Iron Metabolism And HBV-related Chronic Liver Diseases

Objectives:HBV-related chronic liver diseases is a relatively heavy burden of disease in China.Although the overall research has made significant progress, its exact pathogenesis remainsunclear, and the treatment progress updates slow. Based on the mechanism of iron metabolismin the process of inflammation, cancer and other diseases in a clear, and the increasing studyof its correlation with chronic liver disease caused by hepatitis C, we reflected the role of ironmetabolism in the development of HBV-related chronic liver diseases. This study sought todetect iron metabolism related serological indicators and serum hepcidin levels in differentdevelopment stages of HBV-related chronic liver diseases, and analyzed its correlation withbiochemical and hepatitis B virological and serological indicators of HBV-related chronicliver diseases.Methods:1. Subjects:We chosed totally240patients of HBV-related liver disease in case groupswhich from the First Hospital of Jilin University in2012-2014, including78patients withchronic hepatitis B(CHB group),85patients with liver cirrhosis (LC group) and77patientswith hepatocellular carcinoma (HCC group). And we chosed78healthy persons as normalcontrol group(NC group).2. Detection methods: We measured serum hepcidin levels by using ELISA method, andmeasured iron metabolism related serological indicators for the subjects. The clinical indictorsincluding liver function, Blood routine,coagulation, HBV markers, HBV-DNA loading, alphafetoprotein were detected.3. Statistical analysis: Count data were compared using chi-square test; the normallydistributed of quantitative data were using Kolmogorov-Smirnov test; according to the normaldistribution of the data tested its statistical analysis. We analysed the difference between3 groups or more than3groups with normal distribution of data by using variance, andcompared with LSD method between the two groups in any. We analysed the rest of themeasurement variables by using nonparametric test, using Kruskal-Wallis test among multiplegroups and Mann-Whitney test between any two groups. Using Pearson’s or Spearman’sanalysis to analyse the correlation of the data. We analysed the correlation of hepcidin levelsand other complex variables by using multivariate linear regression analysis. P<0.05wasconsidered to have significant difference, which had statistically significant. The aboveanalysis performed by SPSS22.0software.Results:1.240patients of CLD group (78patients with CHB group,85patients with LC group,77patients with HCC group) and78healthy persons of NC group were detected. By usingKruskal-Wallis test, the results showed that iron metabolism related serological indicatorsdistribution were statistically significant in four groups (serum iron P<0.001, TIBC P<0.001,serum ferritin P<0.001, transferrin saturation P<0.001, serum hepcidin P<0.001, serumtransferrin P<0.001), suggesting that iron metabolism disorders present in patients of HBV-related liver diseases.2. Serum iron levels showed no significant difference between CHD and NC groups(P=0.932). The levels of TIBC, serum ferritin, transferrin saturation, serum hepcidin andserum transferrin were statistically significant between the two groups(P<0.001, P<0.001,P=0.001, P<0.001, P<0.001). Among them, serum ferritin levels in CLD group weresignificantly higher than that in NC group, and its serum hepcidin levels were significantlylower than that in NC group. It is suggested that iron metabolism disorders present in patientsof HBV-related liver disease, with serum iron overload trend. Although serum iron levelswere no significant difference between the two groups, its changes are not the same indifferent development stages of the diseases.3. Serum hepcidin, serum transferrin and TIBC levels in CHB group were significantlylower than that in NC group (P<0.001, P<0.001, P=0.004), suggesting that its regulation ofiron metabolism may be affected by multiple factors as virus itself, the host immune and environment. It contributed to intestinal absorption of iron and macrophages release iron intothe blood, which contributed to increase serum iron levels and promote the formation of ironoverload. The levels of serum iron, serum ferritin and transferrin saturation in CHB groupwere significantly higher than that in NC group(P=0.008, P<0.001, P<0.001), suggesting thatiron overload present in CHB stage which caused by HBV-related chronic liver inflammation,and serum ferritin may be an applied indicator to reflect the liver inflammation in CHB.4. The levels of TIBC, serum hepcidin and serum transferrin in LC group weresignificantly lower than that in NC group(P<0.001, P<0.001, P<0.001). As we noted earlier,decreased hepcidin and transferrin levels promoted serum iron levels increasing, which wasconducive to the formation of serum iron overload. But in LC group, serum iron levelsshowed no significant difference between LC and NC groups (P=0.775) under the conditionof serum ferritin and transferrin saturation significantly elevated. It is suggested that theserum excess iron may be excessive storage in the liver. The ability of liver cells metabolizingiron may decline in liver cirrhosis. So it was difficult to discharge excess iron from liver andthe iron reduced the iron recycle. Then serum iron levels had not a significant increase in thecase of hepcidin significantly reduced. These results indicate that iron metabolism disorderspresent in liver cirrhosis stage and serum ferritin may be a potential clinical indicator toreflect the degree of liver fibrosis. At the same time, iron metabolism disorders’ characteristicswere varied in different development stages of HBV-related liver diseases. After long-termchronic liver inflammation, serum iron overload level by the initial stage of inflammation wasgradually closer to normal. It was the results that body adjusted synthetically under chronicinflammation effect. Although serum iron levels decreased, due to physiological andpathological features of liver cirrhosis, there may be to undertake excessive liver ironoverload in the liver stage of chronic inflammation, which promoted disease progresses.5. Transferrin saturation levels showed no significant difference between HCC and NCgroups (P=0.731). Serum ferritin levels in HCC group were significantly higher than that inNC