Liver involvement in subjects with rheumatic disease

The liver is often overlooked as a target organ, with pathology either secondary to an underlying disease or due to the toxicity of therapies and the medical complications of extrahepatic diseases. It is thus important for the clinical rheumatologist to be aware of the diagnostic procedure to monitor liver injury. Indeed, systemic rheumatologic diseases may be associated with liver abnormalities secondary to the presence of a coexisting autoimmune liver disease (particularly primary biliary cirrhosis or autoimmune hepatitis), the direct involvement of the liver parenchyma, or the impact of medical treatments (particularly methotrexate) on the liver. In addition, the rheumatologist should be aware of the impact of immunosuppressive agents on underlying viral infections, particularly viral hepatitis. We review herein the data on the role of the liver in the clinical management of systemic rheumatic diseases.     

Interleukin-5 inhibition of biliary cell chloride currents and bile flow

Recent studies have detected significant elevations of interleukin (IL)-5 mRNA in the liver parenchyma of patients with both primary biliary cirrhosis and acute rejection after liver transplantation. In both of these disorders, intrahepatic biliary epithelial cells (BECs) are the targets of injury. We hypothesized that BECs may themselves express IL-5 receptors that may modulate key biliary functions. RNAs coding for IL-5alpha and -beta receptors were amplified by RT/PCR from a biliary cell line derived from a human cholangiocarcinoma (Mz-ChA-1) and verified by DNA sequencing. IL-5 receptor distribution was detected immunocytochemically on Mz-ChA-1 cells, immortalized murine BEC, bile duct-ligated rat liver, and isolated cholangiocytes. Patch-clamp studies on Mz-ChA-1 cells showed that IL-5 inhibits 5'-N-ethylcarboxamidoadenosine-stimulated chloride currents. Additional functional studies showed that IL-5 inhibits secretin-induced bile flow. We conclude that BECs express IL-5 receptors and that IL-5 modulates BEC chloride currents and fluid secretion. Since IL-5 has previously been associated with cholestatic liver disease, we speculate that IL-5 may contribute to liver injury through its effects on biliary secretion.     

The hypoalbuminaemia of ovine fascioliasis: the influence of protein intake on the albumin metabolism of infected and of pair-fed control sheep.

Studies using ¹²⁵I-albumin and ⁵¹Cr-labelled plasma proteins showed that the hypoalbuminaemia which developed in sheep during the migratory stage of Fasciola hepatica infections was brought about by a combination of reduced albumin synthesis and plasma volume expansion. It was suggested that these changes were a reflection of the attendant liver damage and possibly of preferential utilisation of amino acids for immunoglobulin production. During the biliary stage of the disease, when the animals developed even more marked hypoalbuminaemia, increased albumin degradation arising from excessive plasma leakage into the gut were the outstanding features. The severity of these changes was closely linked to the state of the albumin pools which in turn was related to such factors as the plane of nutrition, appetite and fluke burden of the host. More albumin was catabolised by sheep with low fluke burdens, and in animals with the same level of infection, greater rates of catabolism were associated with a high protein intake. Sheep which catabolised most albumin became the least hypoalbuminaemic and survived longest. These animals also synthesised most albumin. It was shown that by impairing albumin synthesis, inappetence was an important additional factor in the hypoalbuminaemia of heavy infections, particularly if superimposed on a low protein diet. Nevertheless, irrespective of the size of their adult fluke burden, chronically-infected sheep were able to synthesise more albumin than pair-fed controls.     

