血清胆汁酸谱系测定在肝硬化诊断中的应用价值

  目的  建立高效液相色谱 – 串联质谱法快速检测血浆中6种初级胆汁酸的含量。  方法  血浆经甲醇沉淀蛋白后,用Thermo Accucore C₁₈色谱柱（100 mm × 4.6 mm,2.6 μm）进行色谱分离,流动相为甲醇 – 水（0.1 %甲酸 + 5 mmol/L乙酸铵）,梯度洗脱。负离子模式下多反应监测模式检测。实测6名健康成年人和6只成年SD大鼠样本。  结果  6种初级胆汁酸在6 min内实现基线分离,在5～2 000 μg/L范围内线性良好,相关系数＞0.995,检出限为0.25～0.45 μg/L,定量限为0.84～1.49 μg/L。日内、日间加标回收率分别为86.3 %～111.5 %和87.7 %～104.9 %;日内、日间相对标准偏差分别为5.2 %～11.9 %和6.6 %～14.3 %。健康成年人和成年SD大鼠血浆中初级胆汁酸的浓度显著不同。  结论  该方法简单、快速、灵敏、准确,可用于临床检验和动物试验中初级胆汁酸的快速检测。     

Decreased serum osteocalcin level in non-alcoholic and alcoholic chronic liver diseases

Serum level of osteocalcin (OC) is believed to be a specific biochemical parameter of bone formation. Decreased serum OC has been reported in alcohol-intoxicated subjects, in patients with primary biliary cirrhosis and in patients with chronic alcoholic liver disease. The question was, whether lower OC level could be detected in patients with nonalcoholic and non-cholestatic chronic liver disease. The serum OC was measured by RIA developed in our laboratory. Results were compared to age and sex matched controls. Decreased OC level was found in 35 out of 47 (74%) patients with non-alcoholic and non-cholestatic liver disease as chronic persistent hepatitis, chronic active hepatitis, fatty liver and cirrhosis, in 21 out of 26 (80%) patients with alcoholic liver disease and in 8 out of 15 (53%) primary biliary cirrhosis. None of the patients had elevated value. There was no correlation between the decreased OC level and the duration or severity of the liver disease and the laboratory parameters as bilirubin, AST, ALT, alkaline phosphatase, albumin, prothrombin, and serum 25-OH-D3 vitamin level. Decreased OC was found also in the patients without cirrhosis. The possible causes are discussed. Relying upon these findings it is supposed that chronic liver disease by itself can influence the osteoblast activity also by some unknown mechanism.     

血清总胆汁酸在判断肝损害中的诊断意义

目的评价血清总胆汁酸在各类肝胆疾病中的诊断价值。方法采用BECKMAN CX9全自动生化仪测定健康对照及各类肝胆疾病患者血清中的TBA、ALT、ALP。结果血清总胆汁酸在各种肝病中均有很高的阳性率,TBA阳性率显著高于ALT和ALP。结论血清总胆汁酸对各类肝胆疾病有很高的诊断价值,是肝实质性损伤的灵敏指标。     

Antioxidant therapy in chronic liver diseases

In chronic liver diseases with inflammatory reactions (chronic viral hepatitis, alcoholic liver disease, non-alcoholic steatohepatitis, autoimmune hepatitis, chemicals and drug induced hepatitis, Wilson disease, haemochromatosis) the oxidative stress, the cell alteration induced by free radicals is an important part of the pathogeneis beside the aetiological factors. In this paper the development of free radicals and their physiological role are demonstrated. The factors, which are influenced by the free radicals and antioxidants, as well as the pathological effects caused by oxidative stress in biological systems are discussed in details. The authors demonstrate the free radical processes as key factors in alcoholic liver diseases and non-acoholic steatohepatitis, furthermore the possible treatment modalities in the inhibition of these alterations. Also the free radical processes in chronic viral hepatitis and autoimmune hepatitis are discussed as well. The oxidative stress is able to enhance the progression of chronic inflammatory liver disease independently from the aetiological factor or with it together. That is why the antioxidant drugs could be applied also in the treatment of chronic liver diseases beside the therapy based on the aetiological factors.     

