Accepted Manuscript
Adipokine and cytokine levels in patients with adrenocortical cancer, subclinical Cushing’s syndrome and healthy controls
Anna Babinska, Mariusz Kaszubowski, Piotr Kmieć, Krzysztof Sworczak
| PII: | S0039-128X(18)30163-6 |
| DOI: | https://doi.org/10.1016/j.steroids.2018.08.011 |
| Reference: | STE 8306 |
| To appear in: | Steroids |
| Received Date: | 6 June 2018 |
| Revised Date: | 29 August 2018 |
| Accepted Date: | 30 August 2018 |
ISSN 0039-128X
ELSEVIER
STEROIDS
Please cite this article as: Babinska, A., Kaszubowski, M., Kmieć, P., Sworczak, K., Adipokine and cytokine levels in patients with adrenocortical cancer, subclinical Cushing’s syndrome and healthy controls, Steroids (2018), doi: https://doi.org/10.1016/j.steroids.2018.08.011
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Adipokine and cytokine levels in patients with adrenocortical cancer, subclinical Cushing’s syndrome and healthy controls.
Anna Babinska1, Mariusz Kaszubowski2, Piotr Kmieć 1, Krzysztof Sworczak1
1. Department of Endocrinology and Internal Medicine, Medical University of Gdansk, Poland
2. Institute of Statistics, Department of Economic Sciences, Faculty of Management and Economics, Gdansk University of Technology, Poland
Word count: 3262
Number of tables: 4
Key words: adipokines, cytokines, Subclinical Cushing Syndrome (SCS), adrenocortical carcinoma (ACC)
Corresponding author and person to whom reprint requests should be addressed: Anna Babinska Department of Endocrinology and Internal Medicine, Medical University of Gdansk ul. Dębinki 7, 80-288 Gdansk, Poland phone: +48 58 349 28 40
fax +48 58 349 28 41 e-mail: a.mail@wp.pl Mariusz Kaszubowski Institute of Statistics, Department of Economic Sciences, Faculty of Management and Economics, Gdansk University of Technology ul. Traugutta 79, 80-233 Gdańsk, Poland
phone: +48 58 347 26 23, mkaszubo@zie.pg.gda.pl
Piotr Kmiec
Department of Endocrinology and Internal Medicine, Medical University of Gdansk ul. Dębinki 7, 80-288 Gdansk, Poland
phone: +48 58 349 28 40, fax +48 58 349 28 41, e-mail: piotrkmiec@gumed.edu.pl Krzysztof Sworczak
Department of Endocrinology and Internal Medicine, Medical University of Gdansk ul. Dębinki 7, 80-288 Gdańsk, Poland
phone: +48 58 349 28 40, fax +48 58 349 28 41, e-mail: ksworczak@gumed.edu.pl
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ABSTRACT
Introduction: In recent years researchers have focused at hormonal activity in Cushing’s syndrome (CS) in connection with metabolic disorders and the role of adipokines and cytokines secreted by the adipose tissue. The aim of the study was to investigate levels of adipokines and cytokines in patients with: subclinical CS (SCS) - in relation to hormonal parameters of hypercortisolemia, and, adrenocortical cancer (ACC).
Materials and methods: The study included 20 SCS as well as 7 ACC patients, and 18 healthy participants. Hormonal activity and serum concentrations of adiponectin, leptin, resistin, tumor necrosis factor alpha (TNFa), interleukin 6 (IL6), and monocyte chemoattractant protein 1 (MCP1), were analyzed.
Results: In SCS patients compared to healthy volunteers a trend toward higher concentrations of all pro-inflammatory cytokines was noted, however, statistically significant differences were only found for TNFa and IL6 (p=0.047 and p=0.028, respectively). Adiponectin concentrations were significantly lower in the SCS group (p=0.006). Serum adipokine and cytokine levels were independent of the presence of diabetes mellitus (DM) and hypertension (HT) in the SCS group. A significant correlation was found between subclinical glucocorticoid secretion and IL6 concentration (Pearson’s r=0.517, p=0.02). Acquired results were independent of BMI. In ACC patients compared to controls higher IL6, TNFa and MCP1 levels were recorded.
