Relation of phase angle tertiles with blood adipocytokines levels

advertisement
European Review for Medical and Pharmacological Sciences
2010; 14: 521-526
Relation of phase angle tertiles with blood
adipocytokines levels, insulin resistance and
cardiovascular risk factors in obese women patients
D.A. DE LUIS, R. ALLER*, E. ROMERO, A. DUEÑAS, J.L. PEREZ CASTRILLON
Institute of Endocrinology and Nutrition, Medicine School and Unit of Investigation, Hospital Rio
Hortega; Hospital Clinico*, University of Valladolid, Valladolid (Spain), RETICEF RD 056/0013
Abstract. – Background: Few studies
have evaluated the relation between phase angle
(PA) and metabolic syndrome. As long as we
know, there are not studies of association between phase angle and adipocytokines. The aim
of our study was to evaluate the association of
adipocytokines levels and classical cardiovascular risk factors with tertiles of phase angle in
obese women.
Material and Methods: A cross-sectional
study was designed to establish whether phase
angle from 228 adult female patients with obesity are related with adipocitokynes and cardiovascular risk factors. These patients were studied in a Nutrition Clinic Unit after signed informed consent. All patients with a 2 weeks
weight-stabilization period before recruitment
were enrolled. Weight, blood pressure, basal glucose, C-reactive protein (CRP), insulin, total cholesterol, LDL-cholesterol, HDL-cholesterol,
triglycerides blood and adypocitokines (leptin,
adiponectin, resistin Interleukin-6 and TNF-alpha) levels were measured. The phase angle α
was determined by bioimpedance with the equation [PAº=(Xc/R)x(180º/π)].
Results: Two hundred and twenty-eight females gave informed consent and were enrolled in the study. The mean age was 38.2±14.7
years and the mean BMI 35.27±6.5. Patients
were divided by tertiles of phase angle. Fat
mass was higher in first tertile than third tertile
(43.6±12.6 vs 40.9±15 kg: p<0.05). HOMA
(2.4±1.6 vs 1.46±1.6: p<0.05), insulin (14.4±8.5
vs 11.3±9.4 mUI/L: p<0.05) and glucose
(102.1±20 vs 90±19.5 mg/dl: p<0.05) levels were
higher in first tertile than second and third tertiles. Leptin (167.3±98 vs 104.5±80 ng/ml:
p<0.05) and IL-6 (3.84±5.7 vs 1.8±2.9 pg/ml:
p<0.05) levels were higher in first phase angle
tertile than third tertile phase angle.
Conclusion: Obese women with a low PA tertile have high fat mass with a secondary high
level of glucose, HOMA, IL-6 and leptin. Perhaps,
a low tertile of phase angle could be a new subrogate cardiovascular risk factor to categorize
the obese patients.
Key Words:
Adipocytokines, Cardiovascular risk factors, Obesity,
Insulin resistance, Phase angle α.
Introduction
Whole-body impedance represented by Z vector is a combination of resistance (R) and reactance (Xc). Impedance is a measurable property
of electric ionic conduction of soft tissue of electric ionic conduction of soft tissue; fat and bone
are poor conductors1. In bioelectrical impedance
analysis (BIA), the arc tangent of Xc/R is called
the phase angle2. Phase angle alpha can be easily
obtained from BIA, this angle is one of best single predictor of survival in HIV infected
patients3. Interestingly, the phase angle of the impedance vector was lower because of a decreased
Xc component patients with hemodynamic instability 4, in hemodialysis patients with poorer
prognosis5 and in critically ill patients6.
Obesity and insulin resistance are associated
with cardiovascular risk factors, including altered
levels of inflammatory markers and adipocytokines7. The incidence of obesity and associated
comorbidities is dramatically increasing worldwide. Obesity is characterized by a low grade
systemic inflammation. Epidemiologic evidence
of this rising tide of obesity and associated
pathologies has led, in the last years, to a dramatic increase of researches on the role of adipose
tissue as an active participant in controlling the
body’s physiologic and pathologic processes8.
