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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

Images of dot-immunobinding assay of total AGE modified proteins in tears from DR, DNR, and CTL subjects. One dot represented a tear sample from a subject. The left lane was the AGE-BSA standard in various amounts.
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f1: Images of dot-immunobinding assay of total AGE modified proteins in tears from DR, DNR, and CTL subjects. One dot represented a tear sample from a subject. The left lane was the AGE-BSA standard in various amounts.

Mentions: Fifteen tear samples from each group were analyzed by dot-immunobinding assay using anti-AGE antibody. The images of tears from CTL, DNR and DR patients are shown in Figure 1. Based on the dot intensity of the standard AGE modified BSA, the amount of total AGE modified proteins in each tear sample was calculated (Table 2). Compared with the CTL group, tear concentrations of AGE modified proteins were elevated in DR and NDR groups. Concentrations of AGE modified proteins were higher in DR group than DNR group but the difference did not achieve statistical significance (p>0.05). After converting the results to μg AGE modified proteins per mg tear protein, the two DM groups still had higher amount of AGE modified proteins but the statistical difference between CTL and DNR became insignificant, whereas CTL versus DR remained significantly different.


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)

Images of dot-immunobinding assay of total AGE modified proteins in tears from DR, DNR, and CTL subjects. One dot represented a tear sample from a subject. The left lane was the AGE-BSA standard in various amounts.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Images of dot-immunobinding assay of total AGE modified proteins in tears from DR, DNR, and CTL subjects. One dot represented a tear sample from a subject. The left lane was the AGE-BSA standard in various amounts.
Mentions: Fifteen tear samples from each group were analyzed by dot-immunobinding assay using anti-AGE antibody. The images of tears from CTL, DNR and DR patients are shown in Figure 1. Based on the dot intensity of the standard AGE modified BSA, the amount of total AGE modified proteins in each tear sample was calculated (Table 2). Compared with the CTL group, tear concentrations of AGE modified proteins were elevated in DR and NDR groups. Concentrations of AGE modified proteins were higher in DR group than DNR group but the difference did not achieve statistical significance (p>0.05). After converting the results to μg AGE modified proteins per mg tear protein, the two DM groups still had higher amount of AGE modified proteins but the statistical difference between CTL and DNR became insignificant, whereas CTL versus DR remained significantly different.

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