Conclusion: It is possible that higher adipokine and pro-inflammatory cytokine concentrations as well as lower anti-inflammatory adiponectin concentrations comprise an additional risk factor of metabolic and cardiovascular diseases in SCS patients. It seems that at least among patients with SCS adipokine and cytokine secretion is independent of hormonal activity (except for IL6).
INTRODUCTION
Adrenal incidentalomas, which were previously considered as hormonally inactive, in many cases exhibit subclinical hormonal activity of the adrenal cortex or medulla [1]. Not only in overt but also subclinical Cushing’s syndrome (CS and SCS, respectively) patients the risk of diabetes mellitus (DM), cardiovascular events and hypertension (HT) is higher than in persons without or with a hormonally inactive adrenal tumor [2]. What has also been reported is that the cardiovascular and metabolic disease risk is elevated among patients with non-functioning adrenal incidentalomas (NFAI) [3-8]. Multiple mechanisms that affect morbidity and mortality in CS, SCS, and, possibly, NFAI include pathology connected with the adipose tissue [2].
Central obesity is one of the hallmarks of CS, and, it has been demonstrated that excess visceral adipose tissue can be linked to cardiovascular and metabolic diseases [2, 8-11]. Central accumulation of adipose tissue causes a negative profile of secretion of adipocytokines, i.e. a pro- inflammatory one. While in CS these pathologies are manifest, in SCS and NFAI data in to date literature are often contradictory [4-6].
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Among factors secreted by the adipose tissue, linked to cardiovascular and metabolic diseases, there are pro-inflammatory cytokines, including interleukin 6 (IL6), tissue necrosis factor alpha (TNFa), resistin, and monocyte chemoattractant protein 1 (MCP-1), as well as adipokines such as leptin and adiponectin (and many other). The former are involved in many pathological processes including inflammation, endothelial damage, atherosclerosis, insulin resistance, HT, and bone remodeling [2, 12]. High IL6 concentrations constitute a risk factor of type 2 DM and myocardial infarction [13-15]. TNFa is mainly expressed in monocytes and macrophages (it is crucial in inflammation and autoimmunity), but also affects lipid metabolism, adipocyte function and insulin signaling. Elevated TNFa levels predispose to cardiovascular complications (i.e. myocardial infarct) [2]. Furthermore, TNFa and IL6 lower adiponectin secretion [14, 16, 17]. Resistin (related to obesity, insulin resistance and inflammation) has been associated with an increased risk of myocardial infarction and ischemic stroke [18-20]. MCP1 is a chemoattractant protein involved in adipose tissue inflammation and favors the development of atherosclerosis and chronic circulatory failure [20, 21, 23].
Adiponectin, an important adipokine, sensitizes cells to insulin and has anti-inflammatory and anti-atherogenic properties [23]. Low adiponectin concentrations have been associated with the risk of cardiovascular disease by many researchers [24 - 27]. A negative correlation between circulating adiponectin levels and obesity (in particular central) as well as type 2 DM is well-established [28].
Leptin, the second well-known adipokine, is a satiety regulator and acts oppositely to adiponectin [29]. A relationship between high leptin concentrations and coronary artery calcification has been demonstrated in 860 healthy people without diabetes of similar age and gender without diabetes or established cardiovascular risk factors [30]. Leptin induced C-reactive protein secretion, which is a well-known marker of cardiovascular disease risk [30].
