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Evaluation of Vitamin-D, calcium, and phosphorus levels among diabetes mellitus type 2 in malwa belt of Punjab


1 Centre for Interdisciplinary Biomedical Research, Adesh University, Bathinda, Punjab, India
2 Centre for Interdisciplinary Biomedical Research, Adesh Institute of Medical Sciences and Research, Adesh University, Bathinda, Punjab, India
3 Department of Biochemistry, Adesh Institute of Medical Sciences and Research, Adesh University, Bathinda, Punjab, India
4 Department of Biochemistry, Guru Gobind Singh Medical College, BFUHS, Faridkot, Punjab, India

Date of Submission21-May-2020
Date of Decision19-Jun-2020
Date of Acceptance03-Jul-2020

Correspondence Address:
Saranpal Singh,
Department of Biochemistry, Adesh Institute of Medical Sciences and Research, Adesh University, Bathinda, Punjab
India
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/mjdrdypu.mjdrdypu_227_20

  Abstract 


Background: Type-2 diabetes mellitus (T2DM) or noninsulin-dependent diabetes is an endocrinological disease of impaired metabolism of carbohydrates. T2DM is increasing both nationally and worldwide at an alarming rate. In intermediary metabolism, including enzyme activities, electrical gradients and cellular function Vitamin D, calcium and phosphorus play an important role. Disturbances in the levels of Vitamin D, calcium, and phosphorus were found to be associated with T2DM. The present study is aimed to evaluate the levels of Vitamin D, calcium, and phosphorus levels in 300 patients, of which 150 were nondiabetic healthy individuals (control) and 150 were T2DM patients. Materials and Methods: A volume of 5 ml of the blood sample was taken after 12 h of overnight fasting by complete aseptic means from the antecubital vein with the sterilized disposable syringe for fasting plasma glucose (FPG), postprandial plasma glucose (PPPG), glycated hemoglobin, serum 25 hydroxy (OH) Vitamin D (25[OH]D), calcium, and phosphorus. Results: Of 150 type-2 diabetic and 150 nondiabetic group, there was a significant statistical difference between the diabetic and nondiabetic group for FPG (P < 0.001), PPPG (P < 0.001), 25(OH) D (P < 0.001), calcium (P < 0.001), and phosphorus (P < 0.001). 25(OH) D, calcium, and phosphorus levels were decreased in T2DM patients as compared to the control group. Conclusion: This study revealed that there is a significant decrease in the levels of Vitamin D, calcium, and phosphorus in Type-2 diabetic patients, which may be an important factor in the early onset of the disease in susceptible individuals.

Keywords: 25 hydroxy Vitamin D, calcium and phosphorus, Type-2 diabetes mellitus



How to cite this URL:
Nigah SL, Jagota G, Singh S, Goyal G. Evaluation of Vitamin-D, calcium, and phosphorus levels among diabetes mellitus type 2 in malwa belt of Punjab. Med J DY Patil Vidyapeeth [Epub ahead of print] [cited 2021 Dec 6]. Available from: https://www.mjdrdypv.org/preprintarticle.asp?id=321338




  Introduction Top


The incidence of diabetes mellitus Type 2 (T2DM) is increasing at an alarming rate; the significant increase in the number of people with diabetes will be in India.[1] Most of the expected population growth between 2000 and 2030 will be concentrated in the urban areas of the world.[1] The total number of people with diabetes is projected to rise from 171 million in 2000 to 366 million in 2030.[2] In developing countries, the majority of people with diabetes are in the age group of 45–64 years, in contrast: the majority of people with diabetes in developed countries are >64 years of age.[3] By 2030, it is estimated that the number of people with diabetes >64 years of age will be >82 million in developing countries and >48 million in developed countries.[3]

Diabetes mellitus 2 is the endocrine disease characterized by hyperglycemia due to progressive failure of pancreatic cells in a setting of chronic insulin resistance.[4] In experimental studies, calcium and Vitamin D have been shown to improve pancreatic beta-cell function and peripheral insulin sensitivity, whereas low Vitamin D status is associated with markers of impaired glucose metabolism[4],[5] and insulin resistance.[6] In recent decades, insulin resistance and secretion have been shown to depend on calcium and Vitamin D homeostasis.[7] Insulin secretion is a calcium-dependent process requiring the influx of calcium to the beta-cell.[7],[8]

The phosphate lowering action of insulin was recognized immediately after insulin preparations became available.[9] There is a close correlation between the plasma concentration of phosphorus and the degree of metabolic control of diabetes.[10] Various studies have shown great interest in low phosphorus diet on glucose homeostases such as insulin resistance and glucose tolerance.[11] The main question is whether diabetes state had an impact on Vitamin D, calcium, and phosphorus. Therefore, the main aim of this study is to assess, is there any differences of serum Vitamin D, serum calcium and serum phosphorus value of diabetics versus nondiabetic persons and to find out the correlation of serum Vitamin D, serum calcium, and serum phosphorus with some biochemical parameters in two groups.


  Materials and Methods Top


Study group (inclusion criteria)

The present study was a comparative case–control study conducted at a tertiary care hospital from March 2018 to March 2019 in the Department of Biochemistry, Guru Gobind Singh Medical College and Hospital, Baba Farid University of Health Sciences Faridkot, Punjab in collaboration with Department of Medicine, Guru Gobind Singh Medical College and Hospital, Baba Farid University of Health Sciences Faridkot, Punjab. The minimum sample size for each group has been calculated using the formula: N = (1 + 1/k) [σ(Z1−α/2 + Z1−β) 2/(ua − ub)].[12] Assuming equal group sizes to achieve a power of 80% and a (two sided) confidence level of 90%, the study requires a sample size of 99 for each group. Therefore, 150 diagnosed cases of T2DM of age group 30–60 years and 150 age- and sex-matched healthy individuals were recruited as a control group in the study. This study was conducted after taking the institutional approval by the ethical committee of Guru Gobind Singh Medical College and Hospital, Baba Farid University of Health Sciences Faridkot, Punjab (Ref. no. GGS/IEC/18/82, dated February 15, 2018) and after obtaining written consent from all the patients.

