫代乙酰胺（ｔｈｉｏａｅｔａｍｉｄｅ，ＴＡＡ）灌胃复制大鼠急性肝功能衰竭的动物模型，检测血中内毒素，转氨酶，脂蛋白－ｘ等含量以及胆汁流量，结果表明，大鼠发生了肝内胆汁淤积与内毒素血症（ＥＴＭ）并且两者呈正相关（Ｐ〈０．０５），同时肝细胞膜上Ｎａ＾＋－Ｋ＾＋－ＡＴＰ酶的活力明显降低，与ＥＴＭ含量呈负相关（Ｐ〈０．０５），结果提示：ＥＴＭ与肝内胆汁淤积的发生有密切关系。     

慢性胆道梗阻时肝脏病理变化的立体学研究

用立体学方法测得大鼠胆总管结扎后1W、2W、3W，其汇管区周围肝窦横径较对照组明显扩张（P＜0．01），增大幅度分别为67．34％、86．26％和94．81％。单位体积肝窦表面积较对照组明显减小（P＜0．01），减小幅度分别为31．07％、32．94％和36．21％。各结扎组中，肝细胞体积比较对照组肝细胞体积比减小（P＜0．05）；肝内纤维组织体积比较对照组肝内纤维组织体积比增加（P＜0．05）；肝内小血管体积比虽较对照组肝内小血管体积比增加，但无统计学差异（P＞0．05）。本实验证实：慢性肛道梗阻后肝内分流途径是汇管区周围扩张的肝窦。     

18月龄雌性大鼠下丘脑内源性阿片肽含量及其mRNA水平变化的研究

本文应用分子生物学和放射免疫测定方法，观察了老年前期雌性大鼠下丘脑、垂体和血浆中内阿片肽含量及下丘脑内阿片肽基因表达的变化．与5月龄青年大鼠比较，18月龄雌性大鼠下丘脑和血浆β-EP、L－ENK、Dy－nA含量减少，垂体β-EP、L－ENK含量升高，DynA含量降低（P＜0．05～0．01）．下丘脑POMC基因和proenkepbalin基因mRNA水平明显低于青年大鼠（P＜0．05）．结果提示，老年前期雌性大鼠下丘脑内阿片肽合成能力减退，衰老对下丘脑和垂体内阿片肽调节机制的影响有所不同．     

加味四逆散抗大鼠肝纤维化的实验研究

目的探讨加味四逆散抗大鼠肝纤维化的作用。方法采用二甲基亚硝胺（DMN）致肝纤维化的方法对健康雄性SD大鼠进行造模，以秋水仙碱为阳性对照组。经加味四逆散治疗4周后测血清丙氨酸转氨酶（ALT）、碱性磷酸酶（ALP）、谷氨酰转肽酶（GGT）、总蛋白（TP）、白蛋白（ALB）、透明质酸（HA）及Ⅳ型胶原（C—Ⅳ）水平，并检测肝组织羟脯氨酸（HyP），同时观察肝组织的病理改变。结果加味四逆散治疗后，可显著降低血清ALT（P〈0．01）、ALP（P〈0．01）、GGT（P〈0．05）、HA（P〈0．01）、C-Ⅳ（P〈0．05）及肝组织HyP（P〈0．01），升高TP（P〈0．05）及ALB（P〈0．01）。肝组织病理切片表明，用药后病理改变明显减轻。结论加味四逆散对实验性大鼠有明显的抗肝纤维化作用。     

内毒素诱发肝硬化大鼠发生肝性脑病的实验研究

目的：观察外源性内毒素诱发肝硬化大鼠发生肝性脑病的可能性及可能机制。方法：采用腹腔内小剂量内毒素一次性注射（３００μｇ／１００ｇＢＷ）诱发肝硬化大鼠发生肝性脑病。结果：小剂量内毒素腹腔内注射可以诱发肝硬化大鼠发生肝性脑病。内毒素注射后,动物一般行为状态及脑电图皆有明显变化,与其他肝性脑病模型或肝性脑病病人类似。同时,血氨和胰高血糖素水平显著升高。血浆内毒素含量与血氨、血胰高血糖素含量之间及血氨与务     

