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Peroxiredoxin 2: a potential biomarker for early diagnosis of hepatitis B virus related liver fibrosis identified by proteomic analysis of the plasma.

Lu Y, Liu J, Lin C, Wang H, Jiang Y, Wang J, Yang P, He F - BMC Gastroenterol (2010)

Bottom Line: The alteration was further confirmed by western blotting.Four variables (PT, Pre, HA and Prx II) were selected from the 25 variables to construct the decision tree.This study showed that 2-D DIGE-based proteomic analysis of the plasma was helpful in screening for new plasma biomarkers for liver disease.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Chemistry, Fudan University, Shanghai, China.

ABSTRACT

Background: Liver fibrosis is a middle stage in the course of chronic Hepatitis B virus (HBV) infection, which will develop into cirrhosis and eventually hepatocellular carcinoma (HCC) if not treated at the early stage. Considering the limitations and patients' reluctance to undergo liver biopsy, a reliable, noninvasive diagnostic system to predict and assess treatment and prognosis of liver fibrosis is needed. The aim of this study was to identify biomarkers for early diagnosis of HBV related liver fibrosis.

Method: Plasma samples from 7 healthy volunteers and 27 HBV infected patients with different stages of fibrosis were selected for 2-DIGE proteomic screening. One-way ANOVA analysis was used to assess differences in protein expression among all groups. The alteration was further confirmed by western blotting. Plasma levels of 25 serological variables in 42 healthy volunteers and 68 patients were measured to establish a decision tree for the detection of various stages fibrosis.

Result: The up-regulated proteins along with fibrosis progress included fibrinogen, collagen, macroglobulin, hemopexin, antitrypsin, prealbumin and thioredoxin peroxidase. The down-regulated proteins included haptoglobin, serotransferrin, CD5 antigen like protein, clusterin, apolipoprotein and leucine-rich alpha-2-glycoprotein. For the discrimination of milder stage fibrosis, the area under curve for Prx II was the highest. Four variables (PT, Pre, HA and Prx II) were selected from the 25 variables to construct the decision tree. In a training group, the correct prediction percentage for normal control, milder fibrosis, significant fibrosis and early cirrhosis was 100%, 88.9%, 95.2% and 100%, respectively, with an overall correct percent of 95.9%.

Conclusion: This study showed that 2-D DIGE-based proteomic analysis of the plasma was helpful in screening for new plasma biomarkers for liver disease. The significant up-expression of Prx II could be used in the early diagnosis of HBV related liver fibrosis.

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Related in: MedlinePlus

Protein expressions of Prx II and CLU in plasma by Western blot analysis. A: Single samples of normal and liver fibrosis. Immunoblotting with Prx II or CLU polyclonal antibody following SDS-PAGE was performed as described in Section 2.7. The films were scanned and the OD of each band in the film was evaluated by QuantitiOne software. B: The change of Prx II expression is similar to DIGE result (Figure 2.B). C: The change of CLU expression is similar to DIGE result (Figure 3.B).
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Figure 4: Protein expressions of Prx II and CLU in plasma by Western blot analysis. A: Single samples of normal and liver fibrosis. Immunoblotting with Prx II or CLU polyclonal antibody following SDS-PAGE was performed as described in Section 2.7. The films were scanned and the OD of each band in the film was evaluated by QuantitiOne software. B: The change of Prx II expression is similar to DIGE result (Figure 2.B). C: The change of CLU expression is similar to DIGE result (Figure 3.B).

Mentions: To confirm the differential expression of Prx II, western blotting analysis was also performed using polyclonal antibodies against Prx II (Figure 4A). Because of the huge diversity of protein concentration in plasma among individuals, we did not normalize the result of western blot with a house-keeping protein as usual. Instead, we analyzed the expression of CLU from the same lane of each sample to avoid errors in sample loading and membrane transferring. As expected, the changes of both proteins were similar to that of DIGE result (Figure 4B,C), Compared to normal plasma, Prx II showed to be highly present in all stage of fibrosis plasma, although the up-regulation of Prx II was withdrawn at S3 stage. The presence of CLU showed to decrease continuously with the progress of fibrosis. Thus, the up-expression of Prx II was reliable.