Immunological specificity and biological action of biliary glucagon-like substance in the rabbit

A glucagon-like substance named biliary IRG2000 whose molecular weight is approximately 2,000 was isolated by gel filtration from rabbit bile. This substance showed a strong crossreactivity as equivalent to 25.7 +/- 5.1 ng/ml of porcine glucagon in RIA with antiserum 30K specificity. Biliary IRG2000 brought about a significant increase and delayed the response of blood glucose level in coexistence with porcine glucagon, though it has no appreciable effect on the glucose level when administered singly to the mouse intraperitoneally. The response with the coexistence of these materials was far greater than when porcine glucagon was given alone. In Mortimore's type rat liver perfusion, a significant rise in glucose concentration in effluent was also observed when a mixture of biliary IRG2000 and porcine glucagon was perfused. The rate of 125I-glucagon degradation was found delayed in the presence of biliary IRG2000 when examined in the rat. Thus the increase and delayed response of glucose level in coexistence of porcine glucagon with biliary IRG2000 may be explained by a suppressive effect of glucagon degradation due to biliary IRG2000.     

Functional state of the liver during simulation of the hemodynamic effects of microgravity on the human body

Radioisotope studies of the choleresis function of the liver, ultrasonic studies of the liver and contractile function of the gallbladder and gastroduodenoscopy were carried out in eight subjects after a 24-h stay in a 12° antiorthostatic position (AOP) simulating the hemodynamic changes in the abdominal cavity caused by microgravity. The dynamically hindered venous blood outflow from the liver induced in the AOP model caused activation of choleresis on an empty stomach. This activation manifested itself as an increase in the central perfusion zone of the liver parenchyma, dilation of the biliary ductules, and contraction of the gall-bladder, as well as choleresis into the duodenum. Activation of choleresis in the liver took place against the background of a reduction of the area of radioactive marker distribution in the liver and a decrease in the hepatocyte metabolic activity and the concentration function of the biliary excretion system. The functional characteristics of the liver in the AOP model reflected the reaction caused by changes in its blood content due to the changes in the body position negative to the gravity vector. The mechanism of the changes includes the occurrence of a dynamic venous plethora in the liver; centralization of hepatic blood flow, and activation of choleresis activity against the background of peripheral blood flow depletion, as well as the reduction of metabolic activity of hepatocytes and the concentration function of the biliary excretion system.     

Pharmacokinetics of kanamycin during targeted delivery to the liver in erythrocyte ghosts in animals with experimental acute cholecystitis

Pharmacokinetics of kanamycin was studied after its targeted delivery to the liver in autological erythrocyte ghosts on 25 noninbred dogs with experimental acute cholecystitis in comparison to the routine intravenous administration of the antibiotic in solution. Kanamycin concentrations in the tissues of the liver, pancreas, spleen, kidneys and lungs as well as in bile and blood serum were determined by the agar diffusion method 24, 48 and 72 hours after the last administration. It was found that the targeted delivery of kanamycin in blood shadows made it possible to provide high concentrations of the antibiotic for prolonged periods in the liver and biliary ducts and to more efficiently arrest the clinical manifestations of acute cholecystitis as well as normalize the laboratory indices. The data showed that using blood shadows as a reliable system for targeted delivery of antibiotics to the liver was advisable in purulent inflammatory affections of the biliary ducts.     

Transcriptome atlas of eight liver cell types uncovers effects of histidine catabolites on rat liver regeneration

Eight liver cell types were isolated using the methods of Percoll density gradient centrifugation and immunomagnetic beads to explore effects of histidine catabolites on rat liver regeneration. Rat Genome 230 2.0 Array was used to detect the expression profiles of genes associated with metabolism of histidine and its catabolites for the above-mentioned eight liver cell types, and bioinformatic and systems biology approaches were employed to analyse the relationship between above genes and rat liver regeneration. The results showed that the urocanic acid (UA) was degraded from histidine in Kupffer cells, acts on Kupffer cells itself and dendritic cells to generate immune suppression by autocrine and paracrine modes. Hepatocytes, biliary epithelia cells, oval cells and dendritic cells can convert histidine to histamine, which can promote sinusoidal endothelial cells proliferation by GsM pathway, and promote the proliferation of hepatocytes and biliary epithelia cells by GqM pathway.     