Caspase与妊娠期肝内胆汁淤积症的相关性研究

目的 探讨细胞凋亡因子Caspases（半胱天冬氨酸蛋白酶）增加是否与妊娠期肝内胆汁淤积症（ICP）的发病具有相关性。方法 采取剖宫产分娩的27例ICP孕妇（研究组）和30例正常孕妇（对照组）的胎盘组织。①用TUNEL原位标记术精确测定ICP孕妇（研究组）和正常孕妇（对照组）胎盘组织细胞凋亡指数。②用免疫组化法检测ICP孕妇（研究组）和正常孕妇（对照组）胎盘组织中Caspase-3，Caspase-9的表达情况。结果 研究组中合体滋养细胞中的凋亡指数高于对照组（P〈0．05），研究组促凋亡蛋白Caspase-3，Caspase-9在合体滋养细胞的阳性表达率高于对照组（P〈0．05）。结论 合体滋养细胞中的细胞凋亡增加与妊娠期肝内胆汁淤积症的发病具有相关性，可能在ICP的发病发展过程中具有一定作用。     

胆道梗阻对肝线粒体功能损伤机制的初步研究

目的：初步探讨胆道梗阻对肝线粒体功能损伤的机制。方法：复制犬胆道梗阻模型,观察梗阻的不同时段肝线粒体呼吸功能、钙含量、钙摄取率、丙二醛（MDA）含量及超氧化物歧化酶（SOD）含量的变化。结果：胆道梗阻2周、3周、4周及5周各组（Ⅱ－V组）分别与假手术组（Ⅰ组）比较：①肝线粒体呼吸控制率、钙摄取率及SOD含量均明显下降（Ⅱ组P＜0.05,其余组P＜0．01）②线粒体钙含量均明显升高（Ⅱ组P＜0．05,其余组p＜0．01）③线粒体MDA含量明显升高（各组P＜0.01）。相关分析显示,线粒体钙含量、MDA含量与线粒体呼吸控制率变化之间均呈现显著的负相关（P＜0．01）。结论：胆道梗阻后肝线粒体呼吸功能受到明显损伤,钙超载及脂质过氧化反应是造成线粒体外能损害的可能机制。     

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。     

大鼠腹腔注射人血清白蛋白免疫性肝纤维化模型的实验研究

目的 通过腹腔注射人血清白蛋白建立免疫损伤性大鼠肝纤维化模型。方法 雄性Wistar大鼠，体重110—120g，皮下多点注射人血清白蛋白（共4次。分别间隔14、10、10d）。10d后，腹腔注射人血清白蛋白（每周2次，共8周，剂量从5mg逐渐增至20mg）。在攻击前、攻击后15、30、60d和停止攻击后30、60、90、120d，分别留取肝组织和血清标本，进行肝病理检查和肝羟脯氨酸生化测定，采用放射免疫法测定肝纤维化血清指标透明质酸（HA）和层粘连蛋白（LN）含量。结果 攻击后。肝纤维化分级、肝羟脯氨酸含量、血清HA、LN均升高（P〈0．05）。随造模时间延长，肝纤维化分级逐渐加重（P〈0．05）、肝羟脯氨酸含量、血清HA逐渐升高（P〈0．01）。停止造模后，肝羟脯氨酸含量、血清HA明显降低（P〈0．01），但仍显著高于正常对照组（P〈0．01）。纤维化形成率为100％，纤维化持续时间120d以上。结论 此造模方法简单，肝纤维化成功率高，维持时间长，可用于抗纤维化药物的研究。     

肝炎患者血清透明质酸、Ⅲ型前胶原氨基端尾肽、层粘蛋白与Ⅳ型胶原的检测及其临床意义

分析了166例肝病患者血清透明质酸（HA）、Ⅲ型前胶原氨基端尾肽（PⅢNP）、层粘蛋白（Ln）与Ⅳ型胶原（Ⅳ－C）。结果是：HA、PⅢNP、Ln在各型肝病人中血清含量均高于正常组（P＜0．01），IV－C在除急性肝炎组外的其余各组血清含量均高于正常组（P＜0．05）。而HA、PⅢNP、Ⅳ－C的含量，慢活肝、肝硬化组的病人高于慢迁肝组的病人（P＜0．01）。肝硬化组病人Ln高于慢迁肝组病人（P＜0．05）。HA、PⅢNP、Ⅳ-C、Ln与SB、ALB有相关性，而与ALT无关，慢活肝组病人PⅢNP平均值高于肝硬化组的病人。提示这些肝硬化指标在反映肝纤维化程度上有实用意义，似乎PⅢNP更能功态反映肝纤维化的程度。但这些肝纤维化指标在不同程度上受肝细胞损害的影响。     

