Glycation and glycoxidation studies in vitro on isolated human vitreous collagen.
BACKGROUND
Structural and functional impairment in vitreous collagen plays an important role in the pathogenesis of diabetic retinopathy. Collagen being a long-lived protein is prone to both glycation and glycoxidation, resulting in accumulation of advanced glycation end products (AGE). The objective of our study was to explore the extent of glycation by glucose, and iron- and copper-mediated glycoxidation of human vitreous collagen, and also to study the beneficial effects of lysine, inositol and aminoguanidine as antiglycating and anti-cross linking agents.


METHODS
Vitreous from human donor eyeballs was pooled and collagen was extracted using 0.9 M NaCl. Collagen was estimated by measuring the hydroxyproline content. The extracted collagen was used for glycation and glycoxidation studies. Glycation studies were conducted using U14C glucose, along with anti-glycating agents, such as lysine and aminoguanidine. Metal-mediated glycoxidation studies were done by measuring collagen content in cyanogen bromide insoluble fraction, in the presence and absence of an anti-cross linking agent, inositol.


RESULTS
Human vitreous collagen extractable with 0.9 M NaCl was glycated by glucose at 5 and 10 mM concentrations under physiological conditions of temperature and pH. An anti-glycating effect was exhibited by lysine, inositol and aminoguanidine, of which lysine was the best (76% antiglycating activity) followed by inositol. Inositol was also found to be useful in inhibiting glycoxidation.


CONCLUSIONS
Vitreous collagen undergoes glycation, as well as iron- and copper-mediated glycoxidation, leading to possible structural and functional impairment. Glycation and glycoxidation are inhibited, significantly by lysine and inositol respectively.
Sulochana
Konerirajapuram N
KN
Biochemistry Research Department, Vision Research Foundation, Sankara Nethralaya, Chennai, India.
Ramprasad
Srinivasan
S
Coral
Karnuakaran
K
Lakshmi
Soundarajan
S
Punitham
Ranganathan
R
Narayanasamy
Angayarkanni
A
Ramakrishnan
Sivaramakrishna
S
D000818:Animals; D002417:Cattle; D003094:Collagen / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D006909:Hydroxyproline / Q000032:analysis; D010084:Oxidation-Reduction; D014822:Vitreous Body / Q000737:chemistry*
D003094:Collagen; D005947:Glucose; D006909:Hydroxyproline