Pathogenesis of biliary fibrosis

Chronic cholestatic liver diseases such as primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) are associated with active hepatic fibrogenesis, and, ultimately, to the development of cirrhosis. However, the precise relationship between cholestasis, in its broad meaning, and liver tissue fibrosis is still poorly defined. Fibrogenesis is currently viewed as a dynamic process that appears strictly related to the extent and duration of parenchymal injury. This relationship is clearly evident in the presence of reiterative hepatocellular necrosis due to viral infection or alcohol abuse. It appears that “pure” intralobular intrahepatic cholestasis secondary to biliary secretory failure of the hepatocyte, in absence of hepatocellular damage, lobular inflammation and bile duct damage and/or proliferation, is not associated with marked and/or progressive liver tissue fibrosis. In contrast, marked and progressive liver tissue fibrosis always follows liver diseases characterized by chronic inflammatory bile duct damage as seen in PBC and PSC or chronic mechanical obstruction of the biliary tree. Overall, the fibrogenic process in these clinical conditions appears to be related to a more complex interaction between immune/inflammatory mechanisms, cytokine networks and the derangement of the homeostasis between epithelial and mesenchymal cells. The elucidation of these mechanisms is indeed crucial for the identification of potential diagnostic and therapeutic targets.     

Influence of tauroursodeoxycholic and taurodeoxycholic acids on hepatic metabolism and biliary secretion of phosphatidylcholine in the isolated rat liver

Studies were carried out using an isolated rat liver system to define: the contribution of exogenous phosphatidylcholine (PC) to biliary phospholipid secretion; and its hepatic metabolism during perfusion of the livers with conjugated bile salts with different hydrophilic/hydrophobic properties. A tracer dose of sn-1-palmitoyl-sn-2-[14C]linoleoylPC was injected as a bolus into the recirculating liver perfusate, under constant infusion of 0.75 mumol/min of tauroursodeoxycholate or taurodeoxycholate. The effects on bile flow, biliary lipid secretion, 14C disappearance from the perfusate and its appearance in bile, as well as hepatic and biliary biotransformation were determined. With both the bile salts, about 40% of the [14C]PC was taken up by the liver from the perfusate over 100 min. During the same period less than 2% of the given radioactivity was secreted into bile. More than 95% of the 14C recovered in bile was located within the identical injected PC molecular species. The biliary secretion of labeled as well as unlabeled PC, however, was significantly higher in livers perfused with taurodeoxycholate than tauroursodeoxycholate, while the reverse was observed with respect to bile flow and total bile salt secretion. The exogenous PC underwent extensive hepatic metabolization which appeared to be influenced by the type of bile salt perfusing the liver. After 2 h perfusion, the liver radioactivity was found, in decreasing order, in PC, triacylglycerol, phosphatidylethanolamine and diacylglycerol. In addition, the specific activity of triacylglycerol was significantly higher in tauroursodeoxycholate than in taurodeoxycholate-perfused livers (P less than 0.025), while the reverse was true for the specific activity of hepatic PC (P less than 0.01). Because taurodeoxycholate and tauroursodeoxycholate showed opposite effects on both biliary lipid secretion and hepatic PC biotransformations, we conclude that the hepatic metabolism of glycerolipids is influenced by the physiochemical properties of bile salts.     

Expression of neural cell adhesion molecule in human liver development and in congenital and acquired liver diseases

In the liver, neural cell adhesion molecule (NCAM) is a marker of immature cells committed to the biliary lineage and is expressed by reactive bile ductules in human liver diseases. We investigated the possible role of NCAM in the development of intrahepatic bile ducts and aimed at determining whether immature biliary cells can contribute to the repair of damaged bile ducts in chronic liver diseases. Therefore, we performed immunohistochemistry for NCAM and bile duct cell markers cytokeratin 7 and cytokeratin 19 on frozen sections of 85 liver specimens taken from 14 fetuses, 10 donor livers, 18 patients with congenital liver diseases characterized by ductal plate malformations (DPMs), and 43 cirrhotic explant livers. Duplicated ductal plates and incorporating bile ducts during development showed a patchy immunoreactivity for NCAM, while DPMs were continuously positive for NCAM. Bile ducts showing complete or patchy immunoreactivity for NCAM were found in cirrhotic livers, with higher frequency in biliary than in posthepatitic cirrhosis. Our results suggest that NCAM may have a function in the development of the intrahepatic bile ducts and that NCAM-positive immature biliary cells can contribute to the repair of damaged bile ducts in chronic liver diseases.     

