Archives of the Balkan Medical Union

Copyright 2014 CELSIUS

vol. 49, no. 3, pp. 275-279

September 2014

ORIGINAL PAPER

THE FUNCTION OF INSULIN RECEPTOR (IRF) IN EVOLUTION

OF OBESITY, CORRELATION WITH ADIPOCYTES DIAMETERS
ELENA POPA, AGNES BACUCA, ALAMIR DIAA, MARIA MAIDANIUC, RODICA PETROVANU,
ADORATA ELENA COMAN
Department of Primary Care, School of Medicine, University of Medicine and Pharmacy of Iai, Romania

S UMMARY
In molecular biology, the insulin receptor is a membrane receptor
that is activated by insulin. It belongs to the large class of tyrosine
kinase receptors. In evolution of obesity, insulin-resistance
development depends on insulin receptor function (IRF) (insulin
receptor function, affinity, number). The study group consists of
45 obese patients without co -morbidity and a control group, 9
cases, non-obese (a case-control design of study). The data picked
up and correlated were: radioactivity of R, age of obesity, DAM
(medium diameters of adipocytes), insulinaemia (fasting and 1h
on OGTT). The measurement of IRI shows a negative association, the sub-group with low IRF has high level of fasting IRI,
p<0,01, and 1h, p<0,005. The insulin receptor function is
correlated with fasting insulinaemia and 1h after OGTT, for all
sub-group of study. The main characteristics of sub-groups is grade
of obesity quantified by BMI (body mass index). Comparing BMI
with DAM we obtained a direct correlation, the highest BMI the
largest DAM. There is a negative correlation between DAM and
IRF, the largest DAM the lowest affinity of insulin receptor.
Abreviation: the function of insulin receptor (IRF), radioactivity
of insulin receptor (IRI), insulin receptor (IR), medium diameters
of adipocytes (DAM), oral glucose tolerance test (OGTT).
Key words: insulinemia, insulin receptor, obesity, adipocytes
diametres

I NTRODUCTION
n molecular biology, the insulin receptor is a
membrane receptor that is activated by insulin. It
belongs to the large class of tyrosine kinase
receptors (1).

Correspondence address:

R SUM
La fonction du rcepteur dinsuline (FRI) dans lvolution
de lobsit, la corrlation avec les diamtres adipocytaires
Dans la biologie molculaire, le rcepteur dinsuline est un rcepteur de surface, activ par linsuline. Il appartient la grande
classe des rcepteurs de la tyrosine-kynase. Dans lvolution de
lobsit le dveloppement de la rsistance linsuline dpend de
la fonction du rcepteur dinsuline (FRI) (affinit, nombre,
fonction). Le groupe dtude est form de 45 patients obses sans
morbidits associes et le groupe de contrle, de 9 cas, patients
non-obses. Ltude est une analyse de type cas-contrle. Les
donnes ramasses et analyses sont: la radioactivit du rcepteur
dinsuline (RRI), lge de lobsit, le DMA (le diamtre moyen
de ladipocyte), linsulinmie ( jeun et aprs lpreuve orale de
tolrance au glucose, lOGTT). Les valeurs de la RRI montrent
une association ngative: le sous-groupe de la FRI basse a un
niveau lev de RRI jeun (p<0,01) et aprs OGTT (p<0,005).
La fonction du rcepteur dinsuline est corrle ainsi aux niveaux
de linsulinmie, jeun et aprs 1 heure (OGTT) pour tous les
sous-groupes tudis. Les traits caractristiques principaux des
sous groupes sont le degr de lobsit quantifi par lindice de
masse corporelle (IMC). Au contraire,en comparant lIMC avec le
DMA, nous avons obtenu une corrlation directe: tant lev
lIMC, que large le DMA. Entre le DMA et la FRI il y a une
corrlation inverse:tant grand le DMA que faible la FRI.
Mots clefs: insulinmie, rcepteur dinsuline, obsit,
diamtres adipocytaires

Two alpha subunits and two beta subunits make

together the insulin receptor. The beta subunits pass through
the cellular membrane and are linked by disulfide bonds. The
alpha and beta subunits are encoded by a single gene (INSR).
The insulin receptor has also recently been designated
CD220 (cluster of differentiation 220)(fig. 1)(1,2).

Adorata Elena Coman, Lecturer, MD, PhD

Department of Primary Medicine, School of Medicine
Gr. T. Popa University of Medicine and Pharmacy, 16 University Street, 700115, Iai, Romania
Phone/Fax +40232261195,
e-mail:acoman@iasi.mednet.ro

THE FUNCTION OF INSULIN RECEPTOR (IRF) IN EVOLUTION OF OBESITY ... - POPA et al

vol. 49, no. 3, 276

Figure 2 - Effect of insulin on glucose uptake and metabolism

(6)
Figure 1 - Insulin receptor structure (Wikipedia) (1)

