Effect of aldose reductase inhibitor (Ponalrestat) on erythrocyte Na,K-ATPase activity in non-insulin-dependent diabetic patients with polyneuropathy.
The binding capacity of ouabain to erythrocyte Na,K-ATPase was determined to analyze alterations in this enzyme activity in non-insulin-dependent diabetic patients. A significant (p less than 0.001) reduction of the binding capacity of ouabain was found in erythrocytes obtained from the diabetic patients with polyneuropathy (0.51 +/- 0.02 pmol/10(9) erythrocytes, m +/- SE, n = 14) as compared with the patients without neuropathy (0.67 +/- 0.02, n = 14) or age-matched control subjects (0.71 +/- 0.04, n = 11). Accordingly, the effect of an aldose reductase inhibitor (ARI; Ponalrestat) on erythrocyte Na,K-ATPase activity was studied following two or three months oral administration in seven of the diabetic patients with polyneuropathy. After treatment with Ponalrestat the mean binding capacity of ouabain was significantly increased from 0.53 +/- 0.04 to 0.57 +/- 0.03 (p less than 0.05 by paired t-test). Furthermore, enzyme kinetics showed that in normal subjects the apparent Km and Vmax of erythrocyte membrane Na,K-ATPase were 0.51 +/- 0.07 mM (n = 5, m +/- SE) and 7.19 +/- 0.27 nmol Pi/mg protein/min (n = 5, m +/- SE), respectively. The Vmax with 3 mM ATP was significantly (p less than 0.05) decreased in the diabetic patients with polyneuropathy as compared with age-matched control subjects. However, the apparent Km did not change. Finally, the in vitro effect of Ponalrestat was examined in erythrocyte membrane fractions from the diabetic patients with polyneuropathy. The activity of erythrocyte membrane Na,K-ATPase was found to be directly stimulated about 1.2 fold by the addition of pharmacological doses of Ponalrestat (10(-10), 10(-8), 10(-6) M).(ABSTRACT TRUNCATED AT 250 WORDS)
Umeda
F
F
Third Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Noda
K
K
Hashimoto
T
T
Yamashita
T
T
Nawata
H
H
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors*; D003924:Diabetes Mellitus, Type 2 / Q000097:blood / Q000188:drug therapy / Q000201:enzymology*; D003929:Diabetic Neuropathies / Q000097:blood / Q000201:enzymology* / Q000503:physiopathology; D004910:Erythrocyte Membrane / Q000201:enzymology*; D005260:Female; D006801:Humans; D007004:Hypoglycemic Agents / Q000627:therapeutic use*; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D010042:Ouabain / Q000378:metabolism; D010793:Phthalazines / Q000627:therapeutic use*; D011485:Protein Binding; D011724:Pyridazines / Q000627:therapeutic use*; D012016:Reference Values; D000254:Sodium-Potassium-Exchanging ATPase / Q000097:blood*; D013401:Sugar Alcohol Dehydrogenases / Q000037:antagonists & inhibitors*
D007004:Hypoglycemic Agents; D010793:Phthalazines; D011724:Pyridazines; C042829:ponalrestat; D010042:Ouabain; D013401:Sugar Alcohol Dehydrogenases; D000449:Aldehyde Reductase; D000254:Sodium-Potassium-Exchanging ATPase