High expressions of caveolins on the proliferating bile ductules in primary biliary cirrhosis

AIM: An increase in bile ductular structures is observed in diverse human liver diseases, especially in primary biliary cirrhosis (PBC). These structures harbor the progenitor cell component of the liver. Caveolins are cholesterol-binding proteins involved in the regulation of several intracellular processes including cholesterol transport. This study aims to examine the role of caveolin in PBC. METHODS: Immunohistochemical and Western blotting studies were performed on human liver specimens obtained from patients with PBC and normal liver samples. The expression of caveolin (CAV)-1 and -2 was determined using specific antibodies. RESULTS: In normal liver, scanty immunostaining for CAV 1 and -2 was observed in bile ductules. In PBC liver samples, the expression levels of CAV-1 and -2 were increased on proliferating bile ductules especially in stage 3 cases, but was sparse on interlobular bile duct in stage 1 specimens. Especially, the regenerating bile ductules at the interface of portal tracts and necrotic areas were immunostained intensely for CAV-1 and -2. These phenomena were confirmed by Western blot. CONCLUSION: The present results demonstrate increased expression of caveolins in proliferating bile ductules in PBC, which may be related to the homeostasis of cholesterol transport in regenerating bile ductules in PBC liver.     

原发性胆汁性肝硬化患者外周血单核巨噬细胞中Notch受体表达情况

目的探讨原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)患者外周血中巨噬细胞膜Notch受体表达情况及其与巨噬细胞极化的关系。方法随机抽取上海中医药大学附属曙光医院收治的确诊为PBC患者30例和健康体检者15名,分为早期PBC组、晚期PBC组和健康对照组,采集空腹血,流式细胞术法检测外周血M1和M2型巨噬细胞及膜表面Notch受体表达水平。结果早期PBC患者巨噬细胞膜表面Notch1受体表达较对照组高(P<0.05),而Notch2受体和Notch3受体表达水平与健康对照组比较,差异无统计学意义(P>0.05);晚期PBC患者巨噬细胞膜表面Notch1和Notch3受体表达较对照组高(P<0.05),但Notch2受体表达差异无统计学意义(P>0.05)。晚期PBC患者Notch1受体和Notch3受体表达水平较早期PBC患者高(P<0.05)。巨噬细胞极化M2/M1比值与Notch受体表达水平呈线性相关。结论PBC患者外周血巨噬细胞膜表面Notch受体表达异常,且与巨噬细胞极化呈直线相关。     

PBC患者granulysin基因和蛋白的表达及临床意义

目的:探讨颗粒溶素(granulysin,GNLY)在原发性胆汁性肝硬化和患者(PBC)中的表达及其意义.方法:采用TaqMan探针技术,以18SrRNA为内参照,测定60例PBC患者外周血中GNLYmRNA的含量,同时应用酶联免疫吸附法(ELISA),检测PBC患者血清中GNLY蛋白的水平,并以健康体检组(n=100)和乙型肝炎肝硬化组(n=60)为对照.结果:PBC组GNLYmRNA的平均拷贝数显著高于健康对照组[(2.7±2.5)×108vs(3.0±1.9)×107,P<0.01]和乙型肝炎后肝硬化组[(2.7±2.5)×108vs(4.7±3.6)×105,P<0.001).PBC患者血清中的GNLY蛋白表达显著高于健康对照组(15.48±3.24μg/Lvs4.76±2.32μg/L,P<0.01)和乙型肝炎后肝硬化组(15.48±3.24μg/Lvs2.57±1.84μg/L,P<0.01).结论:GNLY的表达与PBC的发生、发展存在一定的相关性,可用于PBC临床诊断.     