Hepatitis B virus biology.

Hepadnaviruses (hepatitis B viruses) cause transient and chronic infections of the liver. Transient infections run a course of several months, and chronic infections are often lifelong. Chronic infections can lead to liver failure with cirrhosis and hepatocellular carcinoma. The replication strategy of these viruses has been described in great detail, but virus-host interactions leading to acute and chronic disease are still poorly understood. Studies on how the virus evades the immune response to cause prolonged transient infections with high-titer viremia and lifelong infections with an ongoing inflammation of the liver are still at an early stage, and the role of the virus in liver cancer is still elusive. The state of knowledge in this very active field is therefore reviewed with an emphasis on past accomplishments as well as goals for the future.     

Hepatitis B Virus Biology

Hepadnaviruses (hepatitis B viruses) cause transient and chronic infections of the liver. Transient infections run a course of several months, and chronic infections are often lifelong. Chronic infections can lead to liver failure with cirrhosis and hepatocellular carcinoma. The replication strategy of these viruses has been described in great detail, but virus-host interactions leading to acute and chronic disease are still poorly understood. Studies on how the virus evades the immune response to cause prolonged transient infections with high-titer viremia and lifelong infections with an ongoing inflammation of the liver are still at an early stage, and the role of the virus in liver cancer is still elusive. The state of knowledge in this very active field is therefore reviewed with an emphasis on past accomplishments as well as goals for the future.     

Vitamin E attenuates liver injury induced by exposure to lead, mercury, cadmium and copper in albino mice

Water pollution is the contamination of water resources by harmful wastes or toxins. Both community and private sources of drinking water are susceptible to a myriad of chemical contaminants. Heavy metals pollution of surface water can create health risks. The present study was aimed to investigate the effect of vitamin E supplementation on male mice exposed to a mixture of some heavy metals (lead, mercury, cadmium and copper) in their drinking water for seven weeks. Significant increases of blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT) were detected in heavy metals-treated mice. Histopathologically, the liver sections from heavy metals-treated mice showed severe changes including disarrangement of hepatic strands, rupture in hepatocytes, advanced hepatocellular necrosis, dilation and congestion of blood vessels with hemorrhage, dense lymphocytic infiltration round the central vein and dark stained hepatocytic nuclei indicating cell pycnosis. Administration of vitamin E at a dose of 50 IU/kg body weight, five times weekly improved the observed biochemical and histopathological changes induced by these heavy metals intoxication. Hence, the results of this study suggest that vitamin E protects against these heavy metals-induced liver injury and the attenuating effect of vitamin E may be due to its antioxidant activity.     

