Hepatic clearance of tissue‐type plasminogen activator and plasma kallikrein in experimental liver fibrosis

Abstract: We have previously shown that tissue‐type plasminogen activator (tPA) and rat plasma kallikrein (RPK) share a common, but not unique, pathway for liver clearance. Aim: To evaluate the hepatic clearance of both proteases in experimental liver fibrosis. Methods: The hepatic clearance of these proteases was studied in porcine serum‐induced liver fibrosis using the isolated and perfused rat liver model. To better interpret the results, we also studied four other experimental groups: the turpentine oil‐induced acute‐phase response (AP group), AP group followed by GdCl₃ administration (AP/Gd group), CCl₄‐induced cirrhosis (CCl₄ group) and normal group. Results: The tPA clearance decreased significantly by both fibrotic and cirrhotic rat livers whereas the RPK clearance was not altered by the fibrotic rat liver. The hepatic clearance of tPA was reduced in the AP and AP/Gd groups; on the other hand, RPK clearance was increased in the AP group and, interestingly, this effect was neutralized by concomitant GdCl₃ administration. Conclusions: We observed that tPA and RPK clearances were affected differently by fibrosis as well as by different stimuli of the acute‐phase response, despite the fact that they share a common hepatic clearance mechanism in normal livers, and they were equally affected in cirrhosis.     

大鼠酒精性肝纤维化肝非实质细胞蛋白质组学研究

目的本文研究酒精对肝非实质细胞蛋白质表达的影响,以探讨酒精性肝纤维化的发病机制。方法大鼠酒精灌胃导致其发生肝纤维化。采用James染色法检测大鼠肝脏的病理学变化,通过Percoll密度梯度离心富集肝非实质细胞,再通过二维凝胶电泳(2DE)分离非实质细胞的蛋白质,经考马斯亮蓝(G250)染色,采用液相色谱串联质谱鉴定差异表达的蛋白质,并对部分差异蛋白质采用实时定量RT-PCR和免疫印迹的方法进行验证。对于2DE胶上的蛋白质点,采用两样本t检验的方法,对于RT-PCR分析,采用Mann-Whitney U检验进行统计分析。结果建立了酒精性肝纤维化大鼠模型,通过Percoll密度梯度离心纯化的非实质细胞中淋巴细胞、Kupffer细胞和内皮细胞分别富集了1.5、3.2和3.7倍。采用二维凝胶电泳法检测到了800多个蛋白质点,检测到具有2倍以上的差异蛋白质有26个,采用LC-MS法鉴定了21个非冗余蛋白质,对其中7个蛋白质的RT-PCR分析发现:ANXA3、CES3、ATPA和NDUFV2的mRNA水平和蛋白质组研究结果一致。结论本研究鉴定了一批与酒精性肝纤维化相关的蛋白质,可能为了解酒精性肝纤维化发病机制提供一些新线索。     

Hepatocellular carcinoma in cirrhotic liver in Trieste

One hundred consecutive cases of hepatocellular carcinoma (HCC) in cirrhosis observed at autopsy were studied and their pathological aspects were compared with those reported in the literature. The results, which are representative of HCC epidemiology in a geographical area where cirrhosis is mostly due to alcohol abuse, show that similarities in the architectural pattern of HCC and weight of the liver exist between our material and samples with different aetiology and epidemiology. A relationship between the histological grade of HCC and its propensity to metastasize was demonstrated. The reported better prognosis of clear cells per se could not be confirmed, although clear cell HCC occurred exclusively in grade 2. It was also demonstrated that the relationship between grading and staging was strongly influenced by the association of HCC with cirrhosis, which is a fact that is usually overlooked by the common staging (and grading) methods.     

Endothelin-induced vasoconstriction in isolated perfused liver preparations from normal and cirrhotic rats