原发性胆汁性肝硬化中医证候分型文献分析

目的了解现有原发性胆汁性肝硬化临床研究文献中中医证候规律的特点。方法检索中国期刊全文数据库、万方数据库和维普数据库,收集并阅读全文,筛选出符合纳入标准的文献,提取证候类型和辨证依据等资料,进行描述性统计学分析。结果在检索出的78篇文献中,符合纳入条件的文献7篇,所涉及原发性胆汁性肝硬化中医证型共20种,排名在前6位的中医证型分别为：肝郁脾虚（34.85%）、肝肾阴虚（27.82%）、湿热瘀血（6.69%）、湿热蕴结（4.23%）、脾胃气虚（3.87%）和湿滞血瘀（3.52%）;证候分型要素共14个,排名前6位的分别为脾虚（40.49%）、肝郁（39.43%）、肝阴虚（31.34%）、肾阴虚（31.34%）、瘀血（25.70%）和湿热（17.25%）。结论中医研究原发性胆汁性肝硬化的论文水平较低,分型依据缺乏科学性。     

Differences in the efficacy of ursodeoxycholic acid and bile acid metabolism between viral liver diseases and primary biliary cirrhosis

AIM AND METHODS: The effects of ursodeoxycholic acid (UDCA, 600 mg/day) on liver function test values, and serum and urinary bile acids levels in hepatitis C virus-related chronic hepatitis (CH, n = 39) and liver cirrhosis (LC, n = 25), and in primary biliary cirrhosis (PBC, n = 25) were compared. RESULTS: The percentages of improvement in alanine transaminase (ALT) and gamma-glutamyl transpeptidase (gamma-GTP) in CH were almost the same in LC. The rates of improvement in ALT in PBC were negatively correlated with histological stages in the liver. Total serum bile acid levels in LC rose to the same extent as in CH, but the increases in PBC were significantly smaller at stages 3-4 than stages 1-2. The urinary levels of hydroxylated metabolites of UDCA only slightly increased in LC, but they increased significantly at PBC stages 3-4. CONCLUSIONS: The efficacy of UDCA was preserved in LC, but diminished at PBC stages 3-4. The poor enrichment of UDCA in the bile acid pool and extensive biotransformation of UDCA may cause the limited efficacy of UDCA in the cirrhotic stage of PBC.     

Case Report: Multiple Cancers: Hepatocellular carcinoma and adenocarcinomas of the common bile duct and the gall-bladder in a woman with primary biliary cirrhosis

We present a case of primary biliary cirrhosis associated with simultaneous triple cancers: a hepatocellular carcinoma and adenocarcinomas of the common bile duct and gall-bladder. A 70 year old Japanese woman, who had been diagnosed with primary biliary cirrhosis (stage 2 by Scheuer) 15 years before, was admitted to Koseiren Kamo Hospital in a comatose state. Laboratory data were as follows: the ammonia level was high (164.0 μg/dL), the antimitochondrial antibody showed a 320-fold increase, a high level of alpha-fetoprotein was indicated (2677 ng/mL), hepatitis B surface antigen was negative and hepatitis C antibody by enzyme immunoassay was negative, although a test for the RNA of hepatitis C virus by polymerase chain reaction was positive (10^3.5 copies/50 μL). The patient's condition gradually worsened and the patient died of liver failure. Autopsy showed triple cancers in the liver (hepatocellular carcinoma; trabecular type, moderately differentiated), the common bile duct (well-differentiated papillary adenocarcinoma) and the gall-bladder (well-differentiated papillary adenocarcinoma) with primary biliary cirrhosis (stage 4). Primary biliary cirrhosis has been believed to be a low risk for the development of hepatocellular carcinoma, despite the high risk of extrahepatic malignancy. The simultaneous occurrence of triple cancers with primary biliary cirrhosis, to the best of our knowledge, has never been reported. The present case may provide additional evidence for a predisposition to malignancy in primary biliary cirrhosis.     

Antibodies to E1 and E2/Protein X components of pyruvate dehydrogenase complex in sera of patients with primary biliary cirrhosis

Aims/Methods: Using purified E1 component of pyruvate dehydrogenase complex (PDC) from bovine heart, we measured the levels of anti-E1 anti-bodies in PBC sera using ELISA and determined the degree of inhibition that these antibodies exerted on E1 enzyme activity. We also estimated levels of anti-E2/Protein X (Pro-X) antibodies in PBC sera using purified E2 and Pro-X of PDC which were copurified with E1.Results/Conclusions: Anti-E1 antibodies were detected in 87.5% (35/40) of PBC sera. Some of these sera inhibited E1 enzyme antivity but inhibition did not correlate with levels of anti-E1 antibodies.A high positive correlation (r=0.918) was found between levels of anti-E1 and anti-E2/Pro-X antibodies, suggesting that anti-PDC antibody production was stimulated by PDC itself. Levels of IgG class anti-E2/Pro-X antibodies were significantly higher in sera of symptomatic PBC patients than in those of asymtomatic PBC patients. It was also found that patients who were positive for only IgM class anti-E2/Pro-X antibodies had early-stage PBC.     

