Is there a relationship between the increase in leptin, CRP, TNF-α, and NO and the degree of obesity in obese Egyptian adolescents? : Medical Research Journal

Secondary Logo

Journal Logo

Advanced Search
ORIGINAL ARTICLES

Is there a relationship between the increase in leptin, CRP, TNF-α, and NO and the degree of obesity in obese Egyptian adolescents?

El-Wakkad, Amanya; Hassan, Nayera Elmorsib; Sibaii, Hibac; El Zayat, Salwac

Author Information

aDepartments of Clinical Pathology

bBiological Anthropology

cMedical Physiology, National Research Center, Cairo, Egypt

Correspondence to Amany El-Wakkad, Medical Physiology Department, National Research Centre, El-Bohouth Street, Dokki, POB: 2311, Cairo, Egypt Tel: +02 26367178; e-mail: [email protected]

Received July 1, 2011

Accepted October 10, 2011

Medical Research Journal 10(2):p 97-101, December 2011. | DOI: 10.1097/01.MJX.0000407614.32015.7a
  • Free

Abstract

Introduction

Obesity is considered as a low-grade chronic inflammatory condition 1 characterized by elevated circulating levels of leptin 2, C-reactive protein (CRP) 3, and TNF-α 4; these proinflammatory cytokines are also well-known inducers of nitric oxide (NO) overproduction 5. Leptin, the product of ob gene, is an anorexic hormone secreted by adipose tissue, which is primarily known for its role as a central regulator of food intake, body weight, and adipose stores. Circulating leptin concentrations are proportional to the total adipose mass and it decreases after weight loss 6. It may provide a possible connection between body fat and immune function 6. Obese individuals typically have relatively elevated serum leptin levels, suggesting that, in most humans, obesity may be associated with leptin insensitivity rather than with leptin deficiency 7. The study of Jang et al.8 revealed that NO is considered as a factor with a role in the modulation of food intake and obesity, and that NO is a potential regulator of leptin-induced lipolysis 9. Furthermore, a high-fat diet increased the expression of inducible NO synthase (NOS) in various peripheral tissues 5, and the inhibition of NOS in adipose tissue leads to increased lipolysis 10. Also, the study of Choi et al.11 has demonstrated elevated serum levels of NO metabolites (nitrate and nitrite) in obese adolescents, and this increase begins in participants with a BMI greater than 25 kg/m2. Ntyintyane et al.12 stated that leptin is a hormone secreted by the adipose tissue, and has been positively correlated with obesity. This hormone controls body weight through the central regulation of food intake and energy expenditure 13. In addition, Bruno et al.14 showed that the adipocyte-derived hormone leptin may provide a possible connection with the proinflammatory CRP, TNF-α, and NO 15,16. Also, the study of Dixit et al.17 revealed that NO might be crucial in mediating the endocrine effects of leptin in the pituitary gland and the mononuclear cells.

Dube et al.18 have demonstrated that hyperleptinemia in obesity is associated with increased CRP; a positive correlation between CRP and leptin has also been shown previously 19,20. Recent studies have demonstrated that serum concentrations of CRP 21 and leptin 22 were positively associated with BMI. Bastard et al.23 reported that CRP is regulated by multiple cytokines including TNF-α. The aim of this study was to investigate the relation between the degree of obesity and the serum level of leptin, CRP, TNF-α, and NO.

Methods

The current study was carried out on obese Egyptian adolescents of both sexes, with an age range from 13 to 18 years, in the National Research Centre, Egypt, to estimate the prevalence of obesity among school children and adolescents. It was a cross-sectional survey. Four local public schools situated in the Giza governorate were enrolled in this study regarding adolescents (two primary schools and two secondary schools). The study included boys and girls during the period of October 2007–April 2009. Permission to perform the study was granted by the Ministry of Education, and the directors of the school were included in the research.

The protocol was approved by the ‘Ethical Committee’ of the ‘National Research Centre’. Of the total sample, 103 adolescents (22 boys and 81 girls) with the complaint of obesity were included in the current research after obtaining written informed consent from their parents. Student assent was also obtained. The adolescents were required to meet the following inclusion criteria: age 13–18 years, and BMI greater than 95 percentile for age and sex on the basis of Center of Disease Control Charts 24. Adolescents were excluded if they had a prior major illness, including type I or II diabetes, took medications, or had a condition known to influence body composition, insulin action, or insulin secretion (e.g. glucocorticoids therapy, hypothyroidism, Cushing's disease).