Exclusion criteria

Patients of T2DM with other complications such as chronic liver disease, chronic kidney disease, or patient taking drugs affecting Vitamin D metabolisms such as rifampicin, isoniazid, ketoconazole, and phenytoin or patients taking Vitamin D therapy were excluded from the study. Pregnant women, type-1 diabetes mellitus, and hypertension patients were also excluded from the study.

Laboratory tests

A sample of 5 ml venous blood was collected in both fasting and postprandial state under aseptic precautions out of it 3 ml of blood was poured in plan sterilized vial and allowed it to clot at room temperature for half an hour. Serum was separated by centrifugation process at 3000 revolutions per minute for 10 min, and it was utilized for the estimation of serum 25-hydroxy Vitamin D (25[OH] D), serum calcium, and serum phosphorus.

For the investigation of fasting plasma glucose (FPG), 2 ml of blood was poured in sugar vial containing an anticoagulant (sodium fluoride and potassium oxalate). In addition, postprandial plasma glucose (PPPG) was measured, 2 h after meals (75 g of glucose in a glass of water) by using standard kits. Serum 25(OH) D was estimated with the ELISA method.[13] Estimation of serum calcium was done with Arsenazo 111 method/cresalphthalein complexions method.[14] Estimation of serum phosphorus with Molybdate U. V. method.[15] FPG and 2-h PPPG were measured using enzymatic methods (GOD-POD).[16] Weight and height were measured, and body mass index (BMI) was calculated by dividing weight (kg) by the square of height (m). The study was conducted after the institutional approval by the ethical committee of Guru Gobind Singh Medical College and Hospital, Baba Farid University of Health Sciences Faridkot, Punjab (Ref. No. GGS/IEC/18/82 dated February 15, 2018) after obtaining written consent from all the patients.

Statistical analysis

Data were expressed in terms of mean ± standard deviation (SD). The independent t-test was used to determine the significance of any baseline differences between groups and Pearson correlation was calculated using the expression. The data were analyzed with excel software version 12. Values of P < 0.05 were assumed to be significant (P < 0.05).


  Results Top


This was a case–control study conducted on 150 cases of T2DM (n = 150) and 150 age- and sex-matched healthy controls (n = 100) [Table 1] provides a summary of the demographic characteristics of the study population (n = 300). The majority of the study population were female (56.0%) and belonged in the age group of 51–60 years. Results showed no significant difference in sex, age, area, and BMI between nondiabetic healthy individuals and T2DM patients. The results were expressed as mean ± SD. There was a statistically highly significant difference (P < 0.001) in age, BMI, FPG, PPPG, glycated hemoglobin (HbA1c), serum Vitamin D, calcium, and phosphorus levels [Table 2]. Results showed statistically significant difference (P < 0.05) in BMI, and nonsignificant difference (P > 0.05) in age, FPG, PPPG, HbA1c, serum Vitamin D, calcium and phosphorus levels between Type-2 diabetic male and Type-2 diabetic female [Table 3]. The levels of serum Vitamin D, serum calcium, and serum phosphorus were nonsignificantly lower, and levels of age, BMI, FPG, PPPG, and HbA1c were nonsignificantly higher in Type-2 diabetic female than in Type-2 diabetic male [Table 3].
Table 1: Demographic Information of nondiabetic healthy individuals (control) and Type-2 diabetic mellitus patients (cases)

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Table 2: Comparison of age, fasting plasma glucose, postprandial plasma glucose, serum Vitamin D, serum calcium and serum phosphorus in nondiabetic healthy individuals and Type-2 diabetic mellitus patients

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Table 3: Comparison of age, fasting plasma glucose, postprandial plasma glucose, serum Vitamin D, serum calcium, and serum phosphorus in Type 2 diabetic individuals

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There was a positive correlation between age, BMI, FPG, PPPG, and HbA1c, and age, BMI, FPG, PPPG, HbA1c was negatively correlated with serum Vitamin D, calcium and phosphorus levels [Table 4]. Results of the present study showed an increased deficiency of serum levels of Vitamin D, calcium, and phosphorus in Type-2 diabetic females than in Type-2 diabetic males, but the difference was statistically nonsignificant (P < 0.05) [Table 5]. The study also showed an increased deficiency of serum levels of Vitamin D, calcium and phosphorus in normal healthy females than in normal healthy males, but the difference was statistically nonsignificant (P < 0.05) [Table 6].
Table 4: Correlation of fasting plasma glucose, postprandial plasma glucose, serum Vitamin D, serum calcium, and serum phosphorus in Type-2 diabetic individuals and normal healthy individuals

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Table 5: Variation in serum Vitamin D, serum calcium, serum phosphorus levels among Type-2 diabetic individuals

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Table 6: Variation in serum Vitamin D, serum calcium, serum phosphorus levels among normal healthy individuals

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Comparison of FPG, PPPG, HbA1c, S. VIT D, S. CAL, S. PHOS levels of diabetic individuals distributed according to BMI was carried out, and there was a significant difference in FPG, PPPG, HbA1c, S. PHOS levels and nonsignificant difference in S. VIT D, S. CAL levels [Table 7]. The present study showed the comparison of FPG, PPPG, HbA1c, S. CAL, S. PHOS