熊去氧胆酸治疗原发性胆汁性胆管炎患者疗效及对血清Nrf2和HO-1水平的影响

目的 探讨熊去氧胆酸(UDCA)治疗原发性胆汁性胆管炎和肝硬化(PBC)患者疗效及对血清核因子相关因子2(Nrf2)和血红素加氧酶-1(HO-1)水平的影响.方法 2017年1月～2020年1月我院诊治的原发性胆汁性胆管炎患者31例和原发性胆汁性肝硬化患者34例(Child A级15例,Child B级12例,Chil...     

原发性胆汁性肝硬化患者肝组织转化生长因子β-1表达水平变化

目的 探讨原发性胆汁性肝硬化（PBC）患者肝组织转化生长因子-β1（TGF-β1）表达的变化。 方法 52例PBC患者接受超声引导下肝穿刺活检,选择病理科留取的正常肝组织20份作为对照,另选择20例健康人采集静脉血。采用免疫组化法检测肝组织TGFβ-1表达,采用ELISA法检测血清肿瘤坏死因子α（TNF-α）和白介素6（IL-6）水平。 结果 正常肝组织不表达或仅有少量TGF-β1表达,而PBC患者肝实质细胞胞浆内呈TGF-β1高表达;PBC患者血清TNF-α和IL-6水平分别为（28.71±13.54） pg/ml和（21.3±9.4） pg/ml,显著高于健康人【分别为（21.3±15.4） pg/ml和（2.1±1.6） pg/ml,P<0.01】;7例PBC肝组织肝纤维化S0期患者肝组织TGF-β1表达及血清TNF-α和IL-6水平分别为（0.5±0.2）10-2、（7.1±4.1） pg/ml和（5.1±1.0） pg/ml,显著低于12例S1期【（4.2±1.3） 10-2、（18.6±6.2） pg/ml、（11.5±3.6） pg/ml,P<0.01】、18例S2期【（6.9±1.2） 10-2、（27.3±9.9） pg/ml、（19.4±4.1） pg/ml,P<0.01】、9例S3期【（13.3±15.1） 10-2、（39.7±15.18） pg/ml、（27.3±8.1） pg/ml,P<0.01】和6例S4期【（21.2±17.1） 10-2、（53.4±17.3） pg/ml、（47.8±11.0） pg/ml,P<0.01】;15例Child-Pugh A级患者肝组织TGF-β1表达及血清TNF-α和IL-6水平分别为（1.9±1.6） 10-2、（12.2±3.1） pg/ml和（7.3±2.5） pg/ml,显著低于25例Child-Pugh B级【（15.9±13.6） 10-2、（32.9±8.6） pg/ml、（21.8±6.3） pg/ml,P<0.01】或12例C级患者【（22.6±18.5） 10-2、（49.1±19.3） pg/ml、（45.5±12.7） pg/ml,P<0.01】。 结论 PBC患者肝组织TGFβ-1表达及血清TNF-α和IL-6水平显著升高,可能与肝组织纤维化增生和/或肝功能受损有关。     