Adipocytokines are proteins produced mainly by
adipocytes9. These molecules have been shown
to be involved in the pathogenesis of the meta-
Corresponding Author: Daniel A. de Luis, MD; e-mail: [email protected]
521
D.A. de Luis, R. Aller, E. Romero, A. Dueñas, J.L. Perez Castrillon
bolic syndrome and cardiovascular disease. Few
studies have evaluated the relation between phase
angle and metabolic syndrome. Mara et al. 10
showed a significant predictor of phase angle in
the metabolic rate in severely obese patients.
Guida et al. 11 have demonstrated that obese
women with an altered body composition can be
identified and monitoring using bioimpedance.
As long as we know, there are not studies of association between phase angle and adipocytokines.
The aim of our study was to evaluate the association of adipocytokines levels and clasical cardiovascular risk factors with tertiles of phase angle in obese women.
Materials and Methods
Study Design
A cross-sectional study was designed to establish whether phase angle from 228 adult female
patients with obesity are associated with
adipocytokines and cardiovascular risk factors.
These patients were studied in a Nutrition Clinic
Unit after signed informed consent. All patients
with a 2 weeks weight-stabilization period before
recruitment were enrolled. Weight, blood pressure, basal glucose, lipoprotein (a), C-reactive
protein (CRP), insulin, total cholesterol, LDLcholesterol, HDL-cholesterol, triglycerides blood
and adypocitokines (leptin, adiponectin, resistin
and TNF-alpha) levels were measured. Exclusion
criteria included; severy impaired hepatic function (total bilirubin concentration >3.5 mg/dl)
and renal function (serum creatinine concentration >2.5 mg/dl), ongoing infections, major gastrointestinal disease, autoimmune disorders,
steroids treatment, and medication could modulate body composition or weight.
Biochemical Parameters
Serum total cholesterol and triglyceride concentrations were determined by enzymatic colorimetric assay (Technicon Instruments, Ltd.,
New York, NY, USA), while HDL cholesterol
was determined enzymatically in the supernatant
after precipitation of other lipoproteins with dextran sulfate-magnesium. LDL cholesterol was
calculated using Friedewald formula. Plasma
glucose levels were determined by using an automated glucose oxidase method (Glucose analyser
2, Beckman Instruments, Fullerton, CA, USA).
522
Insulin was measured by enzymatic colorimetry
(Insulin, WaKo Pure-Chemical Industries, Osaka,
Japan) and the homeostasis model assessment
(HOMA) for insulin sensitivity was calculated
using these values12.
Adipocytokines
Resistin was measured by ELISA (Biovendor
Laboratory, Inc., Brno, Czech Republic) with a
sensitivity of 0.2 ng/ml with a normal range of 412 ng/ml. Leptin was measured by ELISA (Diagnostic Systems Laboratories, Inc., Texas, USA)
with a sensitivity of 0.05 ng/ml and a normal
range of 10-100 ng/ml. Adiponectin was measured by ELISA (R&D systems, Inc., Minneapolis, MN, USA) with a sensitivity of 0.246 ng/ml
and a normal range of 865-21424 ng/ml. Interleukin 6 and TNF-alpha were measured by
ELISA (R&D systems, Inc., Minneapolis, MN,
USA) with a sensitivity of 0.7 pg/ml and 0.5
pg/ml, respectively. Normal values of IL-6 was
(1.12-12.5 pg/ml) and TNF-alpha (0.5-15.6
pg/ml).
Anthropometric Measurements
Body weight was measured to an accuracy of
0.5 kg and body mass index (BMI) computed as
body weight/(height2). Waist (narrowest diameter
between xiphoid process and iliac crest) and hip
(widest diameter over greater trochanters) circumferences to derive waist-to hip ratio (WHR)
were measured, too. Tetrapolar body electrical
bioimpedance was used to determine body composition13. An electric current of 0.8 mA and 50
kHz was produced by a calibrated signal generator (Biodynamics Model 310e, Seattle, WA,
USA) and applied to the skin using adhesive
electrodes placed on right-side limbs. After introduction of an 800 mA (50 kHz) excitation current, BIA measures the geometrical components
of electrical impedance Zc, i.e., resitance R (the
sum of in phase vectors) and the capacitive component, reactance X (the sum of out-phase vectors) derived from Z2=R2+Xc2. The phase angle α
is determined by the equation [PAº=(X c/R) ×
(180º/π)]. Precautions taken to insure valid BIA
measurements were: no alcohol within 24 hours
of taking the test, no exercise or food for four
hours before taking the test.
