Limits...
Novel biomarkers predict liver fibrosis in hepatitis C patients: alpha 2 macroglobulin, vitamin D binding protein and apolipoprotein AI.

Ho AS, Cheng CC, Lee SC, Liu ML, Lee JY, Wang WM, Wang CC - J. Biomed. Sci. (2010)

Bottom Line: Three serum samples of each fibrotic stage were analyzed by two-dimension difference gel electrophoresis (2D-DIGE).The differential proteins were identified by the cooperation of MALDI-TOF/TOF and MASCOT; then western blotting and Bio-Plex Suspension Array were used to quantify the protein levels.Three prominent candidate biomarkers were identified: alpha 2 macroglobulin (A2M) is up regulated; vitamin D binding protein (VDBP) and apolipoprotein AI (ApoAI) are down regulated.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Gastroenterology, Buddhist Tzu Chi General Hospital, Taipei branch, Taiwan.

ABSTRACT

Background: The gold standard of assessing liver fibrosis is liver biopsy, which is invasive and not without risk. Therefore, searching for noninvasive serologic biomarkers for liver fibrosis is an importantly clinical issue.

Methods: A total of 16 healthy volunteers and 45 patients with chronic hepatitis C virus (HCV) were enrolled (F0: n = 16, F1: n = 7, F2: n = 17, F3: n = 8 and F4: n = 13, according to the METAVIR classification). Three serum samples of each fibrotic stage were analyzed by two-dimension difference gel electrophoresis (2D-DIGE). The differential proteins were identified by the cooperation of MALDI-TOF/TOF and MASCOT; then western blotting and Bio-Plex Suspension Array were used to quantify the protein levels.

Results: Three prominent candidate biomarkers were identified: alpha 2 macroglobulin (A2M) is up regulated; vitamin D binding protein (VDBP) and apolipoprotein AI (ApoAI) are down regulated. The serum concentration of A2M was significantly different among normal, mild (F1/F2) and advanced fibrosis (F3/F4) (p < 0.01). The protein levels of VDBP and ApoAI were significantly higher in normal/mild fibrosis, when compared to those in advanced fibrosis (both p < 0.01).

Conclusions: This study not only reveals three putative biomarkers of liver fibrosis (A2M, VDBP and ApoAI) but also proves the differential expressions of those markers in different stages of fibrosis. We expect that combination of these novel biomarkers could be applied clinically to predict the stage of liver fibrosis without the need of liver biopsy.

Show MeSH

Related in: MedlinePlus

Verification of A2M, VDBP and ApoAI by western blotting (A) and quantification of A2M (B), VDBP (C) and ApoAI (D) by Bio-Plex Suspension Array System. A2M is increased in the F1-F4 stages; VDBP is decreased in the F2-F4 stage and ApoAI is decreased in the F3/F4 stages. **p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2914022&req=5

Figure 3: Verification of A2M, VDBP and ApoAI by western blotting (A) and quantification of A2M (B), VDBP (C) and ApoAI (D) by Bio-Plex Suspension Array System. A2M is increased in the F1-F4 stages; VDBP is decreased in the F2-F4 stage and ApoAI is decreased in the F3/F4 stages. **p < 0.01.

Mentions: Although 2D-DIGE analyses already demonstrated that the protein levels and expressional trends for each candidate biomarkers were apparently distinct in fibrotic stages, using antibody to verify the result was important. We selected two samples of each fibrotic stage to analyze the protein expressions by using western blotting for verifying the protein identification. A2M was detected having higher protein expression in F1-F4 stage than in F0 stage (Fig. 3A). Moreover, VDBP and ApoAI were down regulated (Fig. 3A). Particularly the protein expression of VDBP from mild fibrosis (F0/F1) to advanced fibrosis (F2-F4) was decreased (Fig. 3A and Fig. 3C). The protein expression of ApoAI was changeable only in F3/F4 compared with that in F0-F2 stage (Fig. 3A and Fig. 3D).