Apart from their role in cardiovascular and metabolic diseases, adipocytokines have also been associated with the incidence of human neoplasms [31]. Leptin has been shown to stimulate the growth of neoplastic cells in the esophageal, stomach, pancreatic, prostate, ovarian, and lung cancers [32]. In our previous study, higher expression of leptin receptors in ACC tumors was demonstrated [33, 34]. Mechanisms underlying the oncogenic effect of leptin have not been clarified thus far. Adiponectin, on the other hand, is recognized as a protein with antineoplastic activity. Its low concentrations were demonstrated in many human malignancies, mainly in the cancers of the breast, reproductive organs, colon and rectum [7]. In the case of low adiponectin concentrations elevated serum insulin and insulin growth factor 1 (IGF1) concentrations lead to increased proliferation, which also applies to neoplastic cells [31, 35]. Adiponectin is also an angiogenesis inhibitor in endothelial cells [36, 37].
The role of adipocytokines in adrenocortical cancer (ACC), a rare neoplasm, has not been studied sufficiently. Interestingly, in approximately fifty percent of cases, ACC leads to hypercortisolemia (both overt and subclinical). The influence of glucocorticoid excess on the biology
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of this malignancy and the possible effects on the secretory function of adipose tissue have not been elucidated.
In the current retrospective study concentrations of adipokines and pro-inflammatory cytokines secreted by the adipose tissue were investigated among SCS patients and ACC patients. In SCS patients, an association between certain adipocytokines and glucocorticoids was examined, and whether secretion of adipokines and cytokines constitutes an additional etiologic factor of metabolic diseases.
SUBJECTS AND METHODS
Subjects
The study was approved by the local ethical committee. Informed consent was obtained from all participants.
In the study the following participants were enrolled: 18 healthy control subjects, 7 adrenocortical cancer patients, and, 20 patients with an adrenal incidentaloma, who exhibited subclinical hormonal activity, i.e. SCS patients. Controls were comparable for sex, age and BMI with both SCS and ACC patient groups. Subjects with an acute inflammatory condition were excluded from the study. None of the study participants had history of an acute cardiovascular event like stroke, myocardial infarction or embolism. No control subject was diagnosed with HT, DM, cardiovascular disease, chronic inflammatory diseases, chronic hepatitis or malignancy. Among SCS patients, hypertension (HT) was present in 5 (25%) and diabetes mellitus (DM) in 6 (30%) (Table 1). Patients with DM were newly diagnosed and did not receive any antidiabetic treatment. No patient was treated with lipid-lowering or antidiabetic medications for at least 12 weeks prior to the study.
In the control group, an adrenal tumor was excluded on the basis of imaging examinations (CT) performed in the preceding 6 months for reasons other than suspicion of an adrenal lesions (i.e. on participants’ own initiative or due to uncharacteristic abdominal pain). Adrenal incidentaloma diagnosis was based on the detection of a unilateral adrenal mass of 1cm or larger on abdominal imaging performed due to a reason different than investigation for an adrenal disease.
All participants were admitted to our Department and examined on the day of blood collection; anthropometric parameters were obtained: height using a wall-mounted ruler, and, weight using a digital scale. Then, body mass index (BMI) was calculated according to the formula: weight (in kg) divided by the square of the height (in m2).
All medications that interfere with laboratory test results were excluded. Venous blood samples were obtained by venipuncture. Morning samples were drawn between 7.30 and 8.30 am after 12-hour overnight fast. Morning and midnight serum cortisol, and morning plasma corticotropin (ACTH) levels were determined under basal conditions to assess the circadian rhythm of hormone secretion. All patients underwent the overnight 1-mg dexamethasone (DXM) test. Cortisol excretion with urine was assessed by 24-hour urine collection.
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SCS was diagnosed in accordance with the definition, i.e. in patients with an incidental adrenal mass discovered in an abdominal CT or MRI scan, who did not have typical physical features of CS but exhibited endogenous hypercortisolemia in hormonal tests. Cortisol value of at least 140 nmol/l in the overnight suppression test with 1 mg of DXM was a sufficient criterion to diagnose SCS [16, 38, 39]. Patients with cortisol values between 50 and 140 nmol/l in this test had to meet at least one of the following criteria to be diagnosed with SCS: lack of circadian rhythm of cortisol secretion (evening to morning serum cortisol ratio exceeding 50%), decreased morning ACTH concentration (to less than 10 pg/ml), and/or increased free urinary cortisol excretion (UFC). Hormonal characteristics of SCS and ACC patients were shown in Table 2. Hormonal activity was not assessed in the control group.