胆道梗阻合并胆道感染的病原学特征和危险因素及与肝脏损伤的关系研究

目的:探讨胆道梗阻合并胆道感染的病原学特征和危险因素,并分析胆道感染与肝脏损伤的关系。方法:回顾性分析250例胆道梗阻患者的临床资料,分析胆道梗阻合并胆道感染的病原菌分布和主要病原菌的耐药性,分析胆道梗阻合并胆道感染的危险因素,比较各组的肝功能指标[天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、谷氨酰转移酶(GGT)、直接胆红素(DBIL)]和肝纤维化指标[层黏蛋白(LN)、透明质酸(HA)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C)]。结果:250例胆道梗阻中共有132例合并胆道感染,感染率为52.80%,共分离出病原菌150株,以革兰阴性菌和革兰阳性菌为主。粪肠球菌对红霉素的耐药率最高,屎肠球菌对林可霉素的耐药率最高,均对万古霉素的耐药率最低;大肠埃希菌和肺炎克雷伯菌均对氨苄西林的耐药率最高,对妥布霉素的耐药率最低。年龄≥60岁、有胆道手术史、肝功能Child-Pugh评分≥11分是胆道梗阻合并胆道感染的危险因素(P<0.05)。胆道感染组的AST、ALT、GGT、DBIL、LN、HA、PCⅢ、Ⅳ-C水平高于无胆道感染组和对照组,且无胆道感染组高于对照组(P<0.05)。结论:胆道梗阻患者胆道感染的发生率较高,并且胆道感染会进一步加重胆道梗阻患者的肝脏损伤,临床应根据其病原学特征和危险因素做好相应的防治工作。     

The role of conjugation reactions in enhancing biliary secretion of bile acids.

Shortening the five-carbon carboxylic acid side chain of cholic acid by one methylene group gave rise to a bile acid (norcholate) that was not a substrate for the bile acid-conjugating enzymes. The metabolism and biliary secretion of norcholate in intact liver was examined in the isolated perfused rat liver system. When rat livers were perfused with 14-20 microM solutions of norcholate for 10 min, norcholate was found in the unconjugated form in liver, venous effluent and bile. Neither tauronorcholate nor glyconorcholate was detectable by high-pressure liquid chromatography or fast-atom-bombardment mass spectrometry. The kinetics of hepatic uptake and biliary secretion of norcholate was compared with that for cholate, taurocholate and chemically synthesized tauronorcholate. The latter three bile acids were completely cleared from the perfusate and efficiently secreted into the bile. However, norcholate was incompletely extracted from the perfusate, and this was shown to be at least partially due to its relatively lower rate of hepatic uptake. Furthermore, the rate of norcholate secretion into bile was greatly reduced relative to the secretion of cholate or chemically synthesized tauronorcholate, even though the concentration of norcholate in the liver was comparatively high. These data demonstrate that the conjugation of bile acids greatly facilitates their secretion into bile.     

NCK-associated protein 1 like (nckap1l) minor splice variant regulates intrahepatic biliary network morphogenesis

Impaired formation of the intrahepatic biliary network leads to cholestatic liver diseases, which are frequently associated with autoimmune disorders. Using a chemical mutagenesis strategy in zebrafish combined with computational network analysis, we screened for novel genes involved in intrahepatic biliary network formation. We positionally cloned a mutation in the nckap1l gene, which encodes a cytoplasmic adaptor protein for the WAVE regulatory complex. The mutation is located in the last exon after the stop codon of the primary splice isoform, only disrupting a previously unannotated minor splice isoform, which indicates that the minor splice isoform is responsible for the intrahepatic biliary network phenotype. CRISPR/Cas9-mediated nckap1l deletion, which disrupts both the primary and minor isoforms, showed the same defects. In the liver of nckap1l mutant larvae, WAVE regulatory complex component proteins are degraded specifically in biliary epithelial cells, which line the intrahepatic biliary network, thus disrupting the actin organization of these cells. We further show that nckap1l genetically interacts with the Cdk5 pathway in biliary epithelial cells. These data together indicate that although nckap1l was previously considered to be a hematopoietic cell lineage-specific protein, its minor splice isoform acts in biliary epithelial cells to regulate intrahepatic biliary network formation.     

