|Year : 2022 | Volume
| Issue : 1 | Page : 17-20
Evaluating the Interrelationship Between Salivary Cortisol and Glycemia in Type 2 Diabetics
Tharani Vijaya Kumar1, Vinoth Vasanthi1, Thayalan Dinesh Kumar1, Madhu Narayan1, Bose Divya1, Raj Kumar Krishnan1, Ramya Ramadoss2
1 Department of Oral Pathology & Microbiology, SRM Dental College, SRMIST, Ramapuram, Chennai, India
2 Department of Oral Biology, Saveetha Dental College, Chennai, India
|Date of Submission||30-May-2022|
|Date of Decision||07-May-2022|
|Date of Acceptance||11-May-2022|
|Date of Web Publication||05-Aug-2022|
Dr. Tharani Vijaya Kumar
Department of Oral Pathology & Microbiology, SRM Dental College, Bharathi Salai, Ramapuram, Chennai-600089, Tamil Nadu
Source of Support: None, Conflict of Interest: None
Introduction: Diabetes mellitus is a multifactorial metabolic condition marked by high blood sugar levels as well as problems with carbohydrate, lipid, and protein metabolism. Impaired insulin secretion and hepatic gluconeogenesis cause high blood sugar levels. Cortisol in the saliva is the unbound, free hormone, with absolute quantities between 30% and 50% fewer than in blood. The aim of the study was to evaluate the levels of salivary cortisol in normal healthy controls and type 2 diabetics. The objective was to evaluate the morning and evening levels of salivary cortisol in normal and type 2 diabetics by enzyme-linked immunosorbent assay (ELISA). Materials and Methods: 20 patients in the age group of 30 to 60 years with type 2 diabetes with HbA1c <6.5% were included in Group I. Twenty participants in the age group of 30 to 60 years were included in Group II. Salivary samples were collected and centrifuged at –4°C at 3500 rpm and ELISA was performed. Data were analyzed using Statistical Package for the Social Sciences version software version 16. Paired sample t test was used. Results: The mean morning salivary cortisol level of diabetic patients (Group I) was 1.62 mmol/dL and the evening was 1.34 mmol/dL. The difference was statistically significant (P = 0.00001). The mean salivary cortisol level of healthy subjects in the morning was 0.47 mmol/dL and evening was 0.29 mmol/dL, which is statistically significant (P = 0.00001). Conclusion: From this study, we can conclude that the salivary cortisol level is increased in patients with type 2 diabetes when compared to healthy individuals. There are no obvious changes in morning and evening salivary cortisol levels of diabetes patients.
Keywords: Enzyme-linked immunosorbent assay, glucocorticoid, oral manifestations, salivary cortisol, type 2 diabetics
|How to cite this article:|
Kumar TV, Vasanthi V, Kumar TD, Narayan M, Divya B, Krishnan RK, Ramadoss R. Evaluating the Interrelationship Between Salivary Cortisol and Glycemia in Type 2 Diabetics. J Orofac Sci 2022;14:17-20
|How to cite this URL:|
Kumar TV, Vasanthi V, Kumar TD, Narayan M, Divya B, Krishnan RK, Ramadoss R. Evaluating the Interrelationship Between Salivary Cortisol and Glycemia in Type 2 Diabetics. J Orofac Sci [serial online] 2022 [cited 2022 Oct 2];14:17-20. Available from: https://www.jofs.in/text.asp?2022/14/1/17/353478
| Introduction|| |
Diabetes mellitus (DM) is a multifactorial metabolic condition marked by high blood sugar levels as well as a disturbance in carbohydrate, lipid, and protein metabolism. Impaired insulin secretion and hepatic gluconeogenesis cause high blood sugar levels. Chronic hyperglycemia causes various complications to systemic health, as well as oral health, so control of hyperglycemia is very important. Impaired neutrophil function, increased collagenase activity, decreased collagen synthesis, microvascular disease, and neuropathy are possible pathways that may be involved in oral consequences of diabetes. Oral manifestations associated with DM include dry mouth (xerostomia), dental caries (including root caries), periapical lesions, gingivitis, periodontal disease, oral candidiasis, burning mouth, glossodynia, altered taste, oral lichen planus, recurrent aphthous stomatitis, increased infection, and poor wound healing.
