© Copyright 2006 by Humana Press tne All nights of any nature, whatsoever, ese (63-4981 /06/ 10901-0515 $30.00 Selenium, Iron, Copper, and Zinc Levels and Copper-to-Zinc Ratios in Serum of Patients at Different Stages of Viral Hepatic Diseases CHING-CHIANG LiN,'? JEE-Fu HUANG,’ LI-Yu Tsa\,* AND YEou-LiH Huanc** Department of Laboratory Medicine, ‘Basic Medical Science Education Center, and ‘Department of Internal Medicine, Fooyin University Hospital, Pingtong, Taiwan; and ‘Faculty of Biomedical Laboratory Science, Kaohsiung Medical University, Kaohsiung, Taiwan Received April 22, 2005; Accepted May 10, 2005 ABSTRACT Viral hepatic diseases, especially those induced by the hepatitis B virus, can progress into more serious pathological outcomes and eventu- ally to hepatocellular carcinoma, A growing body of evidence indicates that many trace elements play important roles in a number of carcinogenic processes that proceed through various mechanisms. To examine the sta- tus of trace elements during the development of hepatic carcinoma, we determined the selenium, iron, copper, and zinc levels and copper-to-zine ratios in the serum of patients at different stages of viral hepatic disease. We observed significant changes in the selenium, iron, copper, and zine levels in the serum of patients having hepatocellular carcinoma, relative to those of healthy controls (p < 0.05). The mean serum copper level in patients with hepatocellular carcinoma was significantly higher than that Of the control group. In contrast, the mean selenium, iron, and zine levels, in patients having hepatocellular carcinoma were significantly lower than those of the control group. In adalition, the mean zine level in the serum of patients with hepatic cirrhosis was significantly lower than that of the con: trol group (p < 0.05). Moreover, we found markedly elevated Cu : Zn ratios. (p < 0.05) in patients having hepatic cirrhosis or hepatocellular carcinoma, (Our findings imply that the levels of some trace elements, such as sel ‘Author to whom all correspondence and reprint requests should be addressed Biological Trace Element Research 15 Vt, 109, 200616 Lin et al. nium, iron, copper, and zinc, and Cu : Zn ratios, might serve as biomark- crs for the increased severity of viral hepatic damage. Index Entries: Selenium; iron; copper; zine; serum; viral hepatic disease, INTRODUCTION Hepatitis B is a serious disease caused by the hepatitis B virus (HBV) infecting the liver, HBV can result in citthosis of the liver, liver cancer, liver failure, and death. Some people who become infected with HBV remain that way for life and can spread the virus to others. About 1.25 million peo- ple living in the United States have life-long hepatitis B (1). Not all people, however, who are infected with HBV look or feel sick; they might posse: the virus and not exhibit any of its symptoms. According to World Health Organization estimates (2), approx 350 million people around the world are long-term (chronic) carriers of the HBV, including more than 2.5 mil- lion people in Taiwan. ‘Taiwan is a hyperendemic area for hepatitis B virus infection (3,4). The prevalence of chronic HBV carriers is relatively high. According to previ- ‘ous reports (5-7), the prevalence rate for hepatitis B carriers in some areas of Taiwan ranges from 15% to 20%. Some of the people who are infected might become carriers, display chronic hepatitis, and progress eventually into cirrhosis or even degenerate into hepatocellular carcinoma (HCC) (8). Accordingly, many residents of Taiwan have the potential to suffer from hepatitis B virus infection. A growing body of evidence indicates that many trace elements play important roles in a number of biological processes through (1) their acti- vating or inhibiting of enzymatic reactions, (2) their competing with other elements and metalloproteins for binding sites, (3) their affecting the per- meability of cell membranes, or (4) other mechanisms (9,10). [tis reasonable, therefore, to assume that these trace elements exert actions—directly or indi- rectly—upon the carcinogenic process (9). Trace elements such as selenium (Ge), iron (Fe), copper (Cu), and Zn exert important protective or enhancing, effects on the progression of some diseases (11). Statistically significant dif- ferences from the Normal distributions of Fe, Cu, Zn, and Se have been reported to occur in patients having various forms of cancer (12,13). An epidemiological study among chronic carriers of HBV in Taiwan indicated that selenium levels in plasma were markedly lower in patients who developed hepatocellular carcinoma than in healthy Hbs\g-positive controls (14). Apart from that epidemiological survey for Se distribution, very few studies have been devoted to the investigation of the status of trace elements in patients at different stages of hepatitis B virus infection. To understand the possible association between the status of some trace ele- ments and the development of viral hepatic disease, in this study we exam- ined the serum Se, Fe, Cu, and Zn levels and Cu-to-Zn ratios in patients Biological Trace Element Research Vol. 109, 2006Serum Element Levels in Liver Viral Diseases 17 having chronic hepatitis, hepatic cirthosis, and hepatocellular carcinoma and we compared these levels with those of healthy control subjects. MATERIALS AND METHODS Subjects This study included one group of healthy control subjects, three groups of patients diagnosed with different extents of hepatic diseases (chronic hepatitis B, hepatic cirrhosis, and hepatocellular carcinoma), and one group of carriers that was positive for HbsAg. None of the participants had taken mineral supplements before blood sampling. All of the subjects provided their informed consent as approved by the local medical ethics committee. We considered the subjects that were HbsAg positive or anti-HCV (hepatitis C virus) positive for more than 6 mo yet had normal liver function and normal clinical findings to be asymptomatic carriers. Ifthe subjects were HbsAg positive or anti-HCV positive and had abnormal liver function for 6 mo, then we considered them as patients having chronic hepatitis B. We con- sidered the patients who were HbsAg positive and/or anti-HCV positiv. had abnormal liver function and abnormal laboratory findings, and dis- played abnormal abdominal ultrasonographs to have cirrhosis. Our diag- noses of hepatocellular carcinoma were based on a positive finding from pathologic or cytological examinations, We chose control subjects from those who were HbsAg and anti-HCV negative, displayed no systemic diseases, and had normal mean corpuscular volume and liver function. Blood was drawn by venipuncture into Venoject tubes in the morning after an overnight fast. Part of the serum was stored at -70°C until it was needed for metal analysis; the remainder was used for all of the laboratory measurements. Table 1 lists the clinical and biochemical features of the subjects investigated. Serological Viral Markers and Biochemical Tests Serum HbsAg was detected using a second-generation enzyme immunoassay kit (Abbott Laboratories, North Chicago, IL, USA). Anti- HCV was measured by means of a third-generation enzyme immunoassay kit (Abbott Laboratories, North Chicago, IL, USA). Serum albumin, globu- lin, total bilirubin, aspartate aminotransferase (AST), and alanine amino- transferase (ALT) were measured using an automatic analyzer (Hitachi Mode! 7060), Serum Trace Elements Determination A Perkin-Elmer 5100PC atomic absorption spectrophotometer equipped with an HGA 600 graphite furnace programmer was used to Biological Trace Element Research Vt. 109, 2006