New perspectives in biomonitoring liver function by means of serum bile acids: experimental and hypothetical biochemical basis

The functional activity of the liver and the variety of its responses to injury makes the choice of appropriate tests of function a difficult task. Because of the highly efficient uptake of bile acids by the normal hepatocyte, the determination of serum bile acid (SBA) concentration has been proposed as a test to detect early changes of liver function not associated with cytotoxicity. Several biomonitoring studies have been carried out on subjects occupationally exposed to hepatotoxic substances, by evaluating SBAs as indicators of early liver dysfunction. Even though these studies are not exactly comparable because of the different protocols adopted, most of them show a significant increase in SBA concentrations among the exposed subjects compared with unexposed controls. Furthermore, higher prevalences of subjects with abnormal SBA concentrations occur in those exposed to mixtures of organic solvents. Increased SBA concentrations among the subjects exposed to various xenobiotics have been explained by assuming a change in function of hepatocytes. As regards the nature of the mechanisms involved in the increase in SBA concentrations, recent experimental observations pointed out that some chlorinated aliphatics were able to inhibit cell membrane ATPases and alter cytosolic calcium homeostasis. The lack of any relation, however, between exposure and SBA concentrations remains an important point to clarify and at present prevents the use of measurement of SBA concentrations as an index of effect.     

Importance of serum bile acids determination in adolescents with juvenile hyperbilirubinaemia

The aim of the study was to identify moderate liver impairment in a group of hyperbilirubinaemic adolescents. Using gas chromatography we assessed both total bile acid and primary bile acid levels in 50 adolescents with juvenile hyperbilirubinaemia. At the same time we performed hepatologic examinations and subsequent follow-up assessment of these patients for a period of at least 2 years. As a control group we examined 30 adolescents without any impairment of both the liver and gastrointestinal tract, and 18 patients with low grade (moderately) active chronic hepatitis. In both groups we assessed total and primary bile acids levels as well as conventional liver tests (bilirubin, ALT, AST). On the basis of the clinical course and laboratory findings we divided our patients with juvenile hyperbilirubinaemia into two groups: a group of individuals with Gilbert's syndrome (30 patients) and a group of individuals with probable moderate liver impairment (20 patients). The latter group consisted of the adolescents who exhibited bilirubinaemia over 90 micromol/l and/or exhibited hepatomegaly or splenomegaly proved by the ultrasound examination and/or exhibited intermittent elevation of the liver aminotransferases serum levels. In the group of individuals with moderate liver impairment serum total bile acid levels were significantly elevated in 26% of patients, and the serum cholic acid level was significantly elevated in 25% of patients. These two parameters mutually correlated at a high level of significance. Juvenile hyperbilirubinaemia is one of the common conditions of adolescent age. Its etiology is diverse; it includes both benign conditions like Gilbert's syndrome and post-hepatitic and toxic conditions that require a long-term regimen and follow-up examinations. The number of people suffering from juvenile hyperbilirubinaemia has been growing in the population. Currently 4-6% of the adolescent population suffers from this disease. This growing number is probably caused by external factors of our environment (infection, toxic effects). The determination of mild liver disease in hyperbilirubinaemic patients and the provision of an adequate regimen of exercise and adequate nutritional measures is of great importance for the health of the adolescent population.     

Genetic background of multifactorial liver and bile duct diseases

The majority of liver diseases, are complex. They are the results of interactions between several genes and environmental factors. Familial aggregation and higher concordance rate of monozygotic twins compared to those of dizygotic twins provide evidence for the importance of genetic factors in the pathogenesis. There are only limited data in connection with the genetic background of multifactorial liver diseases. In the future, the genetic background may permit prevention, early, accurate diagnosis, prediction of disease course, complications, prognosis, as well as treatment response.     

New perspectives in biomonitoring liver function by means of serum bile acids: experimental and hypothetical biochemical basis.

The functional activity of the liver and the variety of its responses to injury makes the choice of appropriate tests of function a difficult task. Because of the highly efficient uptake of bile acids by the normal hepatocyte, the determination of serum bile acid (SBA) concentration has been proposed as a test to detect early changes of liver function not associated with cytotoxicity. Several biomonitoring studies have been carried out on subjects occupationally exposed to hepatotoxic substances, by evaluating SBAs as indicators of early liver dysfunction. Even though these studies are not exactly comparable because of the different protocols adopted, most of them show a significant increase in SBA concentrations among the exposed subjects compared with unexposed controls. Furthermore, higher prevalences of subjects with abnormal SBA concentrations occur in those exposed to mixtures of organic solvents. Increased SBA concentrations among the subjects exposed to various xenobiotics have been explained by assuming a change in function of hepatocytes. As regards the nature of the mechanisms involved in the increase in SBA concentrations, recent experimental observations pointed out that some chlorinated aliphatics were able to inhibit cell membrane ATPases and alter cytosolic calcium homeostasis. The lack of any relation, however, between exposure and SBA concentrations remains an important point to clarify and at present prevents the use of measurement of SBA concentrations as an index of effect.