Cortisol in the saliva is free and unbound with absolute quantities between 30% and 50% fewer than in blood. The pathophysiology of mood, anxiety, cognitive, and other diseases has been linked to the stress-responsive hypothalamic pituitary adrenal (HPA) axis. The axis consists of stimulating forward and feedback inhibition loops involving the brain, pituitary, and adrenal glands to produce glucocorticoid end products. Glucocorticoids play a role in the regulation of mineralocorticoids, immune system function, blood pressure, and metabolism. Obesity and glucose intolerance, hypertension, hypercholesterolemia, and other diseases are related to elevated cortisol levels. Patients with uncontrolled diabetes experienced diurnal glucocorticoid hypersecretion and changes in the regulation of the HPA axis. In diabetes patients, the progressive increase in cortisol level will further worsen the disease process. The present study aimed to address the difference in salivary cortisol levels among type 2 diabetics and healthy age-matched individuals. The present study is different from the studies reported in the literature as the HbA1c levels were also correlated with the cortisol levels among the type 2 diabetics.
| Materials and Methods|| |
Ethical approval for this study ((SRMU/M&HS/SRMDC/2021/PG/012) was provided from, SRM dental college and hospital SRM University, Chennai, on 20 jan 2021. Study participants were recruited from the outpatient department of the Institution after obtaining informed consent. Age and sex matched healthy individuals and type 2 diabetics were included in the study. Twenty patients aged 30 to 60 years with type 2 diabetes and HbA1c >6.5% were included in Group I. Twenty healthy individuals aged 30 to 60 years were included in group II. Patients with any other systemic illness, psychological illness, obese individuals, antidepressant medication, corticosteroids, or history of smoking, alcohol, and chewing were excluded from the study.
Two milliliters of unstimulated saliva was collected from patients belonging to both groups. Proper instructions were given prior to sample collection, such as avoidance of acidic fluids, and solid foods half an hour prior to sample collection. Study participants were made to sit erect and expectorate 2 mL of unstimulated saliva into sterile containers. Samples were collected in the morning (8–10 am) and evening (3–5 pm) as well to avoid diurnal variation. Following collection, saliva was immediately centrifuged at 2000 rpm for 10 minutes. The resulting supernatant was separated into 1 mL of aliquots and stored at –20°C until further processing.
Sandwich enzyme-linked immunosorbent assay kit was used for the quantitative detection of salivary cortisol. The plate was pre-coated with a human cortisol antibody. All reagents, standard solutions, and samples were brought to room temperature before use. 50 μL of the standard was pipetted to the standard well, then 40 μL of the sample was added to sample wells, and finally 10 μl of anti-cortisol antibody was added to sample wells. Following this, 50 μL of streptavidin-horseradish peroxidase was added to sample wells and standard wells. The plate was kept covered with a sealer and incubated at 37°C for 60 minutes. Following this, the sealer was removed, the plate was washed with wash buffer, and wells were soaked with 0.35 mL of wash buffer for 30 seconds to 1 minute for each wash. 50 μL of substrate solution A was added to each well and then 50 μL of substrate B was added to each well and the plate was kept covered in the dark. The color changed to yellow with the addition of 50 μL of stop solution to each well. Optical density was determined within 10 minutes of the addition of the stop solution with a microplate reader set to 450 nm.
Data were analyzed using Statistical Package for the Social Sciences version software version 16. Paired sample t test was used to compare the diurnal variation between morning and afternoon samples within each group. P less than 0.05 was considered significant.
| Results|| |
In this study, 20 diabetic patients and 20 healthy people participated. Out of 40 people, 10% of males and 12.5% of females were in the age group of 31 to 40 years, 22.5% of males and 20% of females were in the age group of 41 to 50 years, and 22.5% of males and 12.5% of females were between 51 and 60 years [Figure 1]. The mean morning salivary cortisol level of diabetic patients (Group I) was 1.62 mmol/dL and the evening was 1.34 mmol/dL [Figure 2]. The difference was statistically significant (P = 0.00001). The mean salivary cortisol level of healthy subjects in the morning was 0.47 mmol/dL and evening was 0.29 mmol/dL [Figure 2]. The difference was statistically significant (P = 0.00001). A significant difference in mean salivary cortisol levels was found in comparing the morning and evening levels between Group I and Group II (P = 0.00001). The mean salivary cortisol levels were also found to increase with the increase in HbA1c levels among the type 2 diabetic patients [Table 1].
|Figure 2 Mean morning and evening salivary cortisol levels – Group I and Group II|
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|Table 1 Distribution of mean salivary cortisol level among type 2 diabetics with different HbA1c range|
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| Discussion|| |
DM is derived from the Greek word diabetes, which means “to pass through,” and mellitus, which means “sweet.” Insulin resistance can be caused by a variety of factors, but it is most commonly associated with obesity. Type 2 DM is characterized by a more complex interaction of genetics and lifestyle. Insulin deficiency from injury to islet beta cells affects glucose utilization by skeletal muscle, liver, and adipose tissues.
According to a study conducted by Shirzaii et al. in 2016, the mean salivary cortisol level in type 2 diabetic patients was 1.73, while it was 1.08 in healthy subjects, indicating that the mean cortisol level in diabetic patients was higher than healthy subjects. The present study was also concordant with the same results. The mean salivary cortisol levels in type 2 diabetic patient were higher than in healthy individuals.