妊高征患者血浆ET水平与红细胞膜ATP酶活性的改变与发生肾病的关系

目的：探讨了妊高征肾病患者血浆内皮素（ET）水平和红细胞膜ATP酶活性的改变参与妊高征肾病的可能机制。方法：应用放射免疫分析和Reilni制膜法制定了32例妊高征肾病患者血浆ET水平和红细胞膜上Na^＋-K^＋-ATP酶和Ca^2＋-Mg^2＋-ATP酶活性的变化，并与70名妊高征无肾病组和35名正常孕妇作比较。结果：妊高征肾病组和无肾病组血浆ET水平非常显著地高于正常孕妇组（P〈0．01），而红细胞膜上Na^＋K^＋-ATP酶和Ca^2＋-Mg^2＋-ATP酶的活性均显著地低于正常孕妇组（P〈0．01），妊征肾病组与无肾病组有显著性差异（P〈0．05）。结论：妊高征肾病的发生与发展与血浆ET和红细胞膜上Na^＋-K^＋-ATP酶和Ca^2＋-Mg^2＋-ATP酶的活性有密切的关系。     

妊娠期妇女血清甘胆酸与血脂水平相关性研究

目的:探讨妊娠期妇女肝内胆汁淤积症(ICP)的发病率及甘胆酸(CG)与血脂、碱性磷酸酶(ALP)水平的相关性。方法:采用放射免疫分析检测甘胆酸,用酶法测定血脂,速率法测定ALP。结果:2040例妊娠期妇女中有18.5%CG值升高,其中有2.8%可确诊为ICP。ICP患者与正常妊娠组相比,TG、CHOL、LDL、ALP均明显升高,而HDL则略降低。结论:ICP为妊娠期常见并发症,应将CG作为常规产前检查项目。ICP患者结合分析其血脂、ALP水平,有助于病情判断。     

Detection of the cholestatic factor in the liver tissue of patients with acute intrahepatic cholestasis

A novel lymphokine, which we have designated as the cholestatic factor (CF). was produced from peripheral blood lymphocytes of patients with drug-induced allergic intrahepatic cholestasis by stimulation with a causative drug in the presence of the liver soluble fraction containing liver-specific lipoprotein (LSP). Marked reductions in bile flow and bile acid excretion were induced in rats by injecting CF through a mesenteric vein. In order to confirm the presence of CF in the liver tissue of patients, we attempted to detect this lymphokine by using the immunofluorescence technique. As a result, CF was found in the liver tissue of six out of 30 patients with acute intrahepatic cholestasis including one with hepatitis A type, one with hepatitis B type, one with hepatitis non-A non-B type and three with drug-induced allergic hepatitis. In contrast, CF was undetectable in the liver tissue of patients without intrahepatic cholestasis. These results may additionally support our assumption that CF plays an important role in the induction of intrahepatic cholestasis in various liver diseases.     

Increased microfilaments in hepatocytes and biliary ductular cells in cholestatic liver diseases

To assess the extent of microfilaments in cholestatic liver diseases we examined the cytoplasmic microfilaments in intrahepatic and extrahepatic cholestasis in man by electron microscopy. Study subjects were two patients with drug-induced intrahepatic cholestasis, three patients with intrahepatic cholestasis due to viral hepatitis, four patients with extrahepatic cholestasis due to stones of the common bile duct and two patients with primary biliary cirrhosis. Two biopsied specimens from patients without clinical or histological evidence of liver disease served as noncholestatic controls. The microfilaments in hepatocytes and biliary ductular cells were significantly increased in cholestasis compared with those in non-cholestatic controls. Well developed bundles of microfilaments were noted around the pericanalicular ectoplasm and seemed to be parallel to plasma membrane of the hepatocytes in cholestasis. In cholestasis, there were increased bundles of microfilaments around the periluminal region, lateral cell wall, and nucleus of biliary ductular cells. Two patterns of microfilaments bundles (fine microfilamentous network and spindle-shaped dense or clusters of microfilaments) were associated with cholestasis. The clustered form of microfilaments also seemed to be clearly associated with intracytoplasmic vacuoles containing bile salts. In conclusion, the increase of microfilaments in hepatocytes and biliary ductular cells may be the consequence of various forms of cholestasis. Further studies are needed to clarify the functional significance of increased microfilaments in cholestasis.     