Dietary Intake
Patients received prospective serial assessment
of weight, nutritional intake with 3 days written
food records. All enrolled subjects received in-
Phase angle and obesity
struction to record their daily dietary intake for
three days including a week end day. Handling of
the dietary data was by means of a personal computer equipped with personal software, incorporating use of food scales and models to enhance
portion size accuracy. Records were reviewed by
a registered dietitian and analyzed with a computer-based data evaluation system. National
composition food tables were used as reference14.
Drinking and smoking habit were recorded as dicotomic variables.
Statistical Analysis
The results were expressed as means ± standard deviation. The distribution of variables was
analyzed with Kolmogorov-Smirnov test. Quantitative variables with normal distribution were
analyzed with a two-tailed, paired Student’s-t test
and ANOVA test. Non-parametric variables were
analyzed with the Friedman and Wilcoxon tests.
Qualitative variables were analyzed with the chisquare test, with Yates correction as necessary,
and Fisher’s test. Pearson test and Spearman’S
test were used to assess correlation analysis. A pvalue under 0.05 was considered statistically significant.
Results
Two hundred and twenty-eight females gave
informed consent and were enrolled in the study.
The mean age was 38.2±14.7 years and the mean
BMI 35.27±6.5. Baseline characteristics of patients were presented in Table I.
All subjects were weight stable during the 2
weeks period preceding the study (body weight
change, 0.3±0.09 kg). Anthropometric measure-
Table I. Clinical and epidemiological characteristics of
study population.
Characteristics
Age (years)
BMI (kg/m2)
Systolic BP (mmHg)
Diastolic BP (mmHg)
Glucose (mg/dl)
Total cholesterol (mg/dl)
LDL-cholesterol (mg/dl)
HDL-cholesterol (mg/dl)
CRP (mg/dl)
Insulin (UI/L)
HOMA
Values
38.2 ±1 4.7
35.3 ± 6.5
126. 3 ± 17
81.9 ± 12.8
98.1 ± 18
207.5 ± 43.1
123.9 ± 41
57 ± 1
6.6 ± 9.9
15.2 ± 8.6
2.4 ± 1.6
BP: blood pressure. CRP: C-reactive protein. HOMA:
homeostatic model assessment [glucose (mmol/L) x insulin
(µU/ml)/22.5].
ments showed an average waist circunference
(109.2±14 cm), waist-to hip ratio (0.89±0.9), and
average weight (89.5±17.8 kg). Tetrapolar body
electrical bioimpedance showed the next data: fat
free mass (FFM) (45.8±14.4 kg) and fat mass
(43.1±13.5 kg). Serial assessment of nutritional
intake with 3 days written food records showed a
caloric intake of 1672±488 kcal/day, a carbohydrate intake of 169.4±64 g/day, a fat intake of
73.5±13 g/day and a protein intake of 83.9±22.9
g/day.
Anthropometric variables by phase angle tertile groups (tertile 1: phase angle below 6.17º),
(tertile 2: phase angle between 6.18º and 7.07º)
and (tertile 3: phase angle above 7.08º) are
shown in Table II. Average of age was similar in
all tertiles. Fat mass was higher in first tertile
than third tertile and FFM was lower in the first
tertile than third tertile.
Table II. Anthropometric characteristics by tertiles of phase angleº.
Phase angle
Characteristics
Weight (kg)
BMI (kg/m2)
Fat free mass (kg)
Fat mass (kg)
Waist circumference
Waist to hip ratio
(1) (n = 75)
(2) (n = 76)
(3) (n = 77)
89.2 ± 18
35.1 ± 6.4
44.8 ± 6.5
43.6 ± 12.6
106.6 ± 15
0.89 ± 0.08
88.8 ± 15.2
35 ± 5.7
45.7 ± 4.6
42.2 ± 12.5
105.6 ± 13.5
0.89 ± 0.07
88.9 ± 18
35.5 ± 7.1
47.4 ± 9.9*
40.9 ± 15*
105.7 ± 15
0.89 ± 0.08
BMI: body mass index. *(p<0.05), significant difference between first and third ALT tertiles.