Isolated, perfused rat liver preparations (IPRL), obtained from rats with carbon tetrachlorideinduced cirrhosis and normal controls, were used to investigate responses to the vasoactive peptide endothelin-1 (ET-1). The mean perfusion resistance (R) of cirrhotic IPRL was significantly greater than that of controls (2.63 ± 0.24 vs 1.54 ± 0.14 mmHg/mL per min per g; P < 0.01). Both control and cirrhotic IPRL demonstrated a concentration-related increase in resistance (ΔR) in response to ET-1, with a minimum effective concentration of approximately 3 × 10⁻¹¹ mol/L. The EC₅₀ (-log of the 50% effective concentration) was not significantly different between cirrhotic and control IPRL (8.48 ± 0.19 and 8.79 ± 0.11, respectively); however, the maximum response to ET-1 was significantly greater in cirrhotic preparations (R: 10.4 ± 2.2 vs 4.4 ± 0.5 mmHg/mL per min per g, P < 0.01; DR, 7.8 ± 2.1 vs 2.8 ± 0.4 mmHg/mL per min per g, P < 0.01). Following maximal stimulation by ET-1, the mean portal-hepatic venous pressure gradient at a physiological flow rate of 1 mL/min per g was approximately 90% greater across cirrhotic IPRL than that across normal IPRL (11.2 ± 2.0 vs 5.9 ± 0.9 mmHg, respectively; P < 0.05). These results support the hypothesis that endogenously released ET-1 has a significant influence on the portal vascular resistance of cirrhotic liver in vivo and has an important role in the pathogenesis of portal hypertension.     

Treatment of cirrhotic rats with epidermal growth factor and insulin accelerates liver DNA synthesis after partial hepatectomy

Prevention of postoperative hepatic failure is important after hepatic resection. In patients with cirrhosis, impaired liver function and regenerative capacity after major hepatic resection are associated with increased morbidity and mortality. In this study, a combination of epidermal growth factor (EGF) and insulin were used as hepatotrophic factors in an attempt to stimulate DNA synthesis after 70% hepatectomy (HTX). Regenerative capacity was evaluated in normal and cirrhotic rat liver by measuring DNA synthesis in vivo. Micronodular liver cirrhosis was established by the simultaneous oral administration of CCl₄ and phenobarbital. Epidermal growth factor plus insulin was injected subcutaneously immediately after and 12 h after HTX or sham operation was performed. Rats were killed 24 h after the operation and liver regeneration was estimated by [³H]-thymidine incorporation into DNA as well as an autoradiographic nuclear labelling index. Hepatectomy increased [³H]-thymidine incorporation significantly in both normal and cirrhotic rats. In cirrhotic rats, [³H]-thymidine incorporation after HTX was significantly lower than in normal rats and administration of a combination of EGF and insulin after HTX enhanced [³H]-thymidine incorporation. In conclusion, DNA synthesis 24 h after HTX is decreased in cirrhotic rats compared with normal rats and EGF supplementation with insulin accelerates DNA synthesis in hepatectomized cirrhotic rats. The data suggest that administration of combinations of exogenous hepatotrophic factors may play a useful role in the treatment of cirrhotic patients undergoing major hepatic resection.     

Kidneys in chronic liver diseases

Acute kidney injury (AKI), defined as an abrupt increase in the serum creatinine level by at least 0.3 mg/dL, occurs in about 20% of patients hospitalized for decompensating liver cirrhosis. Patients with cirrhosis are susceptible to developing AKI because of the progressive vasodilatory state, reduced effective blood volume and stimulation of vasoconstrictor hormones. The most common causes of AKI in cirrhosis are pre-renal azotemia, hepatorenal syndrome and acute tubular necrosis. Differential diagnosis is based on analysis of circumstances of AKI development, natriuresis, urine osmolality, response to withdrawal of diuretics and volume repletion, and rarely on renal biopsy. Chronic glomerulonephritis and obstructive uropathy are rare causes of azotemia in cirrhotic patients. AKI is one of the last events in the natural history of chronic liver disease, therefore, such patients should have an expedited referral for liver transplantation. Hepatorenal syndrome (HRS) is initiated by progressive portal hypertension, and may be prematurely triggered by bacterial infections, nonbacterial systemic inflammatory reactions, excessive diuresis, gastrointestinal hemorrhage, diarrhea or nephrotoxic agents. Each type of renal disease has a specific treatment approach ranging from repletion of the vascular system to renal replacement therapy. The treatment of choice in type 1 hepatorenal syndrome is a combination of vasoconstrictor with albumin infusion, which is effective in about 50% of patients. The second-line treatment of HRS involves a transjugular intrahepatic portosystemic shunt, renal vasoprotection or systems of artificial liver support.     

