Comparison of hepatic oxidative DNA damage in patients with chronic hepatitis B and C

Summary.  8-Hydroxydeoxyguanosine (8-OHdG) is a promutagenic DNA lesion produced by hydroxyl radicals and is recognized as a useful marker in estimating DNA damage induced by oxidative stress. The aim of this study was to clarify the clinical significance of hepatic 8-OHdG levels in patients with chronic viral hepatitis. Hepatic 8-OHdG accumulation was investigated in patients with chronic hepatitis C (CH-C) ( n  = 77) and chronic hepatitis B (CH-B) ( n  = 34) by immunohistochemical staining of liver biopsy samples. 8-OHdG positive hepatocytes were significantly higher in patients with CH-C compared to CH-B (median 55.0 vs 18.8 cells/10⁵ μm², P  < 0.0001). The number of positive hepatocytes significantly increased with the elevation of serum aminotransferase levels, especially in CH-C patients (8-OHdG vs alanine aminotransferase (ALT)/aspartate aminotrasferase (AST) were r  = 0.738/0.720 in CH-C and 0.506/0.515 in CH-B). 8-OHdG reactivity was strongly correlated with body and hepatic iron storage markers in CH-C ( vs serum ferritin, r  = 0.615; vs hepatic total iron score, r  = 0.520; vs hepatic hepcidin mRNA levels, r  = 0.571), although it was related to serum HBV-DNA titers ( r  = 0.540) and age of patients ( r  = –0.559) in CH-B. These results indicate that hepatic oxidative DNA damage is common in chronic viral hepatitis, in particular chronic HCV-infected patients, suggesting a possible link between chronic hepatic inflammation and hepatocarcinogenesis. The strong positive correlation between hepatic DNA damage and iron overload suggests that iron content is one of the most likely mediators of hepatic oxidative stress and iron reduction may be beneficial to reduce the incidence of hepatic cancer in CH-C patients.     

慢性丙型肝炎的组织学特点和分类

目的研究慢性丙型肝炎的组织病理学特点和新的病变程度的分类，并与慢性乙型肝炎作比较。方法对４９例慢性丙型肝炎和４５例慢性乙型肝炎的肝组织标本，由三位病理科医师双盲读片，记录主要的组织学改变，并根据新老慢性肝炎分类评估。结果慢性丙型肝炎最为突出的组织学改变为肝脂肪变（６１％）、胆管损害（５７％）、肝窦内炎症细胞（４７％）和淋巴细胞聚集（４３％），均较慢性乙型肝炎多见，差异有显著性。本组病例按新的国际分级、分期与传统的慢活肝、慢迁肝的分类比较，二者之间有一定的相关性，但前者更能准确反映病变的程度。结论新的慢性肝炎的组织学分类法优于传统的分类法。值得推广。     

Upregulation of transferrin receptor 2 and ferroportin 1 mRNA in the liver of patients with chronic hepatitis C

BACKGROUND: Iron accumulation has been reported to be associated with progression of liver injury. The mechanism of iron accumulation in the liver is not known. In the present study, hepatic messenger RNA (mRNA) expression of transferrin receptor (TfR)1, TfR2, and ferroportin (FP)1 was measured in patients with chronic hepatitis (CH). METHODS: Eleven patients with CH-B and 43 patients with CH-C were enrolled. All patients underwent liver biopsy. Hepatic expression of TfR1, TfR2 and FP1 mRNA was analyzed using a real-time polymerase chain reaction. Total hepatic iron score (THIS) was evaluated by Prussian blue staining. RESULTS: Serum ferritin concentration is significantly higher in CH-C than in CH-B. Values of THIS of >/=5 were observed only in CH-C patients (44% of CH-C patients). The expression level of TfR2 mRNA was 10-26-fold higher than the TfR1 mRNA expression level. The TfR2 and FP1 mRNA expression was significantly higher in CH-C than in CH-B patients. Hepatic expression of TfR2 and FP1 mRNA was well correlated with THIS. CONCLUSIONS: Hepatic iron accumulation is more severe in patients with CH-C. Upregulation of hepatic iron transporters may contribute to the hepatic iron accumulation in CH-C.     

Treatment with spironolactone for 24 weeks decreases the level of matrix metalloproteinases and improves cardiac function in patients with chronic heart failure of ischemic etiology

BACKGROUND:

The myocardial extracellular matrix is believed to be central to the remodelling that takes place following myocardial infarction. The contribution of markers of collagen metabolism to this process remains less well understood. The present study examined the contribution of some of the markers of collagen metabolism in cardiac remodelling, as well as the effect of spironolactone on the remodelling process.