According to the guidelines for the management of patients with adrenal tumors [16], not all SCS patients require surgical treatment. In case of patients with subclinical hypercortisolemia, annual hormonal assessment and monitoring of the occurrence of diseases potentially associated with excess cortisol (such as DM and HT) is suggested [16]. Based on such assessment, potential benefits resulting from surgery should be taken into account. All SCS patients included in this study underwent resection of adrenal incidentalomas due to: concomitant HT (5 patients) and DM (6 patients), tumor progression in imaging tests (>1 cm per year) in 3 cases, and, patients’ fear of cancer in the remaining 6 cases. Histological examination revealed an adrenal cortex adenoma in 12 cases, and nodular hyperplasia in 8.
Incidentally discovered adrenocortical cancer was suspected in patients with lesions detected by CT or MRI scans that were larger than 4 cm, that had high contrast medium enhancement, and incomplete (<50%) delayed washout in CT scans. All patients with suspected ACC underwent surgery and the diagnosis was confirmed in histopathological examination.
Laboratory analyses
All hormone concentrations were determined in the same laboratory using freely available kits. ACTH concentration was determined with a solid-phase, two-site sequential chemiluminescent immunometric assay Immulite 1000 RACTH manufactured by Siemens (repeatability coefficient of variation, CV: 10%). Serum and urinary cortisol concentrations were determined with a Chemiluminescent Microparticle Immunoassay Cortisol Reagent Kit on Abbott’s Architect analyzer (accuracy CV: 5%, repeatability CV: 3.29%).
Morning serum dehydroepiandrosterone sulphate (DHEAS) and androstendione concentrations were determined by radioimmunological assays (RIA) using commercial kits by Orion Diagnostica and DRG MedTek Elisa, respectively (repeatability CVs: 9.8%, and 9.3%, respectively).
Both in patient (SCS and ACC) and control groups, plasma fasting glucose and insulin were measured. Insulin resistance index HOMA (homeostasis model assessment) was determined. The HOMA index was calculated according to the following formula: fasting glucose (mmol/l) x fasting insulin level (mU/l) / 22.5. HOMA index greater than 2.5 was considered abnormal. In six patients
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diagnosed with DM, glycated hemoglobin (HbA1c) was determined. Total cholesterol levels were determined in study participants.
MTPL-Reader “E-lizaMAT” 3000 manufactured by DRG Diagnostics was used for determining concentrations of adipokines and cytokines. Serum levels of adiponectin were determined using Human HMW Quantikine Elisa Kit (Biokom, R&D Systems USA, intra-assay CV: 2.6-3,7%), leptin using a commercial sandwich enzyme immunoassay EIA kit (DRG MedTek, DRG Instruments GmbH Germany, intra-assay CV 5.95-6.91% ), resistin using a Quantikine Elisa Kit (Biokom, R&D Systems USA, intra-assay CV 3.8-5.3%); TNFa, MCP1, and, IL6 using Quantikine HS Elisa (Biokom, R&D Systems USA, respective ranges of intra-assay CVs: 3.1-8.7%, 4.7-7.8%, 6.9-7.8%). Obtained adipokine and cytokine concentrations were the result of consensus of two independent investigators performing the assay.
Statistical analyses
All raw data for each group were presented with their number and basic descriptive statistics as mean ± standard deviation. Normality of data sets was verified using the W Shapiro-Wilk test. Since not all groups met this assumption, differences between mean values (or medians) in small groups were examined by Welch’s t test (symbol t) or Mann-Whitney U test (symbol U) at the same time. If both tests had the same results the p-value for parametric t-test was presented. Differences in fractions were checked using Fisher’s exact test. The level of significance was set at a=0.05. All calculated p-values were for two-tailed tests. Statistical analysis was performed with Statistica 13.1 software (StatSoft, Tulsa, OK, US).