523
D.A. de Luis, R. Aller, E. Romero, A. Dueñas, J.L. Perez Castrillon
Table III. Cardiovascular risk factors by tertile of phase angleº.
Phase angle
Characteristics
Systolic BP (mmHg)
Diastolic BP (mmHg)
Glucose (mg/dl)
Total Ch. (mg/dl)
LDL Ch. (mg/dl)
HDL Ch. (mg/dl)
Triglycerides (mg/dl)
Insulin (mUI/L)
CRP (mg/dl)
HOMA-IR
(1) (n = 75)
(2) (n = 76)
(3) (n = 77)
130.1 ± 16
81.4 ± 1
102.1 ± 20
206.1 ± 50
28.6 ± 45
58.1 ± 13
117.9 ± 46
14.4 ± 8.5
6.3 ± 6.4
2.4 ± 1.6
124.5 ± 18
79.7 ± 11
93.8 ± 14
202.4 ± 44
118.6 ± 44
56.8 ± 12
113 ± 58
13.1 ± 7.4
7.1 ± 4.6 3
2.37 ± 11.3
126.9 ± 19.2
81 ± 11
90 ± 19.5*
213.5 ± 35.9
128.3 ± 34
54.7 ± 13
124.8 ± 60
11.3 ± 9.4*
5.6 ± 6.1
1.46 ± 1.6*
*Statistical differences with first tertile (p<0.05) BP: blood pressure; Ch.: cholesterol; HOMA-IR: homeostatic model assessment [glucose (mmol/L) x (nsulin (µU/ml)/22.5].
Cardiovascular risk factors variables by phase
angle tertiles are shown in Table III. HOMA,
insulin and glucose levels were higher in first
tertile than second and third tertiles. Caloric
intake, smoking habit and physical activity
were similar in the three phase angle tertiles
(Table IV).
Table V shows the blood adipocytokines levels. Leptin and IL-6 levels were higher in first
phase angle tertile than third tertile phase angle.
Correlation analysis shows the next associations with phase angle, with FFM (r=0.3;
p<0.05), fat mass (r=–0.13; p<0.05), leptin
(r=–0.2; p<0.05), IL-6 (r=–0.13; p<0.05). Other
parameters that did not reach statistical differences in tertile analysis show significative correlations with phase angle, for example;
adiponectin (r=–0.15; p<0.05), resistin (r=0.1;
p<0.05) and TNF alpha (r=0.1; p<0.05).
Discussion
In our study, we have documented a different
fat mass, glucose, HOMA, leptin and IL-6 levels
in different phase angle tertiles.
Phase angle α (PA), which reflects changes in
electrical conductivity of the body (indicating either alterations in body composition per se or –
more likely – alterations of membrane cell integrity, function, or specific composition, including changes in the intercellular space). Phase angle α can be easily obtained from tetrapolar BIA,
which is a noninvasive and safe technique for
measuring human electrical tissue conductivity15,16. The overall opposition that a body presents to an alternative electric current has two
components. The first is the capacitive reactance
(X), resistive effect produced by the tissue interfaces and cell membranes. The second is the re-
Table IV. Dietary intake and habits by tertile of phase angleº.
Phase angle
Characteristics
Energy (kcal/d)
Carbohydrate (g/d)
Fat (g/d)
Cholesterol (mg/d)
Protein (g/d)
% smoking habit
Hours. aerobic exercise per week
No statistical differences.
524
(1) (n = 75)
(2) (n = 76)
(3) (n = 77)
1670 ± 440
172.7 ± 60
65.2 ± 23.8
355 ± 144
83.8 ± 26
7.7
0.8 ± 2.1
1762 ± 500
70.1 ± 62
76.1± 31
383 ± 119
88.6 ± 24.7
7.5
0.7 ± 1.6
1645 ± 568
165.9 ± 69
73.4 ± 33.1
343 ± 110
88.4 ± 21
7.6
0.8 ± 1.8
Phase angle and obesity
Table V. Circulating adipocytokines by phase angle tertileº.