慢性乙型肝炎患者肝脏硬度值下降趋势与肝纤维化逆转结局的关系

目的 分析抗病毒治疗中慢性乙型肝炎患者肝脏硬度值变化趋势与肝纤维化逆转结局的相关性。方法 基于恩替卡韦初治慢性乙型肝炎患者临床随访队列(2013年7月-2015年9月),检测基线和治疗中每半年HBV DNA水平、肝功能、肝脏硬度值。根据更为严格的标准将治疗前后Ishak评分变化分为3组:确定逆转组,Ishak评分下降≥2;不确定组,Ishak评分下降0~1;进展组,Ishak评分升高>1。多组间比较采用单因素方差分析。采用分段线性混合模型拟合不同肝纤维化逆转结局的慢性乙型肝炎患者肝脏硬度值下降趋势,斜率的显著性及不同组间斜率的比较采用t检验。结果 共纳入239例慢性乙型肝炎患者,恩替卡韦治疗1.5年后,18例患者(7.5%)达到肝纤维化组织学确定逆转、196例(82.0%)为不确定、25例(10.5%)为进展。开始抗病毒治疗半年内肝脏硬度值下降率:确定逆转组为-36.3%[95%可信区间(95%CI):-52.8%^-19.7%],不确定组为-23.7%(95%CI:-29.7%^-17.8%),进展组为-12.6%(95%CI:-31.0%~5.9%)。在确定逆转组与不确定组,肝脏硬度值在治疗后的前6个月比治疗6个月后的下降速度更快(P值均<0.05)。结论 抗病毒治疗半年内肝脏硬度值的下降速度与肝纤维化逆转结局相关性较强,治疗半年时肝脏硬度值下降超过30%者发生肝纤维化确定逆转的可能性更大。     

FibroScan诊断不同类型慢性肝病肝纤维化的效果比较

目的比较Fibro Scan诊断不同类型慢性肝病肝纤维化的异同性。方法收集2012年10月-2015年1月就诊于北京大学第一医院的516例慢性肝病患者的临床资料,按照病因分为慢性乙型肝炎(CHB)组(n=305)、慢性丙型肝炎(CHC)组(n=117)及原发性胆汁性肝硬化(PBC)组(n=94),记录所有患者血常规、肝功能、肝脏弹性(FS)值以及腹部B超检查结果。按照FS值7.3 k Pa、7.3 k Pa≤FS值15 k Pa、FS值≥15 k Pa将每组患者分为不同的FS等级。计量资料和计数资料多组间比较采用KruskalWallis H检验,两组间比较采用Mann-Whitney U检验;FS值与年龄、性别、BMI、血常规、肝功能、超声各指标的相关性采用多重线性回归方法进行分析。结果 CHB组患者FS值的独立影响因素为性别、血红蛋白、AST、白蛋白、ALP、TBil、BMI、门静脉内径、腹水(P值均0.05);CHC组患者FS值的独立影响因素为血小板、ALT、ALP、GGT、总胆汁酸、肝表面光滑度、脾静脉内径、腹水(P值均0.05);PBC组患者FS值的独立影响因素为TBil、总胆汁酸、腹水(P值均0.05)。CHB和CHC患者不同FS值等级组间绝大多数检测指标的差异均有统计学意义(P值均0.05);PBC组中FS值7.3 k Pa与7.3 k Pa≤FS值15 k Pa组间仅有ALT(Z=-2.121,P=0.034)、AST(Z=-3.027,P=0.002)、脾长度(Z=-2.496,P=0.013)差异有统计学意义;7.3 k Pa≤FS值15 k Pa与FS值≥15 k Pa组间血小板(Z=-2.289,P=0.022)、白蛋白(Z=-2.185,P=0.029)、TBil(Z=-2.642,P=0.008)、脾厚度(Z=-3.317,P=0.001)、脾长度(Z=-2.010,P=0.044)、脾静脉内径(Z=-2.296,P=0.022)差异有统计学意义。结论不同病因影响FS值的关键因素有所不同,总胆汁酸、TBil是PBC患者FS值的重要影响因素,而肝功能状态对CHB和CHC患者FS值的影响更为显著,提示Fibro Scan诊断肝纤维化的阈值设定要根据不同病因病种而异。     

