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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

Image analysis to detect expression differences of the individual AGE modified protein spot between the groups. Only the spots showing significant differences were included in the figure table. No significant differences were detected between DNR and DR groups.
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f5: Image analysis to detect expression differences of the individual AGE modified protein spot between the groups. Only the spots showing significant differences were included in the figure table. No significant differences were detected between DNR and DR groups.

Mentions: Image analysis showed that many spots were upregulated in DR tears or DNR tears when compared with CTL tears (Figure 5). This was most obvious for spot 18. Taking CTL tears as a reference, its intensity increased 7.4 fold and 3.1 fold in DR and DNR tears, respectively. The intensity of spot 5 increased 4.1 fold in DR tears but did not increase in DNR tears. Conversely, the intensity of spot 35 was elevated 2.9 fold in DNR tears but did not change in DR tears. No significant differences were detected for spot intensities between DNR group and DR group for any of the spots.


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)

Image analysis to detect expression differences of the individual AGE modified protein spot between the groups. Only the spots showing significant differences were included in the figure table. No significant differences were detected between DNR and DR groups.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: Image analysis to detect expression differences of the individual AGE modified protein spot between the groups. Only the spots showing significant differences were included in the figure table. No significant differences were detected between DNR and DR groups.
Mentions: Image analysis showed that many spots were upregulated in DR tears or DNR tears when compared with CTL tears (Figure 5). This was most obvious for spot 18. Taking CTL tears as a reference, its intensity increased 7.4 fold and 3.1 fold in DR and DNR tears, respectively. The intensity of spot 5 increased 4.1 fold in DR tears but did not increase in DNR tears. Conversely, the intensity of spot 35 was elevated 2.9 fold in DNR tears but did not change in DR tears. No significant differences were detected for spot intensities between DNR group and DR group for any of the spots.

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