Peroxiredoxin 2: a potential biomarker for early diagnosis of hepatitis B virus related liver fibrosis identified by proteomic analysis of the plasma.

Lu Y, Liu J, Lin C, Wang H, Jiang Y, Wang J, Yang P, He F - BMC Gastroenterol (2010)

Protein expressions of Prx II and CLU in plasma by Western blot analysis. A: Single samples of normal and liver fibrosis. Immunoblotting with Prx II or CLU polyclonal antibody following SDS-PAGE was performed as described in Section 2.7. The films were scanned and the OD of each band in the film was evaluated by QuantitiOne software. B: The change of Prx II expression is similar to DIGE result (Figure 2.B). C: The change of CLU expression is similar to DIGE result (Figure 3.B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2959091&req=5

Figure 4: Protein expressions of Prx II and CLU in plasma by Western blot analysis. A: Single samples of normal and liver fibrosis. Immunoblotting with Prx II or CLU polyclonal antibody following SDS-PAGE was performed as described in Section 2.7. The films were scanned and the OD of each band in the film was evaluated by QuantitiOne software. B: The change of Prx II expression is similar to DIGE result (Figure 2.B). C: The change of CLU expression is similar to DIGE result (Figure 3.B).
Mentions: To confirm the differential expression of Prx II, western blotting analysis was also performed using polyclonal antibodies against Prx II (Figure 4A). Because of the huge diversity of protein concentration in plasma among individuals, we did not normalize the result of western blot with a house-keeping protein as usual. Instead, we analyzed the expression of CLU from the same lane of each sample to avoid errors in sample loading and membrane transferring. As expected, the changes of both proteins were similar to that of DIGE result (Figure 4B,C), Compared to normal plasma, Prx II showed to be highly present in all stage of fibrosis plasma, although the up-regulation of Prx II was withdrawn at S3 stage. The presence of CLU showed to decrease continuously with the progress of fibrosis. Thus, the up-expression of Prx II was reliable.

Bottom Line: The alteration was further confirmed by western blotting.Four variables (PT, Pre, HA and Prx II) were selected from the 25 variables to construct the decision tree.This study showed that 2-D DIGE-based proteomic analysis of the plasma was helpful in screening for new plasma biomarkers for liver disease.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Chemistry, Fudan University, Shanghai, China.

ABSTRACT

Background: Liver fibrosis is a middle stage in the course of chronic Hepatitis B virus (HBV) infection, which will develop into cirrhosis and eventually hepatocellular carcinoma (HCC) if not treated at the early stage. Considering the limitations and patients' reluctance to undergo liver biopsy, a reliable, noninvasive diagnostic system to predict and assess treatment and prognosis of liver fibrosis is needed. The aim of this study was to identify biomarkers for early diagnosis of HBV related liver fibrosis.

Method: Plasma samples from 7 healthy volunteers and 27 HBV infected patients with different stages of fibrosis were selected for 2-DIGE proteomic screening. One-way ANOVA analysis was used to assess differences in protein expression among all groups. The alteration was further confirmed by western blotting. Plasma levels of 25 serological variables in 42 healthy volunteers and 68 patients were measured to establish a decision tree for the detection of various stages fibrosis.

Result: The up-regulated proteins along with fibrosis progress included fibrinogen, collagen, macroglobulin, hemopexin, antitrypsin, prealbumin and thioredoxin peroxidase. The down-regulated proteins included haptoglobin, serotransferrin, CD5 antigen like protein, clusterin, apolipoprotein and leucine-rich alpha-2-glycoprotein. For the discrimination of milder stage fibrosis, the area under curve for Prx II was the highest. Four variables (PT, Pre, HA and Prx II) were selected from the 25 variables to construct the decision tree. In a training group, the correct prediction percentage for normal control, milder fibrosis, significant fibrosis and early cirrhosis was 100%, 88.9%, 95.2% and 100%, respectively, with an overall correct percent of 95.9%.

Conclusion: This study showed that 2-D DIGE-based proteomic analysis of the plasma was helpful in screening for new plasma biomarkers for liver disease. The significant up-expression of Prx II could be used in the early diagnosis of HBV related liver fibrosis.

Show MeSH
Related in: MedlinePlus