group (P<0.01). The levels of serum iron, TIBC, serum hepcidin and serum transferrin inHCC group were significantly lower than that in NC group (P<0.001, P<0.001, P<0.001), suggesting that iron metabolism disorders present in patients of HBV-related HCC.Compared with liver cirrhosis, liver iron deposition may be further aggravated in HCC.6. The levels of serum ALT, HBV-DNA loading, serum iron, TIBC, serum hepcidin andserum transferrin were statistically significant between CHB and LC groups (P<0.001,P<0.001, P=0.024, P<0.001, P=0.001, P=0.002). Serum ferritin and transferrin saturationlevels showed no significant difference between the two groups (P=0.932, P=0.637). Amongthem, the levels of serum ALT, HBV-DNA loading, serum iron, TIBC and serum transferrin inLC group were significantly lower than that in CHB group, and its serum hepcidin levels weresignificantly higher than that in CHB group. Compared with LC (HBV-DNA loading wasrelatively low, the inflammatory activity decreased and liver fibrosis increased), hepcidinlevels declined in CHB caused by HBV-DNA loading, abnormal T cell response induced byHBV and inhibited signaling pathways that liver cells produced hepcidin which mediated withrelated cytokines. Although the decline hepcidin levels were beneficial to elevated serum ironlevels, liver cells regeneration and repair ability of damage in CHB was significantly strongerthan liver cirrhosis. Therefore, even if serum iron overload and excessive transport to the liver,the liver had a better ability of its metabolism, which was conducive to iron recycling.7. The levels of HBV-DNA loading, serum iron, TIBC, transferrin saturation, serumhepcidin and serum transferrin were statistically significant between CHB and HCC groups(P<0.001, P<0.001, P=0.011, P=0.004, P=0.038, P=0.032). Serum ferritin levels showed nosignificant difference between the two groups (P=0.141). Among them, the levels ofHBV-DNA loading, serum iron, TIBC and serum transferrin in HCC group were significantlylower than that in CHB group, and its serum hepcidin levels were significantly higher thanthat in CHB group. In addition to the above described mechanism, the tumor cells mayinhibite hepcidin expression.8. The levels of serum iron, TIBC, transferrin saturation and serum hepcidin werestatistically significant between LC and HCC groups (P=0.012, P=0.015, P=0.001, P=0.043).HBV-DNA loading, serum ferritin and serum transferrin levels showed no significantdifference between the two groups (P=0.53, P=0.837, P=0.224). Among them, the levels of serum iron, serum transferrin and serum hepcidin in HCC group were significantly lower thanthat in LC group, and its TIBC levels were significantly higher than that in LC group. It issuggested hepcidin expression was inhibited in patients of HBV-related HCC. Although serumiron levels decreased, the stored iron levels had an increasing trend. It may be related withphysiological and pathological features of liver cancer cells and iron deposition in the liverthat can not be further recycled. Liver iron deposition in HCC group may be higher than thatin LC group due to serum iron levels in HCC group were significantly lower than that in LCgroup.9. Serum hepcidin levels showed no significant difference among mild group, moderategroup and severe group (P=0.676). The levels of serum iron, TIBC, serum ferritin, transferrinsaturation and serum transferrin had significant difference among three groups in CHBpatients(all P<0.001), and the levels of serum iron, serum ferritin and transferrin saturationwere increased with severity of chronic hepatitis B disease. It is suggested iron overload wasrelated with the severity of liver chronic inflammation caused by HBV. Serum ferritin andtransferrin saturation may be applied indicators to reflect the severity of liver inflammation inCHB.10. Serum iron and serum hepcidin levels showed no significant difference among classA, class B and class C groups which were divide according to Child-Pugh grading in patientswith HBV-related cirrhosis (P=0.884, P=0.319). The levels of TIBC, serum ferritin,transferrin saturation and transferrin were significantly different among the three groups(P<0.001, P=0.017, P=0.026, P<0.001). Further more, with the increasing of lesion of liverfunction, the levels of serum ferritin and transferrin saturation showed an up trend, while thelevels of TIBC and transferrin showed a declined tendency. It is suggested with thedeterioration of liver function in patients with liver cirrhosis, iron metabolism disorder wouldfurther aggravated in vivo.11. Serum ferritin and serum hepcidin levels showed no significant difference amongstage A, stage B, stage C and stage D groups which were divide according to Barcelona ClinicLiver Cancer(BCLC) staging in patients with HBV-related HCC(P=0.376, P=0.962). The levels of serum iron, TIBC, transferrin saturation and transferrin were significantly differentamong the four groups (P=0.001, P=0.034, P=0.001, P=0.029).12. The multiple linear regression analysis showed that serum hepcidin levels hadobvious correlation with HBV-DNA loading levels (β=-0.277, t=-4.438, P<0.001), age(β=0.236, t=3.709, P<0.001) and INR (β=0.2, t=2.521, P=0.012) in patients of HBV-relatedliver diseases.13. To further define the relationship between serum hepcidin levels and HBV-DNAloading levels, we performed multiple linear regression analysis for156patients of HBV-related liver diseases without anti-viral therapy. It showed that serum hepcidin levels hadobvious correlation with HBV-DNA loading levels(β=-0.356, t=-4.403, P<0.001) andINR(β=0.688, t=2.424, P=0.017).Conclusions:1. Serum ferritin levels in CLD group were significantly higher than that in NC group,and its serum hepcidin levels were significantly lower than that in NC group. It is suggestedthat iron metabolism disorders present in patients of HBV-related liver disease, with serumiron overload trend.2. Serum hepcidin and ferritin levels may be the sensitive potential clinical indicators toreflect the degree of liver fibrosis and inflammation activity in patients with HBV-relatedchronic liver diseases.3. Iron metabolism disorders’ characteristics were varied in different development stagesof HBV-related liver diseases and regulated by multiple factors.4. High HBV-DNA loading and chronic liver inflammation caused by HBV may reducethe secretion of hepcidin.