RESULTS
This study included 20 patients with SCS (SCS group), 7 patients with ACC (ACC group), and 18 healthy volunteers (control group). Demographic and clinical features of study participants were presented in Table 1. There were no significant differences in age, sex, and BMI between either patient group and controls.
DM and HT was present only in SCS patients (30% and 25%, respectively). Systolic and diastolic blood pressure values were significantly higher in SCS patients than controls (p=0.001; p<0.001 respectively). Mean HbA1c concentration of six patients with DM was 6.73±0.25% (range: 6.4-7.1%). There were no differences in fasting insulin and HOMA indices between groups (Table 1).
Adipokine and cytokine concentrations were compared between SCS patients and controls. There was a trend toward higher median concentrations of all studied adipokines and pro- inflammatory cytokines, i.e. leptin, resistin, TNFa, IL6, and, MCP1, in the SCS group, although statistically significant differences were only found for IL6 and TNFa levels (U p=0.028 and U p=0.013, respectively). The difference between median leptin concentrations of SCS patients and
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controls bordered statistical significance (U p=0.063). Conversely, median adiponectin concentrations were higher in healthy volunteers than in patients with SCS (Table 3).
Concerning hormonal parameters of glucocorticoid excess, the 1 mg DMX test has the greatest value in the diagnosis of subclinical hypercortisolemia. In SCS patients a moderate positive correlation between cortisol concentrations in the overnight DXM test and IL6 was found: Pearson’s correlation coefficient r equaled 0.517, p=0.02. Morning serum and 24-hour UFC concentrations negatively correlated with leptin concentrations in these patients: Pearson’s correlation coefficients equaled - 0.471 (p=0.036) and -0.479 (p=0.033), respectively. There were no other statistically significant relationship between hormonal parameters and adipokine as well as cytokine levels among SCS patients.
The authors tested potential correlations between cytokines and adipokines, and, the presence of HT and DM in SCS patients; however, no statistically significant correlations were found.
Concentrations of adipokines and cytokines were also compared between ACC patients and controls. Statistically higher IL6, TNFa, and, MCP1 levels were found in the former group (Table 4). Small size of the ACC group did not allow for a statistical analysis of adipokine and cytokine levels and mortality or recurrence-free survival. A correlation between ACC tumor size, which is an important prognostic factor in these patients, and concentrations of adipokines and cytokines was verified -obtained results did not reach statistical significance (probably due to small sample size).
DISCUSSION
Authors of many reports found that glucocorticoid hypersecretion, CS and SCS, leads to increased risk of metabolic diseases and cardiovascular events [1, 4, 7-10, 13]. In the current study, the authors examined 20 SCS and 18 controls comparable for BMI, which eliminates obesity as a potential factor influencing adipokine and cytokine secretion. HT and DM was present solely in 25% and 30% of SCS patients, respectively. No associations between adipocytokine levels and DM as well as HT were found among patients with SCS.
In our study, a trend toward higher mean concentrations of all pro-inflammatory cytokines and adipokines in SCS patients compared to healthy volunteers was apparent, although statistically significant differences were only recorded for TNFa and IL6. In reports by other researchers, overt glucocorticoid excess was associated with increased concentration of pro-inflammatory cytokines, including IL6 [2, 14, 15]. In female CS patients IL6 was significantly higher compared to the control group of women of comparable age and BMI. High levels of IL6 persisted in this group of patients up to 11 years after CS had been cured [2]. Abnormally high levels of IL6 in CS play a role in the pathogenesis of vascular and metabolic complications associated with chronic hypercortisolemia. So far, similar studies have not been conducted in SCS patients. In three observational studies TNFa level was normal among CS patients and did not correlate with cortisol levels, whereas Barahona et al [40] found that TNFa soluble receptor was significantly higher in CS patients compared to controls.