OBJECTIVES:

To investigate the pathological contribution of markers of collagen metabolism, including matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), type I collagen carboxyterminal telopeptide (ICTP) and procollagen type I carboxyterminal propeptide (PICP), in cardiac remodelling following ischemic cardiomyopathy, and to examine the pharmacoregulatory effects of spironolactone on collagen metabolism.

METHOD:

Eighty-six consecutive patients (62 men and 24 women) with chronic heart failure of ischemic etiology (patient group) and 25 age-matched controls were enrolled in the study. The subjects in the patient group were randomly assigned into a spironolactone or nonspironolactone group. Plasma levels of MMP-9, TIMP-1, ICTP and PICP were measured using ELISA and radioimmunoassay techniques. Furthermore, left ventricular diastolic diameter and ejection fraction were assessed using two-dimensional and Doppler echocardiography.

RESULTS:

The plasma concentrations of MMP-9, TIMP-1 and the MMP-9 to TIMP-1 ratio, as well as ICTP, were significantly increased in the patient group. The PICP to ICTP ratio in the patient group was significantly lower than that in the age-matched control subjects. After a follow-up period of 24 weeks, the PICP to ICTP ratio increased, and MMP-9, TIMP-1 and the MMP-9 to TIMP-1 ratio decreased in the spironolactone subgroup.

CONCLUSIONS:

Biomarkers of collagen degradation were elevated and correlated with depressed heart function; spironolactone may partially reverse the dysregulation in collagen metabolism.     

Collagen metabolism in extracellular matrix may be involved in arterial stiffness in older hypertensive patients with left ventricular hypertrophy.

Collagen metabolism in the extracellular matrix (ECM) is related to the pathogenesis of cardiovascular stiffness and remodeling in hypertension. We evaluated the association between collagen metabolism markers and the newly developed parameter, brachial-ankle pulse wave velocity (baPWV), in older hypertensive patients with left ventricular hypertrophy (LVH). We performed echocardiography and baPWV measurement using a new device, form PWV/ABI (Colin Medical Technology, Komaki, Japan), and measured plasma levels of markers of collagen metabolism such as procollagen type I C-terminal propeptide (PICP: a marker of collagen synthesis), collagen type I pyridinoline cross-linked C-terminal telopeptide (ICTP: a marker of collagen type I degradation), matrix metalloproteinase-1 (MMP-1: a marker of collagen degradation) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in 46 hypertensive patients with LVH. BaPWV was correlated with the plasma level of PICP (r=0.33, p=0.03) and ICTP (r=0.29, p=0.05) and the total TIMP-1/MMP-1 ratio (an index of collagen turnover; r=0.30, p=0.04). BaPWV was negatively correlated with the E/A ratio of left ventricular inflow (r=-0.36, p<0.05), while baPWV was not correlated with left ventricular mass index (LVMI; r=-0.175, p=0.25) or deceleration time of the mitral E wave (DCT; r=0.15, p=0.31). The measures of hypertensive heart disease, such as the E/A ratio, DCT or LVMI were not correlated with any collagen markers in this study. In multiple regression analysis adjusted for confounding factors such as age, sex, pulse pressure, mean blood pressure, pulse rate, LVMI, E/A ratio and DCT, the positive correlation between baPWV and total TIMP-1/MMP-1 ratio remained significant (p<0.05). In conclusion, arterial stiffness in high-risk older hypertensive patients may involve ECM collagen metabolism.     

Effect of lamivudine treatment on plasma levels of transforming growth factor β1, tissue inhibitor of metalloproteinases-1 and metalloproteinase-1 in patients with chronic hepatitis B