FibroScan检测与慢性肝病临床诊断的相关性研究

目的探讨FibroScan在慢性HBV感染者肝纤维化诊断中的作用。方法选择我院364例进行FibroScan检测的慢性HBV感染者,获取FibroScan检测值（FS值）,分析其与肝功能、肾功能和凝血功能指标及超声诊断等级的相关性。结果临床诊断为慢性乙型肝炎170例,肝硬化194例。肝硬化组FS值显著高于慢性乙型肝炎组（P〈0．05）。将FS值与临床检验指标进行多重线性回归分析,Fs值与ALB、国际标准化比值、CHE、ALP、ALT之间具有相关性,其中ALB对Fs值影响最大。不同超声诊断等级分组的Fs值差异有统计学意义（P〈0．05）,FS值与超声诊断等级分组之间相关性较好（ra=0．670）。超声诊断为脾大但无肝硬化患者的FS值高于超声诊断无脾大、无肝硬化的患者（P〈0．05）。结论FibroScan在诊断慢性HBV感染者纤维化严重程度方面有较好的应用价值。     

乙型肝炎病毒相关慢加急性肝衰竭患者肝纤维化、肝功能、病毒学指标及甲胎蛋白水平与预后的关系

目的初步探讨乙型肝炎相关慢加急性肝衰竭(HBV-ACLF)患者血清肝纤维化、肝功能、病毒学指标及甲胎蛋白(AFP)水平与预后的关系。方法根据预后将185例HBV-ACLF患者分为好转组(116例)和无效组(69例),检测血清肝纤维化和肝功能指标、病毒学指标、AFP水平,分析肝纤维化等指标与预后的关系。结果好转组和无效组性别、HBV基因型、HBeAg阳性率、HBV DNA载量、ALT、Alb、CHE比较差异无统计学意义。好转组和无效组平均年龄分别为(43.3±10.1)岁和(48.7±10.1)岁,TBil分别为(295.9±99.6)μmol/L、(355.4±136.8)μmol/L,凝血酶原活动度(PTA)分别为(34.5±7.9)%、(30.4±7.6)%,AFP分别为85(9～4760)ng/L、26(4～529)ng/L,差异皆有统计学意义(P值分别为0.006、0.009、0.0007、0.000),肝纤维化指标中,透明质酸(HA)、Ⅲ型前胶原(PcⅢ)与Ⅳ型胶原(C-Ⅳ)好转组和无效组差异无统计学意义,层粘连蛋白(LN)好转组和无效组分别为657(45～1616)μg/L、306(29～1724)μg/L,差异有统计学意义(P<0.001)。结论血清LN、AFP、TBil、PTA水平、年龄可能对HBV感染慢加急性肝衰竭患者的预后判断有一定意义。     

CASE REPORT: Primary hepatic lymphoma associated with chronic liver disease

We report on a case of primary hepatic lymphoma that developed in a patient with chronic hepatitis C. Given that Japan is an area endemic for both hepatitis B and C viruses, we reviewed 51 Japanese cases of primary hepatic lymphoma, addressing the question as to whether the Japanese cases have unique characteristics and whether there is a causal relationship to the presence of chronic liver disease. Primary hepatic lymphoma most commonly affected middle-aged males. Presenting symptoms and physical findings were non-specific. Aminotransferases tended to stay in the low range compared with marked increases in lactate dehydrogenase. Sixteen patients (31%) had chronic liver disease, eight had liver cirrhosis and eight had chronic hepatitis, suggesting that there is a possible aetiological link between chronic liver disease and primary hepatic lymphoma.     

肝脏瞬时弹性值评估慢性乙型肝炎肝纤维化程度及肝脏储备功能的临床研究

目的探讨慢性乙型肝炎(CHB)患者FibroScan与肝组织纤维化面积之间的相关性,以评价FS值对肝纤维化程度测定的意义,分析乙型肝炎后肝硬化患者的FS值变化与肝脏储备功能评价系统CTP分级的关系。方法前瞻性研究正常对照组30例、肝病组113例(CHB患者62例及其所致肝硬化患者51例)FS值,对研究对象行腹部B超检查后,应用肝脏瞬时弹性超声进行FS值测定,收集肝硬化患者入院24 h内的临床资料,计算CTP分值,根据评分分为A、B、C 3级,同时采用计算机辅助数字图像分析法检测肝组织标本的纤维化程度,分析肝病组FS值与肝纤维化面积之间的相关性以及肝硬化患者FS值变化与CTP分级的关系。结果 (1)CHB患者组的FS值与正常对照组比较,肝硬化患者的FS值与CHB组比较,差异均有统计学意义(P<0.05);(2)在肝硬化组中FS值随着CTP分级升高而升高,ChildA、B、C 3组之间FS值差异均有统计学意义(P<0.05);肝病组的FS值与肝纤维化面积有显著的线性正相关性,且相关系数较高(r=0.804,P<0.01)。结论 FS弹性值与肝纤维化面积有很好的相关性,肝硬化患者的FS值随着CTP分级的上升而增加,FS值可以评估CHB肝纤维化程度,在一定程度上可以评估肝硬化患者的肝脏储备功能。     