Each adolescent underwent a complete physical examination, including anthropometric measures. The height and the weight were recorded. The height was measured to the nearest 0.5 cm on a Holtain portable anthropometer, and the weight was determined to the nearest 0.1 kg on a Seca scale Balance with the participant dressed in minimum clothes and no shoes. The BMI was calculated as the weight (in kilograms) divided by the height (in meters) squared. According to their BMI results, the participating adolescents were divided into two groups according to the BMI percentile for age and sex: group I with BMI greater than 95 percentile and less than 97 percentile; group II with BMI greater than 97 percentile 24. Blood samples was drawn from the obese Egyptian adolescents, and the serum was separated and kept frozen at −70°C until analysis.

Biochemical assays

Serum CRP levels were determined with an enzyme-linked immunosorbant assay (ELISA) technique according to the method of Roberts et al.25, using commercial kits (BioCheck Inc., Foster City, California, USA), and the sensitivity of the detection level was 0.1 mg\l. Serum concentrations of cytokines TNF-α were measured using commercially available ELISA kits (Ani BiotechOy Orgenium Laboratories Business Unit Finland; Bender MedSystems GmbH Campus Vienna Biocenter, Vienna, Austria), and the sensitivity of detection for TNF was 2.3 pg/ml according to the method described by Adolf and Apfler 26. Leptin levels were determined with an ELISA procedure using commercial kits (Diagnostics Biochem Canada Inc., Ottawa, Canada), and the sensitivity of the detection level was 0.5 ng/ml according to the method of Maffei et al.27. Serum levels of NO were measured using a colorimetric nonenzymatic assay for NO product no. NB88 (Oxford Biomedical Research, Oxford, USA, Superior Science Reliable Results); the kit can be used to accurately measure as little as 1 pmol/ml following the method described by manufacturer.

Statistical analysis

All values are expressed as mean±SD and the differences among the two groups were calculated by Student t-test. Correlations were performed between different parameters using Pearson correlation, and P-value <0.05 was considered significant. All analyses were carried out using SPSS version 9.0 (IBM, Chicago, IL, USA) statistical software.

Results

The data for the two groups are shown in Table 1.

T1-8
Table 1:
Showing the comparison between group I and II for age, sex and BMI percentile

As shown in Table 2, the BMI percentile for age and sex (Center of Disease Control BMI for age growth charts) 24 was used to classify obese adolescents into two groups. The level of proinflammatory cytokine TNF-α was significantly increased (P<0.0005) in the very obese participants in group II (BMI>97 percentile) as compared with group I (BMI>95 percentile and <97 percentile). Also, CRP has shown a very high significant increase (P<0.0005) in group II as compared with group I, whereas leptin has shown a significant increase at P<0.0005 in group II in comparison with group I. NO has shown a significant increase at P<0.005 in the very obese participants in group II as compared with group I.

T2-8
Table 2:
Comparison of different anthropmetrical and biochemical parameters between obese Egyptian adolescents in group I and group II

Correlations between BMI and NO, leptin, CRP, and TNF-α in group II are shown in Table 3. A positive correlation between BMI and NO at r=0.824, P<0.0005 was recorded, as shown in Table 3; also, a highly positive correlation between BMI and leptin at r=0.944, P<0.0005 and a positive correlation of BMI with CRP at r=0.981, P<0.0005 were observed. Similarly, a positive correlation between BMI and TNF-α at r=0.953, P<0.0005 was detected. In addition, there was a positive correlation between NO and leptin, CRP, and TNF-α at r=0.816, P<0.0005; r=0.816, P<0.0005; r=0.822, P<0.0005; and r=0.793, P<0.0005, respectively, and leptin with CRP and TNF-α at r=0.970, P<0.0005 and r=0.915, P<0.0005, respectively. Also, a positive correlation between CRP and TNF-α at r=0.944, P<0.0005 has been observed.

T3-8
Table 3:
Correlation Between different parameters in group II

Discussion

The data in the present study revealed that CRP, NO, TNF-α, and leptin were positively correlated with BMI, which was in agreement with Wu et al.28, Choi et al.11, Weichhaus et al.29, and Buettner et al.22, respectively; in addition, participants with BMI greater than 97 percentile had higher serum concentrations of TNF-α, leptin, CRP, and NO than participants with BMI lesser than 97 percentile, which led us to determine whether this increase is associated with the degree of obesity. No similar study has been reported in this concern. However, it has been shown by the study of Chen et al.20 that leptin and high-sensitivity CRP, and TNF-α, are tightly linked, and this was also true in the current study in group II. Leptin is secreted from adipocytes including production of other adipocytokines such as TNF-α that promote high-sensitivity CRP synthesis by the liver 20. In addition, there was an increase in the serum concentration of NO in obese Egyptian adolescents, and there was a significant correlation between BMI and NO, and this was proved by Choi et al.11. These investigators showed that this increase begins in participants with a BMI greater than 95 percentile 11. Therefore, it seems that increased serum concentrations of NO may be a result of its production in adipose tissue. There was also an increase in TNF-α and NO because TNF-α is a mediator of inflammation and it enhances the production of reactive oxygen species including inducible NO 30. Our results showed increased serum concentrations of NO in obese adolescents and a significant positive correlation between serum NO concentration and BMI of obese Egyptian adolescents .These findings were in agreement with the study of Magdalena et al.31 and Choi et al.11. In addition, there was a significant positive correlation between NO and leptin, and this was in agreement with Konukoglu et al.32. This increase was explained by Mehebik et al.16, who demonstrated that NO produced in response to leptin by adipocytes could also be a mediator of some leptin effects on adipose tissue metabolism, adipogenesis, and the physiological concentrations of leptin stimulate NOS activity in adipocytes. NOS activity is a novel target for leptin in adipocytes and leptin-induced NOS activity is at least in part the result of NOS phosphorylations through both protein kinase A and p42/p44 MAPK activation. This led them to hypothesize that NO is a potentially important factor for leptin signaling in adipocytes. However, it has been shown that the increase in CRP concentration has been reported to be significantly related to the degree of obesity 33.