AIM: Transforming growth factor (TGF)-β1, metalloproteinase (MMP)-1 and its tissue inhibitor (TIMP)-I are considered predictive biomarkers of chronic hepatitis activity and fibrosis.The aim of this study was to evaluate the effect of lamivudine treatment on the plasma levels of these peptides in patients with chronic hepatitis B.METHODS: TGF-β1, MMP-1 and TIMP-1 plasma concentrations were measured with an enzyme immunoassay in 40 patients treated with lamivudine for 48 wk. Elimination of HBV-DNA and HBV antigens was evaluated 24 wk after treatment completion.RESULTS: Baseline TGF-β1(29.6&#177;2.2 ng/mL) and TIMP-1(1 578&#177;93 ng/mL) significantly exceeded normal values(18.3&#177;1.6 ng/mL and 1 102&#177;67 ng/mL respectively). Lamivudine treatment resulted in a significant decrease of TGF-β1 and TIMP-1 during treatment with an increase after 24 wk of treatment. Pretreatment MMP-1 levels (6.7&#177;0.7 ng/mL) were significantly lower than normal values (11.9&#177;0.9 ng/mL) and increased during treatment and follow-up. A significant correlation was noted between TGF-β1 or TIMP-1 and aminotransferases as well as fibrosis scored in liver biopsy specimens. There were no statistically significant differences of TGF-β1, TIMP-1 and MMP-1 between four groups at baseline, 24 and 48 wk of treatment. TGF-β1 and TIMP-1 levels increased significantly in non-responders and normalized in responders at wk 72. MMP-1 also normalized in responders and decreased to values significantly lower than normal in non-responders.CONCLUSION: These findings support the role of TGF-β1,TIMP-1 and MMP-1 in the pathogenesis of chronic hepatitis B.Because of their association with hepatic injury and antiviral treatment efficacy, determination of these peptides may be useful in disease management.     

Serum thioredoxin elucidates the significance of serum ferritin as a marker of oxidative stress in chronic liver diseases

BACKGROUND/AIMS: Serum thioredoxin (TRX) levels have recently been established as an indicator of oxidative stress in various diseases. The aim of the present study was to clarify the clinical significance of serum ferritin in chronic liver diseases. METHODS: Levels of ferritin, transferrin saturation (TS), aspartate aminotransferase (AST), and TRX were measured in the sera of patients with chronic hepatitis C (CH-C, n=92), chronic hepatitis B (CH-B, n=28), nonalcoholic fatty liver (FL, n=31), or alcoholic liver diseases (ALD, n=17). Serum TRX levels were evaluated with a recently established sandwich enzyme-linked immunosorbent assay kit. RESULTS: Serum TRX levels were significantly higher in CH-C, FL, and ALD than in healthy volunteers. A larger proportion of patients with CH-C, FL, and ALD had elevated levels of serum ferritin than CH-B. Serum ferritin levels were positively correlated with levels of TS, AST, and TRX in CH-C, but were merely correlated with TS values in CH-B. Ferritin levels were also well correlated with AST and TRX, but not with TS in FL and ALD. CONCLUSION: Oxidative stress, which was evaluated by measuring serum TRX, in addition to storage iron and hepatocyte damage is a cause of increasing serum ferritin levels in chronic liver diseases. An elevated serum ferritin level, which was correlated with TS, indicates that iron-induced oxidative stress contributes to CH-C. Elevated ferritin levels in FL and ALD may be mostly due to iron-unrelated stresses.     

Intrahepatic status of regulatory T cells in autoimmune liver diseases and chronic viral hepatitis.

Aim: Regulatory T cells (Tregs) maintain immunological tolerance and suppress autoreactive immune responses. We evaluated the intrahepatic status of Tregs in patients with autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC), chronic hepatitis C (CH-C), or chronic hepatitis B (CH-B). Methods: We analyzed 85 patients (20 AIH, 22 PBC, 27 CH-C, and 16 CH-B) and 14 controls. Using liver tissue samples obtained by needle biopsy or from marginal parts of resected metastatic liver tumors in the controls, immunohistochemical analyses of forkhead box P3(+), which is a specific marker for Tregs, CD4(+), and CD8(+) cells were performed. Results: Intrahepatic Tregs were significantly more infiltrated in patients with liver diseases than in the controls. There were significantly fewer intrahepatic Tregs in the AIH patients than in the PBC patients (P = 0.037). Patients with alow frequency of intrahepatic Tregs were detected significantly more in the AIH and CH-B groups than in the PBC and CH-C groups (P < 0.05). In addition, the frequency of Tregs decreased in the liver of PBC patients as the pathological stage of the disease advanced. We found significantly less infiltration of CD4(+) T cells in AIH than in other diseases (P < 0.05). Liver-infiltrating CD8(+) T cells were detected more frequently in the CH-B group than in other groups (P < 0.003). Conclusion: Intrahepatic Tregs were increased in both patients with autoimmune liver diseases and those with viral hepatitis. In autoimmune liver diseases, intrahepatic Tregs were fewer in the AIH patients than in the PBC patients.     

