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Tag Archives: Interferon-alpha

Interferon-alpha-induced changes in metallothionein expression: Discussion (Part 3)

IFN-alpha and -beta are well known for inducing MT-II genes in cultured cells , but little is known on expression patterns of MT isoforms in the liver of HCV-infected patients who have received IFN-alpha therapy. To explore changes in hepatic MT isoform expression resulting from the influence of IFN-alpha, we stained for MT-I and MT-II mRNA in HCV-infected patients both pre- and post-therapy. Because MT-II mRNA increased in correspondence with an increase in PCNA labelling index before therapy, MT-II may be involved in cell proliferation in liver that has been damaged by continuous HCV infection. Continue reading

Interferon-alpha-induced changes in metallothionein expression: Discussion (Part 2)

HCV patientsZinc is considered to exert a protective action on liver cell activity and to prevent damage caused by oxidative stress through induction of MT. MT gene expression and protein synthesis are induced by IFN during normal or adequate zinc status, but not during periods of zinc deficiency. As was observed in our previous study, serum zinc levels were decreased in NR patients after completion of IFN therapy, consequently diminishing MT protein in the liver. In addition, the induction of the hepatic MT gene by IFN is so rapid and transient that the occasionally performed liver biopsy probably influences MT levels. Continue reading

Interferon-alpha-induced changes in metallothionein expression: Discussion (Part 1)

The grade of MT staining in the liver was similar in SR and NR patients before IFN-alpha therapy, whereas it was higher in SR than in NR patients after therapy. These data are in agreement with our previous study, suggesting that IFN-alpha-induced hepatic MT may participate in the therapeutic effects of IFN-alpha for HCV patients. In comparison with pretreatment levels of MT, the grade of hepatic MT protein was decreased one year after IFN-alpha therapy. Changes in hepatic MT protein expression after therapy differed according to IFN response; MT was reduced significantly in NR but only slightly in SR. Continue reading

Interferon-alpha-induced changes in metallothionein expression: Results (Part 3)

IFN-alpha therapyThe PCNA index in total patients was decreased significantly after IFN-alpha therapy, but the reduction of PCNA index was larger in SR than in NR patients.

Relationship among MT and HAI score and PCNA labelling index

Before IFN-alpha therapy, the grade of MT-II mRNA staining showed a positive correlation with PCNA labelling index (r=0.58, P<0.05) (Figure 6), but not with HAI scores. Continue reading

Interferon-alpha-induced changes in metallothionein expression: Results (Part 2)

MT mRNA expression in the liver

MT-I and -II mRNA were detected in the cytoplasm of hepa-tocytes. There is a positive correlation between MT-1 and MT-II mRNA expression. Before IFN-alpha therapy, the MT-II mRNA scores were higher than the MT-I mRNA scores. MT-I mRNA scores were reduced after IFN-alpha therapy in both NR and SR patients. MT-II mRNA scores after therapy were reduced in SR patients; however, the scores were unchanged in NR patients. Continue reading

Interferon-alpha-induced changes in metallothionein expression: Results (Part 1)

hepatocytesMT staining in the liver

Immunohistochemical expression of MT was demonstrated in the cytoplasm and nuclei of hepatocytes. The MT staining was clustered around the portal tracts that were enlarged because of inflammatory cells and fibrosis, and focally in the interlobular areas. Serial changes of hepatic MT staining in typical SR and NR patients are shown in Figure 3. The MT in the SR patient was stained markedly in the hepatocytes pre-and post-therapy; however, the MT-positive hepatocytes in the NR patients were diminished post-therapy. The MT staining scores before therapy were similar in the SR and NR patients. Continue reading

Interferon-alpha-induced changes in metallothionein expression: Methods (Part 4)

One pathologist categorized the staining as rare (less than 5% of the cells stained), mild (5% to less than 30%), moderate (30% to less than 60%) or strong (more than 60% of the cells stained) and scored as follows: rare = 1, mild = 2, moderate = 3 or strong = 4. Standards for each grade of MT staining and MT-mRNA expression are shown in Figures 1 and 2.

PCNA labelling index

Liver specimens were allowed to react with the first antibody, a 1:200 dilution of anti-PCNA monoclonal antibody (DAKO Japan Co Ltd, Japan), at 4°C for 12 h, and then with biotin-labelled antirabbit immunoglobulin G antibody and horseradish peroxidase-bound EnVision (DAKO Japan Co Ltd, Japan). Continue reading

Interferon-alpha-induced changes in metallothionein expression: Methods (Part 3)

metallothionein geneAfter the target DNA was denatured, the hybridization mix (50 ng probe per 100 p.L Hybrisol IV) was added. The slides were covered, sealed and stored in a humid chamber at 37°C for 16 h. The slides were washed three times in 0.1 x standard saline citrate at 45°C and then two times for 5 min each in 0.05 M Tris-HCl (pH 7.5). After incubation with a 1:300 dilution of anti-digoxygenin antibody (Roche Diagnostics, Germany) in 0.1% bovine serum albumin/Tris for 1 h at room temperature, the slides were washed again in 0.05 M Tris-HCl (pH 7.5). Continue reading

Interferon-alpha-induced changes in metallothionein expression: Methods (Part 2)

In situ hybridization of MT-I and MT-II mRNA in the liver

To prepare the probes for mRNA, digoxygenin-labelled human MT-I and MT-II complementary DNA probes were prepared by polymerase chain reaction as previously described by Kubota et al , with minor modifications. Briefly, a construct of pUCh MT-I and MT-II as template was amplified by Taq polymerase (TaKaRa taq, Takara Bio Inc, Japan) for 10 to 20 cycles involving denatura-tion at 95°C for 2 min, annealing at 65°C for 1 min and primer extension at 72°C for 1 min. A matrix (Boehringer Mannheim, Germany) containing digoxygenin was added to the last five cycles to label the probe. Continue reading

Interferon-alpha-induced changes in metallothionein expression: Methods (Part 1)

chronic hepatitisHistopathology of chronic hepatitis

Hematoxylin and eosin, Mallory and Periodic Acid-Schiff staining were performed for histological analysis. The Knodell’s histological activity index (HAI) score was assessed before and one year after IFN-alpha therapy. The HAI score was calculated by averaging the scores of three visual fields. Please make sure you read all the information provided above to choose the right medications at the canadian health&care mall to benefit the most from the treatment. At this pharmacy, you will find tons of cheap and efficient meds for any problem.

Immunohistochemical studies of MT in the liver Continue reading

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