Cell-mediated Immune Responses in Chronic Liver Diseases

Studies of the cell-mediated response to liver antigens, using the leucocyte migration test, in 163 patients with various liver disorders showed that abnormal responses were almost confined to active chronic hepatitis (53% abnormal), primary biliary cirrhosis (64%), and cryptogenic cirrhosis (29%). The test was also abnormal in five out of seven patients with jaundice due to drug hypersensitivity and in one patient with acute infectious hepatitis at a time when mitochondrial antibodies were present in the serum. More of those with active chronic hepatitis on prednisone or azathioprine had normal tests than of those who were untreated, and in 8 out of 10 examined serially during therapy there was an accompanying improvement in leucocyte migration. Abnormal responses to salivary gland or kidney antigens were also found in nearly half of those with features of Sjögren''s syndrome or renal tubular acidosis as part of a multisystem involvement—this, though occurring in cryptogenic cirrhosis, was found with greater frequency in active chronic hepatitis and primary biliary cirrhosis. These cell-mediated immune responses, perhaps triggered by the initial damage to the liver from viral or other agents, may be responsible both for the perpetuation of the liver disease and, because of common surface antigens, for the damage to other organs.     

Metastatic small cell lung cancer causing biliary obstruction

We report a case with metastatic small cell lung cancer which first manifested with biliary obstruction due to metastasis. Prognosis of patients presenting with jaundice due to hepatic parenchyma involvement is thought to be poor. However, the patient was successfully treated with percutaneous transhepatic biliary drainage and combination chemotherapy with reduced dosage. We believe this to be the first such case report, despite the frequency of metastasis to the liver from small cell lung cancer.     

Lernaeocera branchialis: a potential pathogen to cod ranching

A study was carried out to determine the effect of Lernaeocera branchialis on Atlantic cod infected in the laboratory and in the field and also to ascertain its effect on cod-ranching. Sixty-four percent (308) of 481 cod acquired infections in the laboratory and 33% (159) of the infected fish died over a 4-yr period. About 74% of the deaths occurred within 4 mo of the infection. Monthly samples of cod collected adjacent to a cod-ranching operation showed an initial prevalence of 30% that subsequently decreased in the following 2 mo to 15%. Prevalence of the infection also decreased among the initial field sample of cod that were kept alive, from 30 to 17% during the same 2-mo period and to 9% after 8 mo and was associated with death caused by the parasite. Cod examined at intervals after infection showed evidence of reduced weight gain, lower liver somatic index, liver lipid, and blood values than controls. A field sample taken from the same area during the summer of the following year indicated a prevalence of 12%. This higher than usual prevalence (4-6%) was associated with retention of the intermediate host, Cyclopterus lumpus, that provided an additional source of infective stages. It is likely that increased parasitism could affect the success of the cod-ranching operation in view of the parasite's devastating effects on its hosts.     

A genetic screen in zebrafish identifies the mutants vps18, nf2 and foie gras as models of liver disease

Hepatomegaly is a sign of many liver disorders. To identify zebrafish mutants to serve as models for hepatic pathologies, we screened for hepatomegaly at day 5 of embryogenesis in 297 zebrafish lines bearing mutations in genes that are essential for embryonic development. Seven mutants were identified, and three have phenotypes resembling different liver diseases. Mutation of the class C vacuolar protein sorting gene vps18 results in hepatomegaly associated with large, vesicle-filled hepatocytes, which we attribute to the failure of endosomal-lysosomal trafficking. Additionally, these mutants develop defects in the bile canaliculi and have marked biliary paucity, suggesting that vps18 also functions to traffic vesicles to the hepatocyte apical membrane and may play a role in the development of the intrahepatic biliary tree. Similar findings have been reported for individuals with arthrogryposis-renal dysfunction-cholestasis (ARC) syndrome, which is due to mutation of another class C vps gene. A second mutant, resulting from disruption of the tumor suppressor gene nf2, develops extrahepatic choledochal cysts in the common bile duct, suggesting that this gene regulates division of biliary cells during development and that nf2 may play a role in the hyperplastic tendencies observed in biliary cells in individuals with choledochal cysts. The third mutant is in the novel gene foie gras, which develops large, lipid-filled hepatocytes, resembling those in individuals with fatty liver disease. These mutants illustrate the utility of zebrafish as a model for studying liver development and disease, and provide valuable tools for investigating the molecular pathogenesis of congenital biliary disorders and fatty liver disease.