The protective role of Hepatopoietin Cn on liver injury induced by carbon tetrachloride in rats*

Background: Hepatopoietin Cn (HPPCn) is a member of the leucine‐rich acidic nuclear protein family (LANP), and studies of partially hepatectomized (PH) mice show that levels of HPPCn mRNA increase following liver injury. Furthermore, the recombinant human protein (rhHPPCn) was shown to stimulate hepatic DNA synthesis and activate signaling pathways involved in hepatocyte proliferation in vitro and in vivo. Aim: The aim of the study was to evaluate the protective effect of rhHPPCn on liver injury and fibrosis induced by carbon tetrachloride (CCl4) injection. Methods: Wistar rats weighing 200 g were given a single and repeated intraperitoneal injections of CCl4. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity in rat serum were measured using biochemical assay. Hepatic hydroxyproline (Hyp) level was determined in the hydrolysates of liver samples. Immunostaining and Masson's trichrome staining were conducted to evaluate hepatocyte proliferation and fibrosis. Results: The results showed that exogenous rhHPPCn could alleviate hepatocyte necrosis and protect the liver from the development of fibrotic lesions by proliferation stimulation. Additionally, HPPCn could reduce ALT/AST levels in rat serum following single and repeated CCl4 injection. Conclusion: It was suggested that HPPCn could protect hepatocytes from injury induced by CCl4 as a proliferation stimulator.     

Potentiation of carbon tetrachloride-induced liver damage by dibutylyl-3',5'-cyclic AMP in unstarved rats

It has been reported that carbon tetrachloride-induced liver damage is potentiated by starvation partly due to fat accumulation in the liver and a decrease in hepatic reduced glutathione concentration and that dibutylyl-3′,5′-cyclic AMP (DBcAMP) affects fuel metabolism and decreases hepatic reduced glutathione. We investigated the effects of DBcAMP on carbon tetrachloride-induced liver damage both in unstarved and starved rats. In unstarved rats, intraperitoneal administration of DBcAMP potentiated an increase in serum alanine aminotransferase activity and fatty vacuolization in the liver, both of which were induced by carbon tetrachloride. Hepatic reduced glutathione concentration was also reduced by DBcAMP, although the change was not significant. In contrast, the administration of DBcAMP in starved rats did not affect carbon tetrachloride-induced changes in serum alanine aminotransferase activity, histological alterations and hepatic reduced glutathione concentration. Administration of DBcAMP to control rats induced different responses in unstarved control rats compared with starved control rats: in unstarved rats, blood glucose concentration decreased but serum free fatty acid concentration increased, whereas in starved rats, blood glucose concentration increased and serum free fatty acid concentration decreased. It was suggested that DBcAMP potentiated carbon tetrachloride-induced liver damage in unstarved rats, probably due to hepatic fat accumulation and a decreased hepatic reduced glutathione concentration. The former could increase the affinity of the liver for carbon tetrachloride and the latter could accelerate carbon tetrachloride-induced lipid peroxidation. It was also suggested that DBcAMP failed to affect carbon tetrachloride-induced liver damage in starved rats, probably because starvation had already decreased hepatic glutathione concentration and DBcAMP had different effects on fuel metabolism compared with effects observed in unstarved rats.     