Elevated serum markers of collagen degradation in patients with mild to moderate dilated cardiomyopathy

BACKGROUND AND AIMS: Left ventricular (LV) dilation and myocardial remodelling are hallmarks of heart failure in idiopathic dilated cardiomyopathy (DCM). Interstitial collagen is essential for LV integrity and function while degradation of collagen by collagenases, especially matrix-metalloproteinases (MMPs), are suggested to contribute to ventricular dilation. In the present study, serological markers of collagen metabolism were investigated. METHODS AND RESULTS: Serum levels of MMP-1 and its inhibitor (TIMP-1), the markers for collagen degradation type I (collagen carboxyterminal telopeptide (ICTP)) and synthesis (carboxyterminal propeptide of type I procollagen (PICP)) were quantified by ELISA and RIA of 43 patients with DCM and 47 age-matched control subjects. Free MMP-1 serum concentration was significantly increased in the DCM group (5.29+/-0.83 vs. 2.22+/-0.29 ng/ml; P=0.01) as well as the free TIMP-1 concentration (206.54+/-12.65 vs. 181.44+/-8.55 ng/ml; P=0.05). The free MMP-1/TIMP-1-ratio was higher in DCM than in the control group (0.030+/-0.005 vs. 0.012+/-0.001; P=0.01). ICTP was significantly increased (7.60+/-1.21 vs. 3.44+/-0.19 microg/l; P<0.001). PICP was not significantly increased (125.29+/-8.93 microg/l vs. 113.11+/-5.47 microg/l; P=n.s.). Free MMP-1 and MMP-1/TIMP-1-ratio correlated with LV end diastolic diameter [cm/m(2) body surface area (BSA)] (r=0.28; P=0.03 and r=0.34; P=0.01, respectively) as well as with cardiac index (CI) (r=-0.32; P=0.04 and r=-0.33; P=0.04, respectively) in patients with DCM. CONCLUSION: Serum markers of collagen degradation are elevated and might be valuable markers for progression of LV dilation in patients with DCM.     

Diagnostic value of platelet derived growth factor-BB, transforming growth factor-β1,matrix metalloproteinase-1, and tissue inhibitor of matrix metaUoproteinase-1 in serum and peripheral blood mononuclear cells for hepatic fibrosis

AIM: Noninvasive diagnosis of hepatic fibrosis has become the focus because of the limited biopsy, especially in the surveillance of treatment and in screening hepatic fibrosis. Recently, regulatory elements involved in liver fibrosis, such as platelet derived growth factor-BB (PDGF-BB), transforming growth factor-β1(TGF-β1), matrix metalloproteinase-1 (MMP-1), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), have been studied extensively. To determine whether these factors or enzymes could be used as the indices for the diagnosis of hepatic fibrosis, we investigated them by means of receiver operating characteristic (ROC) curve. METHODS: Serum samples from sixty patients with chronic viral hepatitis B and twenty healthy blood donors were assayed to determine the level of PDGF-BB, TGF-β1, MMP-1, and TIMP-1 with ELISA, and HA, PCIII, C-IV, and LNlevel with RIA. The message RNA (mRNA) expression of TIMP-1 and MMP-1 in peripheral blood mononuclear cells (PBMCs) was detected by RT-PCR and Northern blot hybridization. Liver biopsy was performed in all patients. The biopsy samples were histopatholocjically examined. The trial was double-blind controlled. RESULTS: The serum level of PDGF-BB, TIMP-1, the ratio of TIMP-1 and MMP-1 (TIMP-1/MMP-1), mRNA expression of TIMP-1 (TIMP-lmRNA), and the ratio of TIMP-lmRNA and MMP-lmRNA (TIMP-lmRNA/MMP-lmRNA) in patients was significantly higher than those in the healthy blood donors (t=2.514-11.435, P=0.000-0.016). The serum level of PDGF-BB, TIMP-1, TIMP-1/MMP-1, and TIMP-lmRNA was positively correlated with fibrosis stage and inflammation grade (r=0.239-0.565, P=0.000-0.033), while the serum level of MMP-1 was negatively correlated with fibrosis stage and inflammation grade, and TIMP-lmRNA/MMP-lmRNA was positively correlated with inflammation grade. Through the analysis by ROC curve, serum PDGF-BB was the most valuable marker, and its sensitivity was the highest among the nine indices. The markers with the highest specificity were TIMP-lmRNA and TIMP-lmRNA/MMP-lmRNA in PBMCs. The area under the curve (AUC) of PDGF-BB, TIMP-lmRNA, TIMP-lmRNA/MMP-lmRNA, TIMP-1/MMP-1, HA, PCIII, TIMP-1, C-IV, and LN was 0.985, 0.876, 0.792, 0.748, 0.728, 0.727, 0.726, 0.583, and 0.463, respectively. The sensitivity and the specificity in the parallel test was 99.0% and 95.0 % when serum PDGF-BB, TIMP-lmRNA and TIMP-lmRNA/MMP-lmRNA was detected simultaneously. CONCLUSION: Serum level of PDGF-BB, TIIVIP-lmRNA, TIMP-lmRNA/MMP-lmRNA in PBMCs, and serum level of TIMP-1 and TIMP-1/MMP-1 can be used as the indices for the diagnosis of hepatic fibrosis, but the former three are more useful. The combination of serum PDGF-BB, TIMP-lmRNA and TIMP-lmRNA/MMP-lmRNA in PBMCs is even more efficient in screening liver fibrosis.     

