Plasma Pentosidine Levels Measured by a Newly Developed Method Using ELISA in Patients with Chronic Renal Failure

Abstract

The plasma pentosidine levels in patients with renal disease were measured by a simple method which was established for plasma and urinary pentosidine determinations. The method, which can be completed within a few hours, involves pretreating plasma with proteolytic enzyme (pronase) and measuring the concentration of pentosidine in the sample by ELISA using antipentosidine antibodies. The prepared antibodies showed no cross-reaction with the raw materials for pentosidine synthesis or with compounds having similar structures. SDS-PAGE indicated that the antibodies had a high purity. The reaction of the antibodies and keyhole limpet hemocyanin-pentosidine in the competitive ELISA system was inhibited by free pentosidine. Excellent standard curves for pentosidine determination were obtained. In actual measurements of clinical samples from patients, a good correlation (r = 0.9356) was obtained between the values measured by ELISA and HPLC. The plasma pentosidine level in patients with renal disease correlated significantly with plasma creatinine, urea nitrogen, β₂-microglobulin, and creatinine clearance, indicating its usefulness in evaluating the severity of renal disease. A significant elevation in plasma pentosidine levels was observed in mild renal dysfunction, whereas no significant increases in creatinine and urea nitrogen levels were detected, suggesting that the plasma pentosidine level is useful in the early diagnosis of beginning renal failure. In patients with chronic renal failure, no difference in plasma pentosidine levels was observed between diabetic nephropathy and chronic glomerulonephritis, while a significant correlation was observed with phosphatidylcholine hydroperoxide, suggesting the possibility that the plasma pentosidine level reflects injury due to oxidation. From these results, the quantitative measurement method developed by us is judged to be a superior innovation for measuring pentosidine in body fluids. The plasma pentosidine level may be useful for the early diagnosis of mild renal failure and to estimate the degree of the severity of renal diseases.

Relationship between Blood Levels of N-Carboxymethyl-lysine and Pentosidine and the Severity of Microangiopathy in Type 2 Diabetes

“…Takeuchi et al reported that blood CML levels were not associated with HbA 1c in patients with type 2 diabetes [15], while Sanaka et al found that blood pentosidine and blood glucose levels were not correlated [8]. These reports are consistent with our findings, suggesting that the production of CML or pentosidine does not directly reflect the severity of hyperglycemia.…”

“…A significant increase of CML and pentosidine was only noted after the onset of CRF along with the elevation of serum creatinine. Since CML and pentosidine are eliminated by renal excretion, their blood levels are directly proportional to the creatinine level and are inversely proportional to creatinine clearance [7,8]. Therefore, the present results reflected the fact that the blood levels of CML and pentosidine are influenced by renal clearance.…”

“…Height and body weight were also determined for calculation of the BMI. FPG and creatinine were measured by an enzymatic method, HbA 1c was determined by high performance liquid chromatography(HPLC), and CML [7] and pentosidine [8] were measured by enzyme-linked immunosorbent assay (ELISA) as described below. …”

High Serum Level of Pentosidine, an Advanced Glycation End Product (AGE), is a Risk Factor of Patients with Heart Failure

“…This method involved pretreating sera with a proteolytic enzyme (pronase) and then measuring concentrations of pentosidine in the sample by ELISA [13].…”

Ageing in Werner syndrome

“…HC (nmol/mL) was examined by HPLC (39) and pentosidine (μg/mL) was assayed by ELISA as previously described (40). The concentration of MMP-9 to vascular damage by activating MMP-9 (42), and both HC and pentosidine were in general increased with inflammation monitored by hsCRP, both oxidative markers were neither correlated with each other, nor serum levels of hsCRP and MMP-9 in WS.…”