Phase angle
Characteristics
IL-6 (pg/ml)
TNF-alpha (pg/ml)
Adiponectin (ng/ml)
Resistin (ng/ml)
Leptin (ng/ml)
(1) (n = 75)
(2) (n = 76)
(3) (n = 77)
3.84 ± 5.7
4.3 ± 3.1
32.2 ± 36
3.3 ± 1.5
167.3 ± 98
3.34 ± 1.8
5.3 ± 3.7
25.7 ± 18
3.4 ± 1.1
157.6 ± 82
1.8 ± 2.9*
5.4 ± 2.7
26.7 ± 21.6
3.5 ± 1.7
104.5 ± 80*
IL-6: interleukin 6. *Statistical differences with first tertile (p<0.05).
sistance (R), the restriction to the flow of an electrical current through the body, primarily related
to the amount of water present in the tissues. Part
of the electric current is stored by the cell membranes, which acts as capacitors, creating a phase
shift, quantified geometrically as PA.
Although the biological meaning of PA is not
completely understood17,18, it reflects not only
body cell mass (BCM), but is also one of the best
indicators of cell membrane function, related to
the ratio between extracellular water and intracellular water. Once PA is representative of BCM
and the function of cell membrane, some changes
are expected to occur in its values as a result of
sex. PA is smaller in women than men19. All our
patients are women; effects of sex could no explain our data. Our results showed a relation
among PA and glucose, insulin resistance, leptin
and IL-6 levels in tertile analysis. This relationship include more parameters (adiponectin, resistin, TNF-alpha). Perhaps, a low tertile of
phase angle could be a new subrogate cardiovascular risk factor. However, correlation analysis
shows a linear relationship with other cytokines.
Adipocytokines are proteins produced mainly
by adipocytes, related with fat mass. These molecules have been shown to be involved in the
pathogenesis of the metabolic syndrome and cardiovascular disease. Adiponectin is an adipocytederived collagen like protein indentified through
an extensive search of adipose tissue. Hypoadiponectinemia increased risk of coronary artery
disease togheter with the presence of multiple
risk factors, indicating that adiponectin may be a
key factor of the metabolic syndrome20. Leptin is
a 16 KD protein secreted primarily from
adipocytes. Leptin suppresses food intake and increase energy expenditure by enhancing thermogenesis and metabolic rate. Recent reports suggest that leptin contributes to atherosclerosis and
cardiovascular disease in obese patients21. Resistin is a 12.5 KD, cysteine-rich protein indentified by screening for the genes that are induced
during the differentiation of the adipocytes.
However, the role of resistin in linking human
obesity with type 2 diabetes mellitus is thus
questionable22. Interleukin 6 is increased in most
animal and humans models with obesity and insulin resistance23-24.
In conclusion, obese women with a low PA
tertile have high fat mass with high levels of glucose, HOMA, IL-6 and leptin. Perhaps, a low tertile of phase angle could be a new subrogate cardiovascular risk factor to categorize our obese
patients.
References
1) HEYMSFIELD SB, WANG Z, VISSER M, GALLAGHER D,
PIERSON RN Jr. Tecniques used in the measurement of body composition: an overview with emphasis on bioelectrical impedance analysis. Am J
Clin Nutr 1996; 64(3 Suppl): 478S-484S.
2) FOSTER KR, LUKASKI HC. Whole-body impedance:
what does it measure? Am J Clin Nutr 1996: 64(3
Suppl): 388S-396S.
3) OTT M, FISCHER H, POLAT H, HELM EB, FRENZ M. Bioelectrical impedance analysis as a predictor of
survival in patients with human immunodeficiency
virus infection. J Acquir Immune Defic Syndr Hum
Retrovirol 1995; 9: 20-25.
4) PICCOLI A. Identification of operational clues to dry
weight prescription in hemodialysis using bioimpedance vector analysis. Kidney Int 1995; 53:
1036.
5) CHERTOW GM, LOWRIE EG, LEW NL, LAZARUS JM. Bioelectrical impedance analysis predicts survival in
hemodialysis patients. J Am Soc Nephrol 1996; 7:
1442.