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Concerning resistin, Krsek at al [41] showed that in CS patients concentrations of this cytokine were elevated and positively associated with BMI, but not with UFC or insulin. Of note, Emertici et al reported significantly higher resistin levels in adrenal incidentaloma patients versus controls [4]. These reports stand in contrast to our results. So far, MCP1 concentrations in adrenal tumors have not been widely studied. In vitro it was shown glucocorticoids inhibit MCP1 secretion [21]. In the current study only a trend toward higher MCP1 levels was observed in the SCS group versus controls, and, no associations between MCP1 and cortisol secretion parameters were found.
In contrast to elevated pro-inflammatory cytokines’ concentrations in SCS, adiponectin was higher in healthy volunteers (this finding is independent of participants’ BMI). Similarly, Dogruk Unal and colleagues demonstrated that low adiponectin levels are a valuable indicator of SCS in adrenal incidentaloma patients [24]. Although there are studies, in which a relationship between adiponectin and glucocorticoids was studied, their results are inconclusive due to complex regulation of adiponectin expression [28, 42]. In this study, no correlation was found between adiponectin levels and: cortisol secretion as well as incidence of HT and DM among SCS patients.
In previous studies, circulating leptin levels were higher in CS patients than in healthy controls with normal body weight [43]. Weise found a positive correlation between BMI and leptin levels in CS patients [44]. Interestingly, it is speculated that increase in leptin is a compensatory mechanism that antagonizes excess glucocorticoids [2]. However, so far, the relative amount of visceral versus subcutaneous fat has not been investigated in CS (or SCS) patients, which is crucial in investigating leptin levels, since the latter tissue type secretes two- to three-fold more adipokine than the former [45]. Leptin concentrations were higher in SCS patients compared to healthy controls examined by us. Despite this fact, negative correlations between leptin and UFC as well as morning serum cortisol were found, which calls for further studies of the crosstalk between glucocorticoids and leptin secretion.
Wagenmakers and colleagues showed that adverse adipokine profile (i.e. high levels of leptin and resistin accompanied by low adiponectin levels) persisted for a long time after curing CS patients, which was attributed to persisting central distribution of adipose tissue [11]. Roerink and co-workers demonstrated a relationship between the size of adipocytes and the severity of hypercortisolemia in CS patients [46]. Authors of the current study propose that in SCS even mild but chronic excessive levels of glucocorticoids lead to central adipose tissue accumulation. This, in turn, is connected with an adverse adipokine and cytokine profile that comprises an additional factor for metabolic complications in SCS patients.
There are only few studies that evaluate the role of adipokines and cytokines in ACC. Pro- inflammatory cytokines’ concentrations: TNFa, MCP1 and IL6 were higher in these compared to controls in our study. Due to the small sample size, results were not statistically significant and require further research. Some authors suggested a relationship between leptin levels and stimulation of proliferation of colon cancer cells [47]. It has also been indicated that leptin promotes proliferation of
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certain (not all) breast neoplasms in vitro, and promotes tumor invasiveness and angiogenesis in some animal models [47, 48]. It is also possible that leptin interacts with monocytes and macrophages, resulting in an increased production of pro-inflammatory cytokines including TNFa and IL6 [35].
In the current study there was a trend toward lower serum adiponectin concentrations in ACC patients compared to controls (not statistically significant). A small group of patients limits the findings of the study. In ACC patients compared to controls we only demonstrated statistically significant higher concentrations of IL6, TNFa and MCP1, i.e. cytokines with pro-inflammatory and proliferative effects. Higher concentrations of TNFa and IL6 may result in the reduction of adiponectin in this group of patients and indirectly affect tumor proliferation. Recent findings suggest leptin and adiponectin interact antagonistically on carcinogenesis, although this has not been clearly established in terms of cancer progression in vivo [48, 49]. ACC tumor size, and, consequently, resectability, is an important prognostic factor in patients with this neoplasm [33]. We found that leptin concentration was higher in ACC patients compared to controls. It is possible that higher leptin levels influence proliferation in ACC.