Insulin binds to its receptor which in turn starts many

protein activation cascades (2,3). These include: translocation of Glut-4 transporter to the plasma membrane and
influx of glucose, glycogen synthesis, glycolysis and fatty
acid synthesis (3,4,5,6)(Fig. 2).
In evolution of obesity, insulin-resistance development
depends on insulin receptor function (IRF) (insulin receptor
function, affinity, number). Insulin-resistance and hyperinsulinaemia appearance implies a lot of structures and
systems (7). That is why the statistical calculation, HOMA-IR
(Homeostasis Assessement-InsulinResistance) and HOMA-IS
(Homeostasis Assessement-InsulinSensibility), together with
euglycemic-hyperinsulinemic clamp are the gold standard.
More, there are a lot of experimental data which give the
evidence of direct implication of IRF. In fact, IRF measures
the amplitude of Insulin captation on the level of target
insulin-dependent tissues (7,8,9).
The insulin-receptor inter-reaction is the trigger for a
complex chain production peptides involving membrane
cells, cytoplasm and mitochondria structures, and nucleus.
Even so the importance of IRF remains. The basic
methodology to quantify IRF consists of marking insulin
receptor (IR) with 125I and measuring the quantity of 125I
captation (11,12).

M ATERIAL

AND

M ETHOD

The study group consists of 45 obese patients without

co -morbidity and a control group, 9 cases, non-obese (a
case-control design of study). The study of IRF was made
on adipose tissue obtained by peri-umbilical biopsy on
obese and normal subjects, during surgical intervention.
For all patients we obtained an informed acceptation.
We also measured insulinaemia during OGTT (oral
glucose tolerance test), and as morphological parameters
medium diameters of adipocytes (DAM).

The technique of quantification of IRF

Adipocytes were isolated using collagenase. The
adipocytes deposits were suspended on HEPS solution and
treated with 125I- insulin (ROTOP). The radioactivity of

deposits was measured with Gamma-Mrek counter. As

reference we consider 100% activity, only 125I-insulin
activity, without adipocytes.
There were 5 measurements by considering a medium
value, for each sample. The radioactivity measured is
equivalent with IRF reported to 100% activity, as control
sample. The stain used was Oil Reed, a specific stain for
adipose tissue. After coloration the material is put on
cryostat, sectioned with microtome, 15 micrometer per
section. The adipocytes are well defined and the coloration
is yellow-reddish (for Oil Reed stain).

The technique of measurement of DA

(diameter of adipocytes)
The measurement of DA (diameter of adipocytes) was
made using an ocular micrometer (x10) with immersion
(x100), standard set on a micrometer (m) scale, using a
Brker-Trck chamber. This has in center a secondary scale
that divided one more the scale in chambers of 5 m length
(Thoma chamber). DAs were measured to 300 cells from
each patient (10 fields with 30 cells) and we quantified
DAM (medium adipocytes diameter) with SD (standard
deviation).
We calculated to each group age of obesity, BMI,
insulinaemia (IRI) (fasting IRI and 1h on OGTT) as well
as glycemia (medium values and SD)(Table 1).
The data analysis of BMI (body mass index) reported 3
sub-groups of obesity:
Group I, BMI=427,5 kg/mp,
Group II, BMI= 367,7 kg/pm,
Group III, BMI= 293,3 kg/mp. The three subgroup of study were selected by BMI; for each group
the selection included all decades of age and both
genders.

R ESULTS
All data was analyzed compared with control group and,
also between sub-groups. The main characteristic of subgroups is made by BMI, different grade of obesity, merely the
same medium age of studied population. The data picked up
and correlated was: radioactivity (function) of insulin R

Archives of the Balkan Medical Union

September 2014, 277

Table 1 - Characteristics
of three groups of study

(IRF), age of obesity, DAM (medium diameters of

adipocytes), insulinaemia (fasting and 1h on OGTT).
According to table, the measurement of RF shows an
inversely correlation between RF and BMI: the highest subgroup of BMI (427,5 kg/mp) has the lowest RF
activity (12,40, 66,93%)(table 1). This report is maintained
to all three sub-groups comparing with normal: group I
(66,93%), group II (67,84%), group III (87,31%) and normal
group (100%). Even for the third group with BMI of
293,3kg/mp there are significant differences with control
group (16,74 vs 18,6).
The measurement of IRI shows a negative association,
the sub-group with low IRF has high level of fasting IRI,
p<0,01, and 1h, p<0,005 (fig. 3).
As we compared the sub-group of study, we obtain a

Figure 3- Correlation
between IRF and IRI
a jeun and 1 h

Figure 4 - Medium IRF

on groups of study

negative correlation between IRF and DAM as main

characteristics: for larger adipocytes lower activity of
insulin receptor (fig. 4). It is important to mention that
DAM is a statistically medium value and the variations of
adipocytes sizes are very large.
The regression curves show the negative correlation
between function of insulin receptor, IRF and medium size
of adipocytes. For IRF (%) correlated with DAM, p<0,05
(fig. 5).