525
D.A. de Luis, R. Aller, E. Romero, A. Dueñas, J.L. Perez Castrillon
6) SCHELTINGA MR, KIMBROU TD, JACOBS DO, WILMORE
DW. Altered cell membrane function in critical illness can be characterized by measuring body reactance. Surg Forum 1990; 41: 43-44.
7) FANTUZZI G. Adipose tissue, adipokines, and inflammation. J Allergy Clin Immunol 2005; 115;
911-919.
8) MEIER U, GRESSNER AM. Endocrine regulation of energy metabolism: review of pathobiochemical and
clinical chemical aspects of leptin, ghrelin,
adiponectin and resistin. Clin Chem 2004; 50:
1511-1525.
15) KUSHNER RF, SCHÖLLER DA. Estimation of total body
water by bioelectrical impedance analysis. Am J
Clin Nutr 1986; 44: 417-424.
16) SLUYS TEMS, VAN DER ENDE ME, SWART GR. Body
composition in patients with acquired immunodeficiency syndrome: a validation study of bioelectric impedance analysis. J Parenter Enter Nutr
1993; 17: 404-406.
17)
DE LUIS DA, ALLER R, IZAOLA O, BACHILLER P, GONZALEZ
SAGRADO M. Influence of hormonal status and oral
intake on phase angle in HIV infected patients.
Nutrition 2004; 20: 731-734.
9) M AT S U D A M, S H I M O M U R A I, S ATA M. Role of
adiponectin in preventing vascular stenosis. The
missing link of adipo-vascular axis. J Biol Chem
2002; 277: 37487-37491.
18) DE LUIS DA, ALLER R, IZAOLA O, TERROBA MC, CABEZAS
G, CUELLAR L. Tissue electric properties in head
and neck cancer patients. Ann Nutr Met 2006: 50:
7-10.
10) MARA M, PASANISI F, SCALFI L. The prediction of basal
metabolic rate in young adult, severely obese patients using single frequency bioimpedance
analysis. Acta Diabetol 2003; 40: s139-s142.
19) KYLE UG, GENTON L, SLOSMAN DO, RICHARD C. Fat
free and fat mass percentiles in 5225 healthy
subjects aged 15 to 98 years. Nutrition 2001; 17:
534-541.
11) GUIDA B, TRIO R, PECORARO P, GERARDI MC, LACCETTI
R, NASTASI A, FALCONI C. Impedance vector distribution by body mass index and conventional bioelectrical impedance analysis in obese women.
Nutr Metab Cardiovasc Dis 2003; 13: 72-79.
20) KUMADA M, KIHARA S, SUMITSUJI S. Association of hypoadiponectinemia with coronary artery disease
in men. Arterioscler Thromb Vasc Biol 2003; 23:
85-89.
12) MATTHEWS DR, HOSKER JP, RUDENSKI AS, NAYLOR BA,
TREACHER DF, TURNER RC. Homesotasis model assessment: insulin resistance and beta cell function from fasting plasma glucose and insulin concentrations in man. Diabetologia 1985; 28: 412419.
13) PICCOLI A, BRUNANI A, SAVIA G, PILLON L, FAVARO E,
BERSELLI ME, CAVAGNINI F. Discriminating between
body fat and fluid changes in the obese adult using bioimpedance vector analysis. Int J Obesity
1998; 22: 97-104.
14) MATAIX J, MAÑAS M. Tablas de composición de alimentos españoles. Ed: University of Granada,
2003.
526
21) SHIMOMURA I, HAMMER RE, IKEMOTO S. Leptin reverses insulin resitance and diabetes mellitus in mice
with congenital lipodystrophy. Nature 1999; 401:
73-76.
22) STEPPAN CM, BAILEY ST, BHAT S. The hormone resistin links obesity to diabetes. Nature 2001; 409:
307-312.
23) MATSUZAWA Y. Adipocytokines: emerging therapeutic targets. Curr Atheroscler Rep 2005; 7: 58-62.
24)
DE LUIS DA, ALLER R, IZAOLA O, GONZALEZ SAGRADO,
CONDE R, PEREZ CASTRILLON JL. Effects of lifestyle
modification on adipocytokine levels in obese patients. Eur Rev Med Pharmacol Sci 2008; 12: 3339.
Download