原发性胆汁性肝硬化患者外周血单个核细胞Toll样受体表达与调节性T细胞相关性分析

目的 探讨原发性胆汁性肝硬化(PBC)患者外周血单个核细胞(PBMC)Toll样受体2、4、9(TLR2,TLR4,TLR9)表达及PBMC中CD4+ CD25+调节性T细胞(Tregs)特点和影响因素.方法 采用流式细胞术检测PBC患者(52例)和健康对照者(22例)外周血PBMC中TLR2、4、9阳性细胞及Tregs比例,比较其在PBC患者和健康对照者中的差别并分析TLR与Tregs相关性.结果 PBC患者Tregs比例低于健康对照者[(1.53+1.33)vs(4.42±1.43),P＜0.0001],TLR4阳性细胞比例高于健康对照者[(36.95±3.53)vs(32.84±8.06),P=0.003];PBC患者Tregs比例与TLR9阳性细胞比例呈负相关(R2 =0.115,P=0.016),健康对照者Tregs比例与TLR2,4和TLR2,4,9联合表达阳性细胞比例呈负相关(R2=0.326,P=0.007;R2 =0.226,P=0.034).结论 PBC患者外周血PBMC中Tregs比例降低,TLR4表达升高,可能受肝硬化失代偿期腹腔感染的影响.     

Protein expression of double‐stranded RNA‐activated protein kinase (PKR) in intrahepatic bile ducts in normal adult livers,fetal livers,primary biliary cirrhosis,hepatolithiasis and intrahepatic cholangiocarcinoma

Abstract: Background/Aim: The protein expression of double‐stranded RNA‐activated protein kinase (PKR) in intrahepatic bile ducts has not been investigated. Methods: Immunohistochemistry and a semiquantitative scoring method in normal liver and biliary diseases were used for the investigation. Results: In “normal” adult livers (n=10), intrahepatic bile ducts were negative for PKR. In normal fetal livers (n=25), primitive biliary epithelia were almost negative for PKR. In primary biliary cirrhosis (PBC) (n=30), damaged bile ducts were frequently positive for PKR, while uninvolved bile ducts were negative. In hepatolithiasis (n=27), proliferated bile ducts were positive for PKR, and the PKR score correlated with the degree of proliferation. In cholangiocarcinoma (CC) (n=44), PKR expression was frequently noted, and the PKR score correlated with good differentiation of CC, being highest in well‐differentiated CC and lowest in poorly‐differentiated CC. The PKR score decreased in the following order: CC (mean PKR score=3.96), hepatolithiasis (2.56), PBC (1.60), normal fetal liver (0.40), and normal adult livers (0.00). The PKR expression in hepatocytes was “baseline” in normal adult livers, while moderately increased in fetal livers, PBC, hepatolithiasis and CC. Conclusions: Although the significance of these data is unclear, they suggest (i) that PKR is absent in bile ducts in normal adult and fetal livers, (ii) that PKR in bile duct cells newly emerges or increases in PBC, hepatolithiasis, and CC, (iii) that PKR accumulates in damaged bile ducts in PBC, (iv) that PKR increases in parallel with biliary cell proliferation in hepatolithiasis, and (v) that PKR expression correlates with differentiation in CC. PKR expression in intrahepatic bile ducts seems to be associated with inflammation or cell proliferation of the bile duct cells.     

柔肝健脾活血利胆法联合熊去氧胆酸治疗原发性胆汁性肝硬化疗效观察

[目的]观察柔肝健脾活血利胆法联合熊去氧胆酸（IH）CA）治疗原发性胆汁性肝硬化（PBC）的临床疗效及对生化指标的影响。[方法]33例PBC患者分为对照组16例和治疗组17例,分别予以UDCA和柔肝健脾活血利胆法联合UDCA治疗,观察临床疗效、分期和生化指标的关系及指标的改善情况。[结果]①2组治疗前后临床症状体征均有不同程度的改善,治疗组缓解症状较对照组更明显（P〈0．05,〈0．01）;②PBC早、中、晚期患者结果比较,总胆红素（TBil）渐进式上升,晚期患者丙氨酸氨基转移酶（ALT）显著下降,天冬氨酸转氨酶（AST）反而有所上升,ALT与TBil呈酶-胆分离现象,碱性磷酸酶（ALP）、γ-谷氨酰转移酶（GGT）在早期明显升高,晚期显著下降,血清清蛋白呈下降趋势;③治疗后2组患者生化指标均有所改善,而治疗组在改善血清TBil、GGT更加显著（P〈0．01）。[结论]柔肝健脾活血利胆法联合UDCA治疗PBC可以明显改善患者的临床症状、体征,提高治愈率及有效率,从而提高患者的生活质量,可望减少肝移植的机率。