CONCLUSION
Adipokines and cytokines secreted by the adipose tissue influence the regulation of metabolism. Persistent adverse profile of adipocytokine secretion in SCS patients may contribute to increased long-term cardiovascular risk. Most adipokines and cytokines studied here (except for IL6) were not associated with higher secretion of glucocorticoids in the SCS group. A possible explanation of this result may be local adipocytokine secretion by periadrenal, perirenal and/or visceral fatty tissue or the adrenal gland itself. Further studies are required to elucidate the underlying pathomechanisms.
In the case of ACC patients, higher IL6, TNFa and MCP1 concentrations, i.e. cytokines with pro-inflammatory and proliferative effects, were recorded than in controls, and, an association between factors secreted by the adipose tissue and this cancer may be suggested.
AUTHORS’ CONTRIBUTIONS
AB contributed to planning and conducting the study, collecting and interpreting data, and drafting the manuscript. MK performed the statistical analysis. PK collecting data and preparing the manuscript.
KS approved the final draft submitted.
Compliance with Ethical Standards:
Funding: This research was supported with the funds of the Medical University of Gdansk grant number: ST-81.
Conflict of Interest: The authors declare that they have no conflicting interests
Ethical approval: The study was confirmed by the Independent Ethics Committee of Medical University of Gdansk (NKBBN/360-98/2016).
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Informed consent: Informed consent was obtained from all individual participants included in the study.
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| Variable | Cont (N=18) Mean ± SD or N (%) | SCS (N = 20) Mean ± SD or N (%) | ACC (N = 7) Mean ± SD or N(%) | p Cont vs SCS | p Cont vs ACC |
|---|---|---|---|---|---|
| Age [years] | 47.50±9.48 | 51.90±7.17 | 53.29± 13.49 | 0.120 | 0.328 |
| BMI [kg/m2] | 25.92 ± 3.28 | 27.92 ± 4.26 | 24.00 ± 2.83 | 0.111 | 0.171 |
| Female / male | 13 (72.2%)/5(27.8%) | 18(90.0%)/2(10.0%) | 6 (85.7%)/1(14.3%) | 0.166 | 0.637 |
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| Hypertension(HT)[%] | 0 (0.0%) | 5 (25.0%) * | 1 (14.3%) | 0.023 | 0.280 |
| DM [%] | 0 (0.0%) | 6 (30.0%) * | 0 (0.0%) | 0.011 | 1.000 |
| Fasting glucose [mg/dL] | 86.50± 11.86 | 107.95 ± 60.02 | 87.14± 13.45 | 0.143 | 0.914 |
| Fasting insulin [uU/mL] | 9.22±5.81 | 12.72 ±9.03 | 5.86± 3.31 | 0.169 | 0.173$ |
| HOMA index | 1.97± 1.29 | 3.95 ± 4.70 | 1.27 ±0.69 | 0.091 | 0.238$ |
| Systolic BP [mmHg] | 125.11 ±9.30 | 143.25 ±20.15* | 131.43 ± 12.15 | 0.001 | 0.246 |
| Diastolic BP [mmHg] | 71.11 ± 5.36 | 87± 11.29 * | 87.14 ±9.51 # | ☒ <0.001 | 0.003 |
| Total cholesterol [mg/dL] | 209.94 ± 34.99 | 200.84 ± 35.50 | 187± 16.11 # | 0.438 | 0.036 |
Legend: ACC - ACC patients; BP - blood pressure; Cont - healthy controls; SCS - SCS patients; * marks significantly differences between SCS patients and controls, # marks significant differences between ACC patients and controls; $ denotes p-value for U Mann-Whitney’s test (groups did not pass normality assumption).