银杏叶提取物对实验性大鼠肝纤维化的逆转作用

[目的]观察银杏叶提取物(EGb)对四氯化碳(CCl4)诱导大鼠肝纤维化病理过程的逆转作用.[方法]采用CCl4腹腔注射诱导大鼠肝纤维化模型成功后,予EGb或0.85%氯化钠灌胃,8周后用苏木精-伊红染色、苦味酸-酸性品红染色和网状纤维染色观察大鼠肝脏病理变化,酶动力法检测肝功能,SP免疫组化法检测肝组织中α-肌动蛋白(α-SMA)、转化生长因子β1(TGF-β2)、Ⅰ型胶原表达的变化,RT-PCR法检测肝组织中TGF-β1、Ⅰ型胶原、金属蛋白酶组织抑制因子(TIMP1)的表达情况.[结果]EGb可促进大鼠肝纤维化的逆转,使肝组织中α-SMA、TGF-β1、Ⅰ型胶原、TIMP1的表达降低(P＜0.01);肝功能改善(P＜0.01);肝组织纤维化程度分级好转,胶原纤维、网状纤维所占面积显著缩小(P＜0.01).[结论]EGb能有效逆转大鼠肝纤维化病理过程,其作用机制为抗脂质过氧化、保护肝细胞和抑制肝星状细胞活化,抑制TGF-β1、Ⅰ型胶原、TIMP1的合成,从而逆转纤维化的形成.     

Predictors of peri-opertative morbidity and liver dysfunction after hepatic resection in patients with chronic liver disease

BackgroundHepatic resection in patients with chronic liver disease (CLD) is associated with a risk of post-operative liver failure and higher morbidity than patients without liver disease. There is no universal risk stratification scheme for CLD patients undergoing resection.ObjectivesThe aim of the present study was to evaluate the association between routine pre-operative laboratory investigations, model for end-stage liver disease (MELD), indocyanine green retention at 15 min (ICG15) and post-operative outcomes in CLD patients undergoing liver resection.MethodsA retrospective review of patients undergoing resection for hepatocellular carcinoma (HCC) at the University Health Network was preformed. ICG15 results, pre- and post-operative laboratory results were obtained from clinical records. Adjusted odds ratios (AOR) were calculated for associations between pre-operative factors and post-operative outcomes using multivariate logistic regression adjusting for patient age and number of segments resected.ResultsBetween 2001 and 2005, 129 CLD patients underwent surgical resection for HCC. Procedures included 51 (40%) resections of ≤2 segments, 52 (40%) hemihepatectomies and 25 (19%) extended hepatic resections. Thirty- and 90-day post-operative mortality was 1.6% and 4.1%, respectively. Prolonged (>10 days) hospital length of stay (LOS) was independently associated with an ICG15 >15% {AOR [95% confidence interval (CI)]= 8.5 (1.4–51)} and an international normalized ratio (INR) > 1.2 [AOR (95% CI) = 5.0 (1.4–18.6)]. An ICG15 > 15% and MELD score were independent predictors of prolonged LOS. An ICG15 > 15% was also independently associated with MELD > 20 on post-operative day 3 [AOR (95% CI) = 24.3 (1.8–319)].ConclusionsElevated ICG retention was independently associated with post-operative liver dysfunction and morbidity. The utility of ICG in combination with other biochemical measures to predict outcomes after hepatic resection in CLD patients requires further prospective study.A possible role for ICG clearance in predicting outcome following hepatic resection for hepatocellular carcinoma     

他汀类药物在慢性肝病中的应用

对他汀类药物的肝脏安全性及其在慢性肝病中的应用进行评述,包括肝脏病专家对他汀类药物治疗血脂紊乱的肝脏安全性评估意见,以及肝脏转氨酶持续异常、非酒精性脂肪性肝病、慢性丙型肝炎、原发性胆汁性肝硬化、原发性肝细胞癌及肝移植患者使用他汀的效果和安全性.     

