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Advanced glycation end product (AGE) modified proteins in tears of diabetic patients.

Zhao Z, Liu J, Shi B, He S, Yao X, Willcox MD - Mol. Vis. (2010)

Bottom Line: The individual AGE modified proteins were also compared between groups using densitometry.Compared with the CTL group, tear concentrations of AGE modified proteins were significantly elevated in DR and DNR groups.AGE modified proteins in tears may be used as biomarkers to diagnose diabetes and/or diabetic retinopathy.

View Article: PubMed Central - PubMed

Affiliation: Brien Holden Vision Institute, Sydney, NSW 2052, Australia. z.zhao@brienholdenvision.org

ABSTRACT

Purpose: High glucose level in diabetic patients may lead to advanced glycation end product (AGE) modified proteins. This study investigated AGE modified proteins in tears and compared their levels in diabetic patients (DM) with non-diabetic controls (CTL).

Methods: Basal tears were collected from DM with (DR) or without (DNR) retinopathy and CTL. Total AGE modified proteins were detected quantitatively by a dot immunobinding assay. The AGE modified proteins were separated in 1D- and 2D-SDS gels and detected by western-blotting. The individual AGE modified proteins were also compared between groups using densitometry.

Results: Compared with the CTL group, tear concentrations of AGE modified proteins were significantly elevated in DR and DNR groups. The concentration of AGE modified proteins in diabetic tears were positively correlated with AGE modified hemoglobin (HbA1c) and postprandial blood glucose level (PBG). Western blotting of AGE modified proteins from 1D-SDS gels showed several bands, the major one at around 60 kDa. The intensities of AGE modified protein bands were higher in DM tears than in CTL tears. Western blotting from 2D-SDS gels showed a strongly stained horizontal strip, which corresponded to the major band in 1D-SDS gels. Most of the other AGE modified protein species were within molecular weight of 30-60 kDa, PI 5.2-7.0. Densitometry analysis demonstrated several AGE modified proteins were elevated in DR or DNR tears.

Conclusions: Total and some individual AGE modified proteins were elevated in DM tears. AGE modified proteins in tears may be used as biomarkers to diagnose diabetes and/or diabetic retinopathy.

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Related in: MedlinePlus

2D western blotting images of AGE modified tear proteins from DR, DNR, and CTL samples and a Sypro ruby stained 2D gel image of a tear sample.
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f4: 2D western blotting images of AGE modified tear proteins from DR, DNR, and CTL samples and a Sypro ruby stained 2D gel image of a tear sample.

Mentions: The western blot of the 2D gels showed many spots in every sample, indicating that many protein species were potentially AGE modified, even in CTL tears (Figure 4). Compared to CTL tears, DM tears (DR and DNR) again were demonstrated to contain more AGE modified protein species. The locations of the AGE modified protein spots were very different from the location of tear protein spots in 2D gels stained with Sypro Ruby, indicating that most of the glycated proteins were of low abundance and not detectable by Sypro Ruby staining. A long strongly stained horizontal strip, which corresponded to the major band in 1D gels, at MW of approximately 60 kDa and PI of 4.5–6.2 appeared in all samples. Most of the other AGE modified protein species were within a range of molecular weight 30–60 kDa, PI 5.2–7.0 (Figure 4). The patterns of AGE modified protein staining were similar in all the tear samples but varied in intensity.


Advanced glycation end product (AGE) modified proteins in tears of diabetic patients.

Zhao Z, Liu J, Shi B, He S, Yao X, Willcox MD - Mol. Vis. (2010)

2D western blotting images of AGE modified tear proteins from DR, DNR, and CTL samples and a Sypro ruby stained 2D gel image of a tear sample.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2927418&req=5

f4: 2D western blotting images of AGE modified tear proteins from DR, DNR, and CTL samples and a Sypro ruby stained 2D gel image of a tear sample.
Mentions: The western blot of the 2D gels showed many spots in every sample, indicating that many protein species were potentially AGE modified, even in CTL tears (Figure 4). Compared to CTL tears, DM tears (DR and DNR) again were demonstrated to contain more AGE modified protein species. The locations of the AGE modified protein spots were very different from the location of tear protein spots in 2D gels stained with Sypro Ruby, indicating that most of the glycated proteins were of low abundance and not detectable by Sypro Ruby staining. A long strongly stained horizontal strip, which corresponded to the major band in 1D gels, at MW of approximately 60 kDa and PI of 4.5–6.2 appeared in all samples. Most of the other AGE modified protein species were within a range of molecular weight 30–60 kDa, PI 5.2–7.0 (Figure 4). The patterns of AGE modified protein staining were similar in all the tear samples but varied in intensity.

Bottom Line: The individual AGE modified proteins were also compared between groups using densitometry.Compared with the CTL group, tear concentrations of AGE modified proteins were significantly elevated in DR and DNR groups.AGE modified proteins in tears may be used as biomarkers to diagnose diabetes and/or diabetic retinopathy.

View Article: PubMed Central - PubMed

Affiliation: Brien Holden Vision Institute, Sydney, NSW 2052, Australia. z.zhao@brienholdenvision.org

ABSTRACT

Purpose: High glucose level in diabetic patients may lead to advanced glycation end product (AGE) modified proteins. This study investigated AGE modified proteins in tears and compared their levels in diabetic patients (DM) with non-diabetic controls (CTL).

Methods: Basal tears were collected from DM with (DR) or without (DNR) retinopathy and CTL. Total AGE modified proteins were detected quantitatively by a dot immunobinding assay. The AGE modified proteins were separated in 1D- and 2D-SDS gels and detected by western-blotting. The individual AGE modified proteins were also compared between groups using densitometry.

Results: Compared with the CTL group, tear concentrations of AGE modified proteins were significantly elevated in DR and DNR groups. The concentration of AGE modified proteins in diabetic tears were positively correlated with AGE modified hemoglobin (HbA1c) and postprandial blood glucose level (PBG). Western blotting of AGE modified proteins from 1D-SDS gels showed several bands, the major one at around 60 kDa. The intensities of AGE modified protein bands were higher in DM tears than in CTL tears. Western blotting from 2D-SDS gels showed a strongly stained horizontal strip, which corresponded to the major band in 1D-SDS gels. Most of the other AGE modified protein species were within molecular weight of 30-60 kDa, PI 5.2-7.0. Densitometry analysis demonstrated several AGE modified proteins were elevated in DR or DNR tears.

Conclusions: Total and some individual AGE modified proteins were elevated in DM tears. AGE modified proteins in tears may be used as biomarkers to diagnose diabetes and/or diabetic retinopathy.

Show MeSH
Related in: MedlinePlus