Novel biomarkers predict liver fibrosis in hepatitis C patients: alpha 2 macroglobulin, vitamin D binding protein and apolipoprotein AI.

Ho AS, Cheng CC, Lee SC, Liu ML, Lee JY, Wang WM, Wang CC - J. Biomed. Sci. (2010)

Verification of A2M, VDBP and ApoAI by western blotting (A) and quantification of A2M (B), VDBP (C) and ApoAI (D) by Bio-Plex Suspension Array System. A2M is increased in the F1-F4 stages; VDBP is decreased in the F2-F4 stage and ApoAI is decreased in the F3/F4 stages. **p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2914022&req=5

Figure 3: Verification of A2M, VDBP and ApoAI by western blotting (A) and quantification of A2M (B), VDBP (C) and ApoAI (D) by Bio-Plex Suspension Array System. A2M is increased in the F1-F4 stages; VDBP is decreased in the F2-F4 stage and ApoAI is decreased in the F3/F4 stages. **p < 0.01.
Mentions: Although 2D-DIGE analyses already demonstrated that the protein levels and expressional trends for each candidate biomarkers were apparently distinct in fibrotic stages, using antibody to verify the result was important. We selected two samples of each fibrotic stage to analyze the protein expressions by using western blotting for verifying the protein identification. A2M was detected having higher protein expression in F1-F4 stage than in F0 stage (Fig. 3A). Moreover, VDBP and ApoAI were down regulated (Fig. 3A). Particularly the protein expression of VDBP from mild fibrosis (F0/F1) to advanced fibrosis (F2-F4) was decreased (Fig. 3A and Fig. 3C). The protein expression of ApoAI was changeable only in F3/F4 compared with that in F0-F2 stage (Fig. 3A and Fig. 3D).

Bottom Line: Three serum samples of each fibrotic stage were analyzed by two-dimension difference gel electrophoresis (2D-DIGE).The differential proteins were identified by the cooperation of MALDI-TOF/TOF and MASCOT; then western blotting and Bio-Plex Suspension Array were used to quantify the protein levels.Three prominent candidate biomarkers were identified: alpha 2 macroglobulin (A2M) is up regulated; vitamin D binding protein (VDBP) and apolipoprotein AI (ApoAI) are down regulated.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Gastroenterology, Buddhist Tzu Chi General Hospital, Taipei branch, Taiwan.

ABSTRACT

Background: The gold standard of assessing liver fibrosis is liver biopsy, which is invasive and not without risk. Therefore, searching for noninvasive serologic biomarkers for liver fibrosis is an importantly clinical issue.

Methods: A total of 16 healthy volunteers and 45 patients with chronic hepatitis C virus (HCV) were enrolled (F0: n = 16, F1: n = 7, F2: n = 17, F3: n = 8 and F4: n = 13, according to the METAVIR classification). Three serum samples of each fibrotic stage were analyzed by two-dimension difference gel electrophoresis (2D-DIGE). The differential proteins were identified by the cooperation of MALDI-TOF/TOF and MASCOT; then western blotting and Bio-Plex Suspension Array were used to quantify the protein levels.

Results: Three prominent candidate biomarkers were identified: alpha 2 macroglobulin (A2M) is up regulated; vitamin D binding protein (VDBP) and apolipoprotein AI (ApoAI) are down regulated. The serum concentration of A2M was significantly different among normal, mild (F1/F2) and advanced fibrosis (F3/F4) (p < 0.01). The protein levels of VDBP and ApoAI were significantly higher in normal/mild fibrosis, when compared to those in advanced fibrosis (both p < 0.01).

Conclusions: This study not only reveals three putative biomarkers of liver fibrosis (A2M, VDBP and ApoAI) but also proves the differential expressions of those markers in different stages of fibrosis. We expect that combination of these novel biomarkers could be applied clinically to predict the stage of liver fibrosis without the need of liver biopsy.

Show MeSH
Related in: MedlinePlus