D ISCUSSION
Tyrosine kinase receptors, including the insulin receptor,
mediate their activity by causing the addition of a
phosphate group to particular tyrosines on certain proteins

THE FUNCTION OF INSULIN RECEPTOR (IRF) IN EVOLUTION OF OBESITY ... - POPA et al

vol. 49, no. 3, 278

Figure 5 - Correlation
between IRF and DAM
on study groups

within a cell. The "substrate" proteins which are phosphorylated by the Insulin Receptor include a protein called
"IRS-1" for "insulin receptor substrate 1". IRS-1 binding and
phosphorylation eventually leads to an increase in the high
affinity glucose transporter (Glut4) molecules on the outer
membrane of insulin-responsive tissues, including muscle
cells and adipose tissue, and therefore to an increase in the
uptake of glucose from blood into these tissues(13,14).
Briefly, the glucose transporter (Glut4) is transported
from cellular vesicles to the cell surface, where it then can
mediate the transport of glucose into the cell. The main
activity of activation of the insulin receptor is inducing
glucose uptake. For this reason "insulin insensitivity", or a
decrease in insulin receptor signaling, leads to diabetes
mellitus type 2 - the cells are unable to take up glucose,
and the result is hyperglycemia (an increase in circulating
glucose), and all the sequelae which result from diabetes
(18,19).
Glycogen synthesis is also stimulated by the insulin
receptor via IRS-1. In this case, it is the SH2 domain of
PI-3 kinase (PI-3K) that binds the P-Tyr of IRS-1. Now
activated, PI-3K can convert the membrane lipid phosphatidylinositol 4, 5-bisphosphate (PIP2) to phosphatidylinositol 3, 4, 5-triphosphate (PIP3). This indirectly activates
a protein kinase, PKB, via phosphorylation. PKB then
phosphorylates several target proteins, including glycogen
synthase kinase 3 (GSK-3). GSK-3 is responsible for phosphorylating (and thus deactivating) glycogen synthase.
When GSK-3 is phosphorylated, it is deactivated, and
prevented from deactivating glycogen synthase. In this
roundabout manner, insulin increases glycogen synthesis
(20,24).
Once an insulin molecule has docked onto the receptor
and effected its action, it may be released back into the
extracellular environment or it may be degraded by the cell.
Degradation normally involves endocytosis of the insulinreceptor complex followed by the action of insulin degrading
enzyme. Most insulin molecules are degraded by liver cells.
It has been estimated that a typical insulin molecule is
finally degraded about 71 minutes after its initial release into
circulation (28,31).
An English study shows that abdominal obesity
impaired NEFA suppression after oral glucose, and fasting

hyperinsulinemia were present in Sikh MI patients and

their nondiabetic first-degree relatives compared with
Sikh control subjects. PS survivors of premature MI demonstrated impaired insulin-mediated glucose disposal and
NEFA suppression compared with ethnic control subjects.
BWMI patients showed abnormalities of carbohydrate, but
not of NEFA, metabolism compared with white control
subjects. Defects of insulin action manifested as abdominal
obesity, impaired NEFA suppression, and fasting hyperinsulinemia are present in Sikh MI patients and their asymptomatic, nondiabetic, first-degree relatives. We suggest that
these defects may be early metabolic markers that predict
risk of premature MI among PSs (33,34).
The insulin receptor was evaluated at different disease
stages in the sand rat (Psammomys obesus), a model for
nutrition-induced diabetes. Nondiabetic sand rats showed
markedly low receptor number in liver compared with
albino rats. Their receptor had an intact tyrosine kinase
activity but a higher Km for ATP in the phosphorylation
reaction of exogenous substrates. The initial effects of
overeating (i.e., development of hyperinsulinemia without
hyperglycemia) were associated in the sand rat with a
dramatic decrease in vitro and in vivo insulin-induced
receptor tyrosine kinase activity in both liver and muscle.
In muscle, this coincided with a decrease in receptor
number and an increase in basal tyrosine kinase activity.
Similar changes were observed upon development of
hyperinsulinemia with hyperglycemia. Upon recovery
from the diabetic state by diet restriction, the impaired
receptor kinase activation was corrected. Complete
restoration occurred only in animals that fully recovered
from the diabetic state and became normoinsulinemic.
These observations indicate that loss and gain of receptor
tyrosine kinase activity were dependent on insulin levels
(35,36).
Thus, overeating may lead to the development of
hyperinsulinemia through ineffective extraction of excess
insulin by the scarce liver receptors. Hyperinsulinemia, in
turn, causes a reversible reduction in receptor kinase
activity, leading to insulin resistance. This sequence of
events may be relevant to diet-related changes in human
non-insulin-dependent diabetes mellitus.

Archives of the Balkan Medical Union

September 2014, 279

C ONCLUSIONS
1. The insulin receptor function is correlated with
fasting insulinaemia and 1h after OGTT, for all subgroup of study.
2. The main characteristic of sub-groups is grade of
obesity quantified by BMI (body mass index).
3. By comparing BMI with DAM we obtained a direct
correlation, the highest BMI the largest DAM.
4. There is a negative correlation between DAM and
IRF, the largest DAM the lowest affinity of insulin
receptor.

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