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J. Biol. Chem., Vol. 262, Issue 1, 189-197, Jan, 1987
WT Garvey, JM Olefsky, S Matthaei and S Marshall
We have previously shown in primary cultured rat adipocytes that insulin
acts at receptor and multiple postreceptor sites to decrease insulin's
subsequent ability to stimulate glucose transport. To examine whether
D-glucose can regulate glucose transport activity and whether it has a role
in insulin-induced insulin resistance, we cultured cells for 24 h in the
absence and presence of various glucose and insulin concentrations. After
washing cells and allowing the glucose transport system to deactivate, we
measured basal and maximally insulin- stimulated 2-deoxyglucose uptake
rates (37 degrees C) and cell surface insulin binding (16 degrees C).
Alone, incubation with D-glucose had no effect on basal or maximal glucose
transport activity, and incubation with insulin, in the absence of glucose,
decreased maximal (but not basal) glucose transport rates only 18% at the
highest preincubation concentration (50 ng/ml). However, in combination,
D-glucose (1-20 mM) markedly enhanced the long-term ability of insulin
(1-50 ng/ml) to decrease glucose transport rates in a dose-responsive
manner. For example, at 50 ng/ml preincubation insulin concentration, the
maximal glucose transport rate fell from 18 to 63%, and the basal uptake
rate fell by 89%, as the preincubation D-glucose level was increased from 0
to 20 mM. Moreover, D-glucose more effectively promoted decreases in basal
glucose uptake (Ki = 2.2 +/- 0.4 mM) compared with maximal transport rates
(Ki = 4.1 +/- 0.4 mM) at all preincubation insulin concentrations (1-50
ng/ml). Similar results were obtained when initial rates of
3-O-methylglucose uptake were used to measure glucose transport. D-glucose,
in contrast, did not influence insulin-induced receptor loss. In other
studies, D-mannose and D-glucosamine could substitute for D-glucose to
promote the insulin-induced changes in glucose transport, but other
substrates such as L-glucose, L-arabinase, D-fructose, pyruvate, and
maltose were without effect. Also, non- metabolized substrates which
competitively inhibit D-glucose uptake (3- O-methylglucose, cytochalasin B)
blocked the D-glucose plus insulin effect.(ABSTRACT TRUNCATED AT 400 WORDS)
Glucose and insulin co-regulate the glucose transport system in primary cultured adipocytes. A new mechanism of insulin resistance
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