Increased degradation of type I collagen in patients with inflammatory bowel disease.

To assess the mechanisms of osteopenia in inflammatory bowel disease (IBD), the serum markers of bone formation (osteocalcin and carboxyterminal propeptide of type I procollagen (PICP)) and bone degradation (carboxyterminal telopeptide of type I collagen (ICTP)), the bone mineral density (BMD) of the lumbar spine and the proximal femur and calcium intake of 150 unselected IBD patients and 73 healthy controls were investigated. The patients had higher ICTP values (3.69 (SD 1.40) microgram/l) than the healthy controls (3.25 (1.00) microgram/l, p = 0.035), but no differences in serum PICP and osteocalcin between these groups were detected. In the patients, the ICTP, PICP, and osteocalcin values did not have any significant correlation with BMD, but the patients with ICTP values above 3.6 microgram/l had significantly lower Z scores than those with lower ICTP. In the controls, however, a positive correlation between serum ICTP and BMD was found. The ulcerative colitis patients with total colitis had higher values of ICTP (3.96 (1.58) microgram/l) than those with a left sided disease (3.04 (0.86) micrograms/l, p = 0.009). The patients with a history of clinically active disease (n = 20) had higher ICTP (4.58 (1.55) microgram/l) and osteocalcin (12.56 (5.64) microgram/l) values than the patients (n = 130) with quiescent disease (ICTP 3.56 (1.33), p = 0.002, and osteocalcin 9.76 (3.62), p = 0.017). Increased serum osteocalcin, PICP, and ICTP concentrations and reduced BMD Z scores were found in a subgroup of Crohn's disease patients with a history of an active disease (n = 11). Raised serum ICTP and normal values of osteocalcin and PICP in IBD patients show increased breakdown of type I collagen without a compensatory increase in its synthesis suggesting an increased rate of bone degradation as a probable mechanism for osteopenia in IBD. Raised ICTP values are related to reduced bone mineral densities.     

Different signaling pathways in the livers of patients with chronic hepatitis B or chronic hepatitis C

The clinical manifestations of chronic hepatitis B (CH-B) and chronic hepatitis C (CH-C) are different. We previously reported differences in the gene expression profiles of liver tissue infected with CH-B or CH-C; however, the signaling pathways underlying each condition have yet to be clarified. Using a newly constructed cDNA microarray consisting of 9614 clones selected from 256,550 tags of hepatic serial analysis of gene expression (SAGE) libraries, we compared the gene expression profiles of liver tissue from 24 CH-B patients with those of 23 CH-C patients. Laser capture microdissection was used to isolate hepatocytes from liver lobules and infiltrating lymphoid cells from the portal area, from 16 patients, for gene expression analysis. Furthermore, the comprehensive gene network was analyzed using SAGE libraries of CH-B and CH-C. Supervised and nonsupervised learning methods revealed that gene expression was correlated more with the infecting virus than any other clinical parameters such as histological stage and disease activity. Pro-apoptotic and DNA repair responses were predominant in CH-B with p53 and 14-3-3 interacting genes having an important role. In contrast, inflammatory and anti-apoptotic phenotypes were predominant in CH-C. These differences would evoke different oncogenic factors in CH-B and CH-C. In conclusion, we describe the different signaling pathways induced in the livers of patients with CH-B or CH-C. The results might be useful in guiding therapeutic strategies to prevent the development of hepatocellular carcinoma in cases of CH-B and CH-C.     