In the present study, CRP was increased significantly in obese adolescents, and this was consistent with the results of Cook et al.34. Das 35 explained this increase in CRP by activating a complement that binds to Fc receptors and leads to the generation of proinflammatory cytokines. These findings together with the correlation between serum concentration of CRP, leptin, NO, and TNF-α with the BMI in the group of obese participants whose BMI was greater than 30 kg/m2 support the hypothesis that higher concentrations of these parameters in group II may be the result of increased synthesis in the adipose tissue and fat cells, which was in agreement with Magdalena et al.31, and that the opposite effects of these substances on the development of adipose tissue may constitute a mechanism regulating further body mass gain 32. In general, the present study showed that there was an increase in the four parameters measured in the study, and also, there was a correlation between CRP, leptin, TNF-α, and NO, and BMI, which led us to conclude that the degree of obesity is responsible for this increase.

F1-8
Figure

Acknowledgements

Conflicts of interest

There are no conflicts of interest.

References

1. Giordano P, Del Vecchio GC, Cecinati V, Delvecchio M, Altomare M, De Palma F, et al. Metabolic, inflammatory, endothelial and haemostatic markers in a group of Italian obese children and adolescents. Eur J Pediatr. 2011;170:845–850
2. Singh M, Bedi US, Singh PP, Arora R, Khosla S. Leptin and the clinical cardiovascular risk. Int J Cardiol. 2010;140:266–271
3. Kassi E, Dalamaga M, Hroussalas G, Kazanis K, Merantzi G, Zachari A, et al. Adipocyte factors, high-sensitive C-reactive protein levels and lipoxidative stress products in overweight postmenopausal women with normal and impaired OGTT. Maturitas. 2010;67:72–77
4. Stanley TL, Zanni MW, Johnsen S, Rasheed S, Makimura H, Lee H, et al. Obesity is associated with activation of the TNF-{alpha}antagonism with etanercept decrease glucose and Increases the proportion of high molecular weight adiponectin in obese subjects with features of the metabolic syndrome. J Endocrinol Metab. 2011;96:E146–E183
5. Perreault M, Marette A. Targeted disruption of inducible nitric oxide synthase protects against obesity-linked insulin resistance in muscle. Nat Med. 2001;7:1138–1143
6. Meyers JA, McTiernan A, Ulrich CM. Leptin and immune function: integrating the evidence. Nutr Res. 2005;25:791–803
7. Friedman JM. The function of leptin in nutrition, weight and physiology. Nutr Rev. 2002;60:S1–S14
8. Jang EH, Park CS, Lee SK, Pie JE, Kang JH. Excessive nitric oxide attenuates leptin-mediated signal transducer and activator of transcription 3 activation. Life Sci. 2007;80:609–617
9. Fruhbeck G, Gomez Ambrosi J. Modulation of the leptin-induced white adipose tissue lipolysis by nitric oxide. Cell Signal. 2001;13:827–833
10. Ryden M, Elizalde M, van Harmelen V, Ohlund A, Hoffstedt J, Bringman S, et al. Increased expression of eNOS protein in omental versus subcutaneous adipose tissue in obese human subjects. Int J Obes Relat Metab Disord. 2001;25:811–815
11. Choi JW, Pai SH, Kim SK, Ito M, Park CS, Cha YN. Increases in nitric oxide concentrations correlate strongly with body fat in obese humans. Clin Chem. 2001;47:1106–1109
12. Ntyintyane L, Panz V, Raal FJ, Gill G. Leptin, adiponectin and high-sensitivity C-reactive protein in relation to the metabolic syndrome in urban South African blacks with and without coronary artery disease. Metab Syndr Relat Disord. 2009;7:243–248
13. Ahima RS, Flier JS. Leptin. Annu Rev Physiol. 2000;62:413–437
14. Bruno A, Conus S, Schmid I, Simon HU. Apoptotic pathways are inhibited by leptin receptor activation in neutrophils. J Immunol. 2005;174:8090–8096
15. Mastronardi CA, Yu WH, McCann SM. Resting and circadian release of nitric oxide is controlled by leptin in male rats. Proc Natl Acad Sci USA. 2002;99:5721–5726
