Troglitazone Not Only Increases GLUT4 but Also Induces Its Translocation in Rat Adipocytes

Troglitazone Not Only Increases GLUT4 but Also Induces Its Translocation in Rat Adipocytes Mitsuyo Shintani 1 , Haruo Nishimura 2 , Shin Yonemitsu 1 , Yoshihiro Ogawa 1 , Tatsuya Hayashi 1 , Kiminori Hosoda 1 , Gen Inoue 1 and Kazuwa Nakao 1 1 Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, Kyoto, Japan, and the 2 Department of Diabetes and Endocrinology, Osaka Saiseikai Nakatsu Hospital, Osaka, Japan Abstract Thiazolidinediones, insulin-sensitizing agents, have been reported to increase glucose uptake along with the expression of glucose transporters in adipocytes and cardiomyocytes. Recently, we have further suggested that the translocation of GLUT4 is stimulated by thiazolidinediones in L6 myocytes. However, the direct effects of thiazolidinediones on translocation of glucose transporters have not yet been determined. In this study, using hemagglutinin epitope-tagged GLUT4 (GLUT4-HA), we provide direct evidence of the effect of troglitazone on the translocation of GLUT4 in rat epididymal adipocytes. Primary cultures of rat adipocytes were transiently transfected with GLUT4-HA and overexpressed eightfold compared with endogenous GLUT4 in transfected cells. A total of 24 h of treatment with troglitazone (10 −4 mol/l) increased the cell surface level of GLUT4-HA by 1.5 ± 0.03–fold ( P < 0.01) without changing the total amount of GLUT4-HA, whereas it increased the protein level of endogenous GLUT4 (1.4-fold) without changing that of GLUT1. Thus, the direct effect on the translocation can be detected apart from the increase in endogenous GLUT4 content using GLUT4-HA. Troglitazone not only increased the translocation of GLUT4-HA on the cell surface in the basal state but also caused a leftward shift in the dose-response relations between GLUT4-HA translocation and insulin concentration in the medium (ED 50 : from ∼0.1 to 0.03 nmol/l). These effects may partly contribute to the antidiabetic activity of troglitazone in patients with obesity and type 2 diabetes. Footnotes Address correspondence and reprint requests to Haruo Nishimura, MD, PhD, Department of Diabetes and Endocrinology, Osaka Saiseikai Nakatsu Hospital, 2-10-39 Shibata, Kita-ku, Osaka, Japan. E-mail: hnis{at}kuhp.kyoto-u.ac.jp . Received for publication 21 December 2000 and accepted in revised form 27 June 2001. BSA, bovine serum albumin; DMEM, Dulbecco’s modified Eagle’s medium; GLUT4-HA, hemagglutinin epitope-tagged GLUT4; HA, hemagglutinin epitope; KRB