巨噬细胞移动抑制因子与慢性乙型肝炎、乙型肝炎肝硬化的关系

目的 了解慢性乙型肝炎(乙肝)及乙肝肝硬化患者血清细胞因子巨噬细胞移动抑制因子(MIF)表达水平,探讨血清MIF水平与Ⅲ型前胶原肽(PⅢP)、组织金属蛋白酶抑制剂-1(TIMP-1)的相关性.方法 选择44例慢性乙肝患者(肝炎组)、44例乙肝肝硬化患者(肝硬化组)和30名健康者(对照组),取静脉血并应用ELISA方法检测其血清中细胞因子MIF浓度,分析实验组血清MIF水平与PⅢP、TIMP-1相关性.两组间均数比较采用t检验,MIF水平与ALT、AST、TBil、PTA、PⅢP及TIMP-1关系采用直线相关分析.结果 慢性乙肝组及肝硬化组血清MIF、PⅢP及TIMP-1的浓度比健康对照组升高.差异均有统计学意义(t=12.87、5.28、10.98,t=11.22、14.84、11.17)均P<0.05),但该3种细胞因子在慢性乙肝及肝硬化患者之间差异无统计学意义(t=-1.05、1.52、-2.07;均P>0.05);MIF水平与ALT、AST、TBil及PTA均无相关性;HBeAg阳性和高病毒载量(>1×105拷贝/mL)慢性乙肝患者体内MIF水平高于HBeAg阴性和低病毒载量(≤1×105拷贝/mL)患者.MIF水平与PⅢP水平在慢性乙肝组和肝硬化组中均存在正相关(r=0.603,P<0.05;r=0.415,P<0.05);MIF水平与TIMP-1水平在慢性乙肝组中存在正相关(r=0.458,P<0.05),而在肝硬化组无相关性(r=0.210,P>0.05),PⅢ P与TIMP-1在慢性乙肝组及肝硬化组均存在相关性(r=0.649,P<0.05)r=0.424,P<0.05).结论 慢性乙肝及肝硬化患者体内MIF水平明显升高,MIF的早期生成可能与病毒复制有关,与肝功能无关;MIF参与了肝炎,肝纤维化及肝硬化形成过程,可反应肝硬化程度.     

Plasma Levels of Matrix Metalloproteinase-2 (MMP-2) and Tissue Inhibitors of Metalloproteinases-1 and -2 (TIMP-1 and TIMP-2) as Noninvasive Markers of Liver Disease in Chronic Hepatitis C Comparison Using ROC Analysis

As chronic liver disease progresses, animbalance occurs between synthesis and breakdown ofextracellular matrix (ECM). Matrix metalloproteinases(MMPs) are involved in degrading ECM while tissueinhibitors of metalloproteinases (TIMPs) prevent theirfibrolytic action. We determined if plasma levels ofMMP-2, TIMP-1 and TIMP-2 are related to liver histologyin patients with chronic hepatitis C. Plasma MMP-2, TIMP-1 and TIMP-2 levels were measured in 43patients with chronic hepatitis C. Plasma levels ofMMP-2, TIMP-1 and TIMP-2 and serum ALT were correlatedwith liver biopsy score and specificity and sensitivity of the assays in detecting advanced liverdisease were calculated from ROC analysis. Plasma TIMP-1was significantly correlated with histological activityindex (r = 0.45), portal inflammation (r = 0.48), periportal necrosis (r = 0.34) and focalnecrosis (r = 0.38). Plasma TIMP-2 was significantlycorrelated with fibrosis (r = 0.43) and confluentnecrosis (r = 0.41). Using ROC analysis both TIMP-1 andTIMP-2 had significant diagnostic ability in detectingadvanced liver disease (Area under the curve 0.73 forboth, p 0.015 and 0.036 respectively). A normal plasmaTIMP-1 excluded advanced liver disease. Neither plasma MMP-2 or serum ALT were related tofibrosis or to histological activity index. Withincreased severity of liver disease in chronic hepatitisC there is increased plasma levels of TIMPs-1 and-2.Plasma TIMP-1 and TIMP-2 are sensitive and to a lesserextent specific in detecting advanced liver disease inchronic hepatitis C and could be used in preference toserum ALT.     