基于逍遥散及其拆方研究“肝病实脾法”对肝纤维化大鼠肠道菌群的影响

目的通过逍遥散及其拆方对肝纤维化大鼠肠道菌群结构、门静脉内毒素水平的变化,探讨"肝病实脾法"抗肝纤维化的可能作用机制。方法将健康Wistar大鼠70只随机分为空白组10只,模型组、实验组和对照组各20只,采用牛血清白蛋白尾静脉注射8周建立免疫性肝纤维化大鼠模型。实验组予逍遥散中药颗粒剂进行灌胃治疗,对照组予逍遥散去健脾类中药颗粒剂灌胃。观察各组血清转氨酶、肝组织病理学、门静脉内毒素及肠道菌群指纹图谱分析(ERIC-PCR)。满足方差齐性条件的计量资料多组间比较采用方差分析,进一步2组间比较采用LSD-t检验,方差不齐时用Tamhane's法。相关性检验采用Pearson相关分析。结果与空白组比较,模型组大鼠肠道菌群多样性及其构成发生改变,门静脉内毒素水平升高[(0.421±0.170)EU/ml vs(0.784±0.180)EU/ml],差异有统计学意义(P0.01),且门静脉内毒素与肝组织胶原纤维面积百分比呈正相关(r=0.736,P0.01);与模型组比较,实验组ALT、AST、门静脉内毒素水平有所下降[(73.25±10.90)U/L vs(59.84±9.60)U/L,(135.36±31.41)U/L vs(107.43±17.71)U/L,(0.784±0.180)EU/ml vs(0.576±0.220)EU/ml],差异均有统计学意义(P0.01或P0.05),且肠道菌群多样性及构成与正常大鼠相似;对照组与模型组、实验组在ALT、AST、门静脉内毒素水平方面比较,差异均无统计学意义(P值均0.05)。结论肝纤维化大鼠出现肠道菌群失调,并引起门静脉内毒素的升高,是慢性肝损伤病理过程中"肝病传脾"的发生机制之一。而逍遥散能够改善肝纤维化,部分恢复肠道菌群正常结构,降低内毒素;而去掉健脾药物将减弱逍遥散对肝脏的保护作用,说明恢复肠道菌群,降低门静脉内毒素可能是"肝病实脾法"抗肝纤维化的重要作用机制之一。     

Duplex-Doppler assessment of cirrhosis in patients with chronic compensated liver disease

Portal venous flow velocity (PFV) was measured with duplex-Doppler equipment in 50 normal subjects and in 117 patients with suspected chronic liver disease who showed no evidence of decompensation such as ascites, hepatic encephalopathy, jaundice or oesophageal bleeding. All the patients underwent percutaneous liver biopsy which demonstrated non-cirrhotic liver disease in 58 cases (CH-patients: steatosis 8, persistent chronic hepatitis 8, active chronic hepatitis 42) and liver cirrhosis in the other 59 cases (LC-patients). The normal subjects and the CH-patients had similar values of max-PFV and mean-PFV (max-PFV 26.7±3.2 and 25.7±3.4 cm/s respectively; mean-PFV 22.9±2.8 and 22.4±3.8 cm/s respectively). The LC-patients’ values (max-PFV 19.3±3.5; mean-PFV 16.9±2.9) were significantly lower than those of the normal subjects (P<0.001) and of the CH-patients (P<0.001). Considering the normal max-PFV to be in the range 20–33.1 cm/s (mean±2 s.d. of the normal subjects, 95% confidence limits), max-PFV was reduced in 0/50 normal subjects, 1/58 CH-patients and 39/59 LC-patients (66.1% sensitivity; 98.2% specificity). In conclusion, the duplex-Doppler measurement of PFV is of great interest in the diagnostic study of patients with suspected chronic compensated liver disease and in the early diagnosis of cirrhosis. A low max-PFV is a reliable pointer to liver cirrhosis, whereas a normal max-PFV indicates a non-cirrhotic liver disease but is less probative. Each centre should standardize normal PFV values in order to establish their own threshold value for diagnosing liver cirrhosis.     

复方黄根对四氯化碳所致大鼠慢性肝损伤的保护作用

[目的]研究复方黄根对大鼠慢性肝损伤的保护作用及可能机制.[方法]制备大鼠四氯化碳(CCl4)慢性肝损伤模型,观察复方黄根对肝损伤大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸转氨酶(AST)活性,总蛋白(TP)、清蛋白(Alb)和羟脯氨酸(Hyp)水平的变化以及肝组织匀浆超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)水平的影响,放免法检测透明质酸(HA)、Ⅲ型前胶原肽(PⅢP)水平,免疫组化法测肝组织转化生长因子-β1(TGF-β1)的表达,并观察肝组织病理学改变.[结果]复方黄根可显著降低CCl4所致大鼠慢性肝损伤血清中ALT、AST、HA、PⅢP、Hyp水平,升高血清中Alb、TP水平;升高肝组织中SOD、GSH-Px的活性,并可降低MDA的水平;免疫组化结果表明复方黄根能抑制TGF-β1表达;病理观察结果能减轻慢性肝损伤的肝脏损伤程度.[结论]复方黄根有明显的保肝和抗肝纤维化作用,其作用机制可能与抗脂质过氧化和抑制肝组织TGF-β1的表达有关.