Bioz.com在TURP患者血流动力学变化监测及TURS的早期诊治中的应用

目的 探讨Bioz.com在经尿道前列腺电切术（TURP）患者中早期诊治电切综合征（TURS）的应用价值。方法 选择我院择期行TURP的患者30例,行腰硬联合麻醉,记录患者麻醉前（T0）,麻醉后（T1）,灌洗液量3000 ml（T2）,灌洗液量6000 ml（T3）,灌洗液量9000 ml（T4）,灌洗液量12000 ml（T5）,灌洗液量15000 ml（T6）,灌洗液量18000 ml（T7）时的心率（HR）、收缩压（SBP）、舒张压（DBP）、搏出量（SV）、体血管阻力（SVR）、胸液成分（TFC）等血流动力学指标以及动脉血Na+,K+,Hb,Hct浓度。记录TURS发生情况。结果 与T0时比较,T1~T7时:HR升高（P＜0.05）,其中T1时升高最明显;SBP与DBP降低（P＜0.05）,其中T1时降低最明显;T2~T7时:Na+,Hb,Hct逐渐降低（P＜0.05）,K+逐渐升高（P＜0.05）;SV逐渐升高（P＜0.05）,SVR逐渐降低（P＜0.05）,其中T1时变化最明显;T2~T7时,TFC逐渐升高（P＜0.05）;与T1时比较,T2~T7时:HR逐渐降低（P＜0.05）;SBP与DBP逐渐升高（P＜0.05）;Na+,Hb,Hct逐渐降低,K+逐渐升高（P＜0.05）。SV逐渐升高（P＜0.05）,SVR逐渐降低（P＜0.05）,TFC逐渐升高（P＜0.05）。直线相关分析示,SV与Na+呈负相关（r=-0.948,P＜0.01）,与K+呈正相关（r=0.969,P＜0.01）,与Hb呈负相关（r=-0.976,P＜0.01）,与Hct呈负相关（r=-0.973,P＜0.01）。TFC与Na+呈负相关（r=-0.980,P＜0.01）,与K+呈正相关（r=0.996,P＜0.01）,与Hb呈负相关（r=-0.995,P＜0.01）,与Hct呈负相关（r=-0.997,P＜0.01）。所有患者均未发生TURS。结论 应用Bioz.com持续监测血流动力学变化,有助于通过无创的方法来早期预防TURS的发生。     

Activity of lysosomal enzymes in the bile and serum of mice with intrahepatic cholestasis

Lysosomal enzyme activity in the bile and blood serum was compared in mice with experimental intrahepatic cholestasis induced by α-naphthyl isothiocyanate and Triton WR 1339. Triton WR 1339 increases the synthesis of cholesterol (fatty acid precursor) in liver cells. The development of intrahepatic cholestasis was confirmed by the increase in activities of alkaline phosphatase and γ-glutamyltransferase in blood serum. Administration of Triton WR 1339 in a dose of 100 mg/100 g was followed by a 10-fold increase in β-galactosidase activity (hepatocyte lysosomal enzyme) in the bile, but not in the serum of mice. β-Galactosidase activity significantly increased in the bile, but decreased in the serum of mice after treatment with α-naphthyl isothiocyanate in a dose of 200 mg/kg. Our results indicate that intrahepatic cholestasis is manifested in increased secretion of lysosomal glycosidases into the bile. Bile components can aggravate damage to liver cells by affecting the processes of hepatocyte apoptosis and necrosis. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 5, pp. 496–499, May, 2008     

Neonatal intrahepatic cholestasis caused by citrin deficiency in Korean infants

Citrin is a liver-type mitochondrial aspartate-glutamate carrier encoded by the SLC25A13 gene, and its deficiency causes adult-onset type II citrullinemia and neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD). Here, the authors investigated clinical findings in Korean infants with NICCD and performed mutation analysis on the SLC25A13 gene. Of 47 patients with neonatal cholestasis, three infants had multiple aminoacidemia (involving citrulline, methionine, and arginine) and galactosemia, and thus were diagnosed as having NICCD. Two of these three showed failure to thrive. The laboratory findings showed hypoproteinemia and hyperammonemia, and liver biopsies revealed micro-macrovesicular fatty liver and cholestasis. The three patients each harbored compound heterozygous 1,638-1,660 dup/ S225X mutation, compound heterozygous 851del4/S225X mutation, and heterozygous 1,638-1,660 dup mutation, respectively. With nutritional manipulation, liver functions were normalized and catch-up growth was achieved. NICCD should be considered in the differential diagnosis of cholestatic jaundice in Korean infants.     