Regulation of circulating immune complexes by complement receptor type 1 on erythrocytes in chronic viral liver diseases

BACKGROUND AND AIM: Complement receptor type 1 (CR1) is a transmembrane protein, and human erythrocyte CR1 (E-CR1) is involved in the transport of circulating immune complexes (IC) from the circulation to the reticuloendothelial system, including the liver and spleen. In chronic viral hepatitis, increased levels of IC containing viral particles and an association with various extrahepatic manifestations have been reported. However, regulatory mechanisms for IC levels are not fully understood. PATIENTS/SUBJECTS AND METHODS: We analysed IC, E-CR1, and quantitative polymorphism of the CR1 gene in 149 patients with chronic viral liver diseases and in 64 normal blood donors using an enzyme linked immunosorbent assay, radioimmunoassay, and polymerase chain reaction-restriction fragment length polymorphism, respectively. We also analysed the effect of CR1 gene polymorphism on IC binding to E-CR1 using molecular methods. RESULTS: E-CR1 levels in patients with chronic hepatitis and chronic viral liver diseases as a whole correlated inversely with increased levels of IC. Moreover, significantly high levels of IC were observed in patients with chronic hepatitis C (CH-C) who were homozygous for the E-CR1 low density allele. We also found low levels of E-CR1 in liver cirrhosis and CH-C but not in CH-B. Low levels of E-CR1 in CH-C were observed, even after considering the polymorphism of the CR1 gene. Finally, we demonstrated CR1 gene polymorphism dependent binding of hepatitis virus containing IC. CONCLUSIONS: Our results emphasise the important role of E-CR1 in clearance of IC from the circulation and the acquired, rather than inherited, decrease in E-CR1 in chronic viral liver diseases, especially of type C.     

慢性乙型肝炎患者肝组织基质金属蛋白酶组织抑制物-1表达及其意义

目的：分析慢性乙型肝炎患者血清和肝组织基质金属蛋白酶组织抑制物-1（TIMP-1）表达水平与肝纤维化分期的相关性。方法分别采用免疫组化法和双抗体夹心酶联免疫吸附法检测患者肝组织和血清 TIMP-1表达水平,分析两者相关性;对159例慢性乙型肝炎患者行肝活检病理学检查,以组织学活动指数（HAI）予以分级（G）和分期（S）,分析慢性乙型肝炎患者血清 TIMP-1水平与 HAI 分级和纤维化分期的相关性。结果慢性乙型肝炎患者肝组织 TIMP-1表达水平与其血清水平呈显著性正相关（r=0.9521,P＜0.01）;不同肝纤维化分期（S1～S4）慢性乙型肝炎患者血清 TIMP-1水平差异显著（P＜0.05）,且血清 TIMP-1水平与肝纤维化分期呈正相关（r=0.704, P＜0.01）,但不同炎症分级的慢性乙型肝炎患者血清 TIMP-1水平差异无显著性。结论血清 TIMP-1水平可以较好地反应肝脏纤维化程度,且不受肝组织炎症分级的影响,有望被用于肝组织纤维化程度的临床无创评估。     

Successful interferon therapy reverses enhanced hepatic iron accumulation and lipid peroxidation in chronic hepatitis C

OBJECTIVES: Hepatic iron deposition has been reported in chronic hepatitis C (CH-C), and iron-induced lipid peroxidation may be involved in the pathogenesis of CH-C. The aims of the present study were: 1) to determine whether patients with CH-C have evidence of enhanced hepatic lipid peroxidation and to evaluate its relation to iron status, compared with that in patients with chronic hepatitis B (CH-B); and 2) to assess the effect of interferon (IFN) therapy on hepatic iron and lipid peroxidation. METHODS: In the liver biopsies of 40 patients with CH-C and 26 patients with CH-B, immunohistochemical detection of 4-hydroxy-2-nonenal (HNE)-protein adducts for evaluation of lipid peroxidation was performed, and hepatic iron status was biochemically and histologically assessed. In 16 CH-C patients with normal serum transaminases and undetectable serum HCV-RNA >6 months after the end of IFN treatment (responders) and in 11 nonresponders, hepatic HNE-protein adducts and siderosis were evaluated in pre- and posttreatment liver biopsies. RESULTS: Hepatocytic HNE-protein adducts and iron deposits were more abundant in the patients with CH-C than in those with CH-B. No correlation was found between the levels of hepatocytic HNE-protein adducts and hepatic iron status in either of the two groups. In the responders to IFN treatment for CH-C, hepatocytic HNE-protein adducts disappeared or attenuated with improvement of hepatic siderosis after the treatment, whereas IFN treatment did not improve hepatocytic expression of HNE-protein adducts and hepatic siderosis in the nonresponders. CONCLUSIONS: Patients with CH-C have evidence of enhanced hepatic iron accumulation and lipid peroxidation compared to those with CH-B. In CH-C, hepatic siderosis and lipid peroxidation are improved with successful IFN treatment. These results suggest that hepatic lipid peroxidation and iron may potentially play contributory roles in the pathogenesis of CH-C.     