16. Mehebik N, Jaubert AM, Sabourault D, Giudicelli Y, Ribiere C. Leptin-induced nitric oxide production in white adipocytes is mediated through PKA and MAP kinase activation. Am J Physiol Cell Physiol. 2005;289:C379–C387
17. Dixit VD, Mielenz M, Taub DD, Parvizi N. Leptin induces growth hormone secretion from peripheral blood mononuclear cells via a protein kinase C- and nitric oxide-dependent mechanism. Endocrinology. 2003;144:5595–5603
18. Dube MG, Torto R, Kalra SP. Increased leptin expression selectively in the hypothalamus suppresses inflammatory markers CRP and IL-6 in leptin-deficient diabetic obese mice. Peptides. 2008;29:593–598
19. Shamsuzzaman AS, Winnicki M, Wolk R, Svatikova A, Phillips BG, Davison DE, et al. Independent association between plasma leptin and C-reactive protein in healthy humans. Circulation. 2004;109:2181–2185
20. Chen K, Li F, Li J, Cai H, Strom S, Bisello A, et al. Induction of leptin resistance through direct interaction of C-reactive protein with leptin. Nat Med. 2006;12:425–432
21. Brooks GC, Blaha MJ, Blumenthal RS. Relation of C-reactive protein to abdominal adiposity. Am J Cardiol. 2010;106:56–61
22. Buettner R, Bollheimer LC, Zietz B, Drobnik W, Lackner K, Schmitz G, et al. Definition and characterization of relative hypo- and hyperleptinemia in a large Caucasian population. J Endocrinol. 2002;175:745–756
23. Bastard JP, Jardel C, Bruckert E, Blondy P, Capeau J, Laville M, et al. Elevated levels of interleukin 6 are reduced in serum and subcutaneous adipose tissue of obese women after weight loss. J Clin Endocrinol Metab. 2000;85:3338–3342
24. Ogden CL, Carroll MD, Curtin LR, Lamb MM, Flegal KM. Prevalence of high body mass index in US children and adolescents, 2007–2008. JAMA. 2010;303:242–249
25. Roberts WL, Sedrick R, Moulton L, Spencer A, Rifai N. Evaluation of four automated high-sensitivity C-reactive protein methods: implications for clinical and epidemiological applications. Clin Chem. 2000;46:461–468
26. Adolf GR, Apfler I. A monoclonal antibody-based enzyme immunoassay for quantitation of human tumor necrosis factor binding protein I, a soluble fragment of the 60 kDa TNF receptor, in biological fluids. J Immunol Methods. 1991;143:127–136
27. Maffei M, Halaas J, Ravussin E, Pratley RE, Lee GH, Zhang Y, et al. Leptin levels in human and rodent: measurement of plasma leptin and ob RNA in obese and weight-reduced subjects. Nat Med. 1995;1:1155–1161
28. Wu DM, Chu NF, Shen MH, Chang JB. Plasma C-reactive protein levels and their relationship to anthropometric and lipid characteristics among children. J Clin Epidemiol. 2003;56:94–100
29. Weichhaus M, Broom I, Bermano G. The molecular contribution of TNF-α in the link between obesity and breast cancer. Oncol Rep. 2011;25:477–483
30. Das UN. GLUT-4, tumour necrosis factor, essential fatty acids and daf-genes and their role in glucose homeostasis, insulin resistance, non-insulin dependent diabetes mellitus and longevity. J Assoc Physicians India. 1999;47:431–435
31. Magdalena OG, Brbara ZMJ, Aleksander Z. Serum concentrations of nitric oxid, tumor necrosis factor (TNF-alpha) and TNF soluble receptors in women with overwieght and obesity. Metabolism. 2004;53:1268–1273
32. Konukoglu D, Serin O, Turhan MS. Plasma leptin and its relationship with lipid peroxidation and nitric oxide in obese female patients with or without hypertension. Arch Med Res. 2006;37:602–606
33. Weiss R, Dziura J, Burgert TS, Tamborlane WV, Taksali SE, Yeckel CW, et al. Obesity and the metabolic syndrome in children and adolescents. N Engl J Med. 2004;350:2362–2374
34. Cook DG, Mendall MA, Whincup PH, Carey IM, Ballam L, Morris JE, et al. C-reactive protein concentration in children: relationship to adiposity and other cardiovascular risk factors. Atherosclerosis. 2000;149:139–150
35. Das UN. Is obesity an inflammatory condition? Nutrition. 2001;17:953–966
© 2011 Medical Research Journal