Immunochemical Detection of Advanced Glycation End Products in Lens Crystallins From Streptozocin-Induced Diabetic Rat

Nakayama, Hidetaka; Mitsuhashi, Tomoko; Kuwajima, Satoru; Aoki, Satoshi; Kuroda, Yosuke; Itoh, Tomohiro; Nakagawa, Shoichi

doi:10.2337/diab.42.2.345

Cited by 101 publications

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“…AGE may form crosslinks responsible, at least in part, for the dgidity (stiffness) of the vascular wall, and are known to quench the dilatatory effect of the nitric oxide. The low fluorescence intensity of AGE-proteins in hamsters lens crystallins is in agreement with that of rat (22), and may be explained by the occurrence of a reduced number of constitutive proteins in this tissue.…”

Section: Discussionsupporting

confidence: 73%

“…Subsequently, the lipid-free samples were solubilised in alkali as described (22). The extracts were sedimented by 5 min centrifugation at 7428g, and the clear supernatant used for: (i) protein assay, and (ii) measurement of the fluorescence of AGE-adducts.…”

Section: Processing Of Tissuesmentioning

confidence: 99%

“…The mesenteric vascular bed is known to accumulate AGE-collagen during advanced chronological age (19) and in diabetes, which caused also the hypertrophy of the mesenteric vasculature (20,21). The lens crystallin glycated in vitro showed a preferential glycation of oLA crystallin subunits (6); in the diabetic (cataractous) lens crystallin accumulation of AGE-proteins was reported (22,23,24). In vivo treatment with L-arginine had benefic effects in diabetes reducing glomerular basement membrane N-carboxymethyllysine collagen, an AGE generated by "glycoxidation" (25).…”

Section: Introductionmentioning

confidence: 99%

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Age-dependent accumulation of advanced glycation endproducts is accelerated in combined hyperlipidemia and hyperglycemia, a process attenuated by L-arginine

Georgescu¹,

Popov²

2000

AGE

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In this study we have investigated the occurrence of "classical" Amadori rearrangement products of AGEproteins in the vascular mesenteric bed and in the lens of Golden Syrian hamsters (12 weeks old) rendered simultaneous hyperlipidemics-diabetics (HD), or hyperlipidemics (H) for 24 weeks. For the next 4 weeks the hamsters in HD and H groups received by gavage a solution of 3 mM L-arginine, with the intent to look for the potential effects of Larginine on the fluorescence of tissular AGE-proteins. Age-matched normal hamsters were used as controls (C). The AGE-products of proteins, and the AGE-collagen isolated from the mesenteric bed were quantitated by fluorescence spectroscopy at ex: 370 nm/em: 440 nm. The results showed that: (i) compared to the fluorescence levels of AGEproteins detected at C goup, in HD group the fluorescence of AGE-proteins was found 2.78 and 7.41 fold increased in the vascular mesenteric bed and lens, respectively; (ii) in H group the fluorescence of AGEproteins was 2.36 fold augumented in the vascular mesenteric bed, and 5.43 fold in the lens (versus the C goup); (iii) the aging occurring during the 24 weeks of the experiment induced a small increase in AGE-proteins fluorescence in both mesentery (1.76 fold) and lens (3.83 fold), compared to the levels measured in C group at the inception of the study (12 weeks old hamsters); (iv) the fluorescence of AGEproteins in the vascular mesenteric bed and in the lens of hamsters in HD and H groups correlated with the increase in circulating plasma glucose and cholesterol concentrations throughout the experiment; (v) L-arginine dietary supplementation in HD and H groups, diminished the AGE-collagen fluorescence in the mesentery to ~ 35% and ~ 17%, respectively; in the lens the fluorescence of AGE-proteins was reduced to 65-70% of the levels found in HD and H groups (at 24 weeks). This study showed for the first time that simultaneous hyperlipidemia-hyperglyTo whom all correspondence should be addressed: Doina Popov, Ph.D Institute of Cellular Biology and Pathology "Nicolae Simionescu", Bucharest, 8 B.P.Hasdeu Street 79691,Romania telephone: 40 1 411 08 60 fax: 401411 1143 e-mail: popov@ simionescu.instcellbiopath.ro cemia induced an enhanced accumulation of fluorescent AGE-proteins in the mesentery and lens (comparatively to the effect of hyperlipidemia and of chronological aging monitored during the experiment), and that in vivo L-arginine administration decreased the fluorescence of tissular AGE-proteins (AGE-collagen included). The latter observation may bring another area of potential intervention in the adjunct efforts to find out inhibitors of AGE formation, and thus to reduce the increased levels of AGEproteins accumulated in tissues when diabetes is additionally complicated with atherosclerosis.