慢性乙型肝炎肝组织中MMP-2、TIMP-1与肝纤维化关系的研究

为了研究慢性乙型肝炎患者肝纤维化与肝组织中基质金属蛋白酶-2(MMP-2)及其天然抑制物金属蛋白酶组织抑制剂-1(TIMP-1)的关系,对60例慢性乙型肝炎患者进行肝组织病理活检,其中S_110例,S_224例,S_312例,S_414例。采用苦叶酸天狼猩红染色检测胶原面积百分比,单克隆抗体(McAb)免疫组织化学染色检测MMP-2、TIMP-1阳性细胞的方法进行病理分析。结果显示：慢性乙型肝炎肝组织中胶原面积百分比与肝组织纤维化分期正相关(r=0．885,P=0．000);慢性乙型肝炎肝组织中MMP-2与肝纤维化的分期无关(r=0．034,P= 0．896);慢性乙型肝炎肝组织中TIMP-1和肝纤维化的分期正相关(r=0．760,P=0．000);慢性乙型肝炎肝组织中MMP-2/TIMP-1和肝纤维化分期负相关(r=-0．674,P=0．000)。总之,TIMP-1参与了慢性乙型肝炎肝纤维化纤维化的过程,而MMP-2/TIMP-1是诊断肝纤维化的比较合适的指标。     

Characteristic Laparoscopic Findings of Chronic Hepatitis C: A Comparison with Hepatitis B.

Abstract: We performed laparoscopy on 72 patients with chronic hepatitis C (CH-C) to elucidate its morphological features and characteristics relating to its progressive clinical course. We then compared these findings with those of 188 patients with chronic hepatitis B (CH-B). The frequency of reddish hepatic markings was almost the same in the patients with CH-B (21%) and the patients with CH-C (24%). In the patients with CH-B, reddish markings were present more frequently in the subjects with sublobular hepatic necrosis (34%) than in those without it (5%, p<0.01), but in CH-C the difference was not significant. Reddish markings were more even in the patients with CH-B with respect to form, size and distribution, whereas those in the patients with CH-C were more uneven. Prenodular patchy markings were present more frequently in the patients with CH-B (14%) than in the patients with CH-C (6%, p<0. 05). These findings suggested that the progression of hepatic necrosis was more variable in different parts of the liver, and that the regeneration of hepatocytes was less active in the patients with CH-C, compared with the patients with CH-B.     

TIMP-1: a marker of left ventricular diastolic dysfunction and fibrosis in hypertension

This study was designed to document noninvasively the pathological mechanisms responsible for myocardial fibrosis and to assess the clinical utility of plasma markers of collagen synthesis and degradation as screening tools for the assessment of fibrosis in hypertension. We studied 100 never-treated hypertensive patients and 50 normal subjects. Echocardiographic assessment was made of left ventricular (LV) mass and diastolic filling using measurement of E:A ratio, E wave deceleration time (E dec), and isovolumic relaxation time (IVRT). The presence of diastolic dysfunction was taken as a surrogate marker for the presence of myocardial fibrosis. Plasma carboxy-terminal propeptide of collagen type I (PICP), carboxy-terminal telopeptide of collagen type I (CITP), and tissue inhibitor of matrix metalloproteinases type I (TIMP-1) were measured as markers of collagen synthesis, degradation, and inhibition of degradation, respectively. Plasma TIMP-1 was significantly elevated in the hypertensive cohort (358 ng/mL versus 253 ng/mL, P<0.001) as were CITP (5.2 microg/L versus 2.9 microg/L, P<0.001), and PICP (200 microg/L versus 166 microg/L, P<0.05). TIMP-1 was significantly elevated in patients with diastolic dysfunction (421 ng/mL versus 283 ng/mL P<0.01) and correlated with markers of diastolic filling, namely E:A ratio (r=0.26, P<0.05) and E Dec (r=0.41, P<0.01). A plasma TIMP-1 level of >500 ng/mL had a specificity of 97% and a positive predictive value of 96% in predicting diastolic dysfunction. In patients with untreated hypertension, there is evidence of increased collagen synthesis, degradation, and inhibition of degradation resulting in fibrosis. Our results demonstrate that plasma TIMP-1 correlates with markers of LV diastolic filling, is predictive of LV dysfunction, and is a potential noninvasive marker of fibrosis.