| Variable | SCS Mean ± SD | Range in SCS patients | ACC Mean ± SD | Range in ACC patients | Normal range |
|---|---|---|---|---|---|
| Morning serum cortisol [nmol/L] | 404.75 ± 165.79 | 208-860 | 423.20 ± 225.20 | 217-805 | 101- 535 |
| Midnight serum cortisol [nmol/L] | 293.68 ± 185.00 | 102-900 | 133.40 ± 28.42 | 99-178 | 79-478 |
| Cortisol in 1 mg DXM test [nmol/L] | 247.95 ± 168.68 | 68-680 | 49.20 ± 21.10 | 36-86 | <50 |
| Morning ACTH [pg/mL] | 13.28 ± 8.10 | 10-45 | 51.43 ± 54.78 | 10-132 | 5-46 |
| UFC [nmol/24h] | 436.65 ± 304.35 | 94-1289 | 266.47 ± 93.14 | 179-364 | 12-486 |
| Androstendion [ng/ml] | 1.15±0.6 | 0.8-3.4 | 2.55± 0.8 | 1.2-3.5 | 0.7-3.6 |
| DHEAS [ug/L] | 34.09 ± 25.17 | 15-72.9 | 118.20 ± 109.15 | 48.7-244 | 42-290 |
Y
Legend: Cortisol in 1 mg DXM test - morning cortisol in the overnight 1 mg dexamethasone (DXM) suppression test; UFC - urinary free cortisol excretion; DHEAS - dehydroepiandrosterone sulphate.
| Variable | tests for independent samples (t test and alternative U Mann-Whitney's test ) | |||
|---|---|---|---|---|
| SCS patients | Controls | t | Normality U | |
ACCEPTED MANUSCRIPT
| Mean ± SD (N=20) | Mean ± SD (N=18) | (p-value) | Assumption | (p-value) | |
|---|---|---|---|---|---|
| Leptin [ng/ml] | 18.35± 13.89 * | 10.21 ±7.74 | 2.262 (0.031) | No | 1.856 (0.063) |
| MCP1 [pg/mL] | 360.17 ± 182.45 | 300.93 ± 97.04 | 1.267 (0.215) | No | 0.789 (0.430) |
| Resistin [ng/ml] | 12.43 ±5.01 | 10.77 ±5.24 | 0.994 (0.327) | Yes | 1.067 (0.286) |
| TNFa [pg/mL] | 0.99 ± 0.53 * | 0.68 ±0.40 | 2.062 (0.047) | No | 2.472 (0.013) |
| IL6 [pg/mL] | 2.47 ±2.67 | 1.39 ± 1.13 | 1.666 (0.108) | No | 2.193 (0.028) |
| Adiponectin [ng/ml] | 3271.10±1405.06 * | 6622.22 ± 4479.35 | -3.042 (0.006) | No | -3.290 (0.001) |
| Variable | tests for independent samples (t test and alternative U Mann-Whitney's test ) | ||||
|---|---|---|---|---|---|
| ACC patients Mean ± SD (N=7) | Controls Mean ± SD (N= 18) | t (p-value) | Normality Assumption | U (p-value) | |
| Leptin [ng/ml] | 5.68 ± 4.00 | 10.21 ±7.74 | -1.913 (0.070) | Yes | -1.242 (0.215) |
| MCP1 [pg/mL] | 549.37 ±234.38* | 300.93 ±97.04 | 2.715 (0.031) | No | 3.056 (0.002) |
| Resistin [ng/mL] | 12.22 ±4.90 | 10.77 ±5.24 | 0.652 (0.527) | Yes | 0.696 (0.486) |
| TNFa [pg/mL] | 1.54 ± 1.01 | 0.68 ± 0.40 | 2.183 (0.067) | No | 2.210 (0.027) |
| IL6 [pg/mL] | 8.33 ±4.37 * | 1.39 ± 1.13 | 4.151 (0.005) | No | 3.601 (<0.001) |
| Adiponectin [ng/mL] | 5833.71 ± 3253.50 | 6622.22 ± 4479.35 | -0.487 (0.634) | No | -0.393 (0.694) |
Highlights
· Adipokine and cytokine secretion in SCS may increase cardiovascular risk
· Adipokines and cytokines were not associated with secretion of glucocorticoids
· The association of adipose tissue with ACC may be suggested