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Section: Discussionsupporting

confidence: 73%

Section: Processing Of Tissuesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Age-dependent accumulation of advanced glycation endproducts is accelerated in combined hyperlipidemia and hyperglycemia, a process attenuated by L-arginine

Georgescu¹,

Popov²

2000

AGE

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“…Previous studies by several groups have measured AGE levels by immunoassay as well as by specific fluorescence in tissues undergoing diabetic complications [22,23]. The advent of these antibodies to AGEs has allowed immunohistochemical techniques to localise AGE proteins in various sites [24][25][26].…”

Section: Discussionmentioning

confidence: 99%

Advanced glycation end products and their receptors co-localise in rat organs susceptible to diabetic microvascular injury

et al. 1997

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“…Determination of Renal AGEs and TBA-Reactive Substance Levels The renal levels of AGEs were determined by the method of Nakayama et al 24) Minced kidney tissue was delipidated with chloroform and MeOH (2 : 1, v/v) overnight. After washing, the tissue was homogenized with 0.1 M NaOH, followed by centrifugation at 8000 g for 15 min at 4°C.…”

mentioning

confidence: 99%

Protective Effect of Sun Ginseng against Diabetic Renal Damage

Kang

Kim

Yamabe

et al. 2006

Biological & Pharmaceutical Bulletin

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Diabetes mellitus is characterized by hyperglycemia. An abnormally elevated blood glucose level causes oxidative stress and the formation of advanced glycation endproducts (AGEs), which have been closely linked to diabetic complications such as neuropathy, retinopathy and nephropathy. 1,2) In particular, diabetics are at increased risk for several types of kidney disease, and the predominant cause of end stage renal disease in this disorder is diabetic nephropathy. 3,4) However, recent clinical trials suggest that there is no effective treatment for diabetic nephropathy.5) Therefore, preventions of the occurrence and progression of diabetic nephropathy has become a very important issue. Therefore, great effort has been focused on traditional and herbal medicine without toxic effects to find a novel therapeutic agent for diabetic nephropathy.Panax ginseng C. A. MEYER is one of the most widely used herbal medicines in the Orient. It has a wide range of pharmacological and physiological actions, such as antiaging, immunoenhancement, antistress and antitumor. [6][7][8] In addition, several investigations strongly support the evidence that ginseng root possesses anti-diabetic properties, such as inhibition of intestinal glucose absorption, increase in energy expenditure, improving sensitivity to insulin and stimulation of sugar metabolism, etc.9-11) Moreover, ginseng root has been shown in clinical studies to have beneficial effects in diabetic patients. 12,13) Of the two kinds of ginseng, white ginseng (WG) is airdried ginseng, and red ginseng (RG) is produced by steaming raw ginseng at 98-100°C for 2-3 h. RG is reportedly more pharmacologically active than WG.14) These improved biological activities of ginsengs result from changes in the chemical constituents that occur during steam treatment. Ginseng saponins, referred to as ginsenosides, are believed to play a pharmacologically important role. Several investigators have reported new ginsenosides from RG that are not usually found in WG.15) Recently, a method which can enhance the yield of these RG specific components by steaming ginseng at a temperature higher than RG has been developed.16) This heat-processed ginseng, termed sun ginseng (SG), has been reported to have more potent pharmacological activities, such as vasorelaxation, antioxidant and anti-tumor activities.17,18) Furthermore, SG showed stronger peroxynitrite (ONOO Ϫ ) and hydroxyl radical scavenging activities than WG and RG in our previous in vitro study.19) However, the effects of SG on diabetic rats have not been reported yet, and the enhanced radical scavenging activities of SG are thought to be beneficial against diabetic oxidative damage caused by hyperglycemia.Therefore, the protective effect of SG against renal damage caused by oxidative stress or the formation of AGEs under diabetes and its molecular biological mechanism were investigated in this study. MATERIALS AND METHODSReagents Phenylmethylsulfonyl fluoride (PMS) and bactin were purchased from Sigma Chemical Co. (St. Louis, MO, U.S.A.)....

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Immunochemical Detection of Advanced Glycation End Products in Lens Crystallins From Streptozocin-Induced Diabetic Rat

Cited by 101 publications

References 0 publications

Age-dependent accumulation of advanced glycation endproducts is accelerated in combined hyperlipidemia and hyperglycemia, a process attenuated by L-arginine

Age-dependent accumulation of advanced glycation endproducts is accelerated in combined hyperlipidemia and hyperglycemia, a process attenuated by L-arginine

Advanced glycation end products and their receptors co-localise in rat organs susceptible to diabetic microvascular injury

Protective Effect of Sun Ginseng against Diabetic Renal Damage

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