A study by Chiodini et al. in 2007 reported that hypothalamus-pituitary-adrenal axis activity and cortisol secretion in diabetic patients were higher than in healthy subjects. HbA1c was also reported to affect the cortisol levels in this study. The current study also reveals that the mean salivary cortisol level in type 2 diabetic patients was higher as compared to healthy subjects. HbA1c of the diabetic group was also found to affect the cortisol levels in the present study.
In 2014, Hackett et al. documented that salivary cortisol levels in type 2 diabetics increased only during sleep and slightly changed during the day, whereas the present study revealed that morning levels were high. The difference in the salivary cortisol levels might be due to the diurnal variation.
In 2004, Radahmadi et al. stated that cortisol secretion levels in people who have diabetes increased significantly and the effects of diabetes on cortisol secretion are higher than the effects of mental stress. DM is a powerful stimulant to the body’s physiological system. Chiodini et al. in 2007 evaluated HPA activity in type 2 diabetic subjects with and without chronic complications and nondiabetic patients. The study documented that cortisol level was increased in diabetic subjects with chronic complications. According to cross-sectional research on the relationship between diurnal cortisol and diabetes, patients with type 2 diabetes had a flatter diurnal cortisol curve than persons without diabetes. This is in concordance with the observations of our study with not much of a difference between morning and evening salivary cortisol levels in type 2 diabetics.
| Conclusion|| |
From this study, we can conclude that only the salivary cortisol level is increased in type 2 diabetics when compared to healthy individuals. This present study found that increase in cortisol levels in diabetics might be due to mental stress, hormonal imbalance, and various other factors. Future research with an increase in sample size is recommended to delineate the association between hyperglycemia and cortisol levels.
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Conflicts of interest
There are no conflicts of interest.
| References|| |
Salehi M, Mesgarani A, Karimipour S, Pasha SZ, Kashi Z, Abedian S, Mousazadeh M, Molania T, Comparison of salivary cortisol level in type 2 diabetic patients and pre-diabetics with healthy people. Open Access Macedonian Journal of Medical Sciences 2019;7:2321.
Cicmil A, Govedarica O, Lečić J, Mališ S, Cicmil S, Čakić S. Oral symptoms and mucosal lesions in patients with diabetes mellitus type 2. Balk J Dent Med 2017;21:50–4.
Rohani B. Oral manifestations in patients with diabetes mellitus. World Journal of Diabetes 2019;10:485.
Rosmond R. Stress induced disturbances of the HPA axis: a pathway to type 2 diabetes? Med Sci Mon 2003;9:RA35–9.
Schatzberg AF, Keller J, Tennakoon L et al.
, HPA axis genetic variation, cortisol and psychosis in major depression. Mol Psychiatry 2014;19:220–7.
Chan O, Inouye K, Riddell MC, Vranic M, Matthews SG. Diabetes and the hypothalamo-pituitary-adrenal (HPA) axis. Minerva Endocrinol 2003;28:87–102.
Rajaei E, Jalali MT, Shahrabi S, Asnafi AA, Pezeshki SMS. HLAs in autoimmune diseases: dependable diagnostic biomarkers? Curr Rheumatol Rev 2019;15:269–76.
Stumvoll M, Goldstein BJ, Van Haeften TW. Type 2 diabetes: principles of pathogenesis and therapy. The Lancet 2005;365:1333–46.
Shirzaii M, Heidari F, Moradi V. Comparison of salivary cortisol level in patients with controlled type II diabetes and healthy subjects. J Qazvin Univ Med Sci 2016;20:34–40.
Chiodini I, Adda G, Scillitani A et al.
, Cortisol secretion in patients with type 2 diabetes: relationship with chronic complications. Diabetes Care 2007;30:83–8.
Hackett RA, Steptoe A, Kumari M. Association of diurnal patterns in salivary cortisol with type 2 diabetes in the Whitehall II study. J Clin Endocrinol Metab 2014;99:4625–31.
Radahmadi M, Shadan T, Sadr SS, Karimian S. The effect of psychical stress on cause and exacerbation of diabetes mellitus, serum glucose and cortisol levels, and body weight in rats. J Shahrekord Univ Med Sci 2004;6:14–25.
Chiodini I, Adda G, Scillitani A, Coletti F, Morelli V, Di Lembo S, Epaminonda P, Masserini B, Beck-Peccoz P, Orsi E, Ambrosi B. Cortisol secretion in patients with type 2 diabetes: relationship with chronic complications. Diabetes care 2007;30:83–8.
Joseph JJ, Golden SH. Cortisol dysregulation: the bidirectional link between stress, depression, and type 2 diabetes mellitus. Annals of the New York Academy of Sciences 2017;1391:20–34.
[Figure 1], [Figure 2]