Pericanalicular hepatocytic and bile ductular microfilaments in cholestasis in man.

The cytoplasmic microfilaments of hepatocytes in the pericanalicular area and of bile ductular cells in various types of human cholestasis were examined and compared with those seen in the noncholestatic human liver. An increase in the number of hepatocytic microfilaments with an increase in the apparent density of the filamentous network and in the thickness of bundles of filaments was evident in the vicinity of the bile canaliculi, in the pericanalicular ectoplasm, and in the surrounding cytoplasm in all types of cholestasis but more so in intrahepatic cholestasis. This increase in the number of microfilaments was also seen around undilated canaliculi. Microfilaments were also increased in number beneath the surface of the bile ductular cells facing the cytoplasm but not beneath the rest of the plasma membrane. More microfilaments also were found around ductular cell nuclei. No striking differences were noted between intrahepatic and extrahepatic cholestasis. These findings suggest that microfilaments play a role that is not established in human cholestasis.     

The coagulation system contributes to alphaVbeta6 integrin expression and liver fibrosis induced by cholestasis

Chronic injury to intrahepatic bile duct epithelial cells (BDECs) elicits expression of various mediators, including the αVβ6 integrin, promoting liver fibrosis. We tested the hypothesis that tissue factor (TF)-dependent thrombin generation and protease activated receptor-1 (PAR-1) activation contribute to liver fibrosis induced by cholestasis via induction of αVβ6 expression. To test this hypothesis, mice deficient in either TF or PAR-1 were fed a diet containing 0.025% α-naphthylisothiocyanate (ANIT), a BDEC-selective toxicant. In genetically modified mice with a 50% reduction in liver TF activity fed an ANIT diet, coagulation cascade activation and liver fibrosis were reduced. Similarly, liver fibrosis was significantly reduced in PAR-1(-/-) mice fed an ANIT diet. Hepatic integrin β6 mRNA induction, expression of αVβ6 protein by intrahepatic BDECs, and SMAD2 phosphorylation were reduced by TF deficiency and PAR-1 deficiency in mice fed the ANIT diet. Treatment with either an anti-αVβ6 blocking antibody or soluble transforming growth factor-β receptor type II reduced liver fibrosis in mice fed the ANIT diet. PAR-1 activation enhanced transforming growth factor-β1-induced integrin β6 mRNA expression in both transformed human BDECs and primary rat BDECs. Interestingly, TF and PAR-1 mRNA levels were increased in livers from patients with cholestatic liver disease. These results indicate that a TF-PAR-1 pathway contributes to liver fibrosis induced by chronic cholestasis by increasing expression of the αVβ6 integrin, an important regulator of transforming growth factor-β1 activation.     

Evidence for microfilament involvement in norethandrolone-induced intrahepatic cholestasis.

An experimental study of norethandrolone (NED)-induced intrahepatic cholestasis was made. NED was infused via a portal vein catheter into rat liver in vivo, and measurements were made of bile flow. Liver specimens were taken at intervals for light microscopy and for transmission and scanning electron microscopy. Bile-canalicular-rich membrane fractions were prepared. The effects of NED were also examined in isolated hepatocytes in suspension culture. NED infusion induced total cholestasis by 3 hours. Canalicular alterations commonly associated with cholestasis were found in in vivo infused liver and in isolated hepatocytes. Pericanalicular microfilament changes were also noted in both, with loss of filament structure and replacement by a granular zone. In isolated canalicular membrane fractions prepared from NED-treated animals, the normal investment of pericanalicular filaments was no longer present. Loss of the bile canalicular ruthenium red surface coat was also noted. In view of the identical findings in isolated hepatocytes and in in vivo liver, obstruction and mechanical factors can be excluded as possible causes. The results raise the possibility that the mechanism of NED-induced cholestasis may be related to disaggregation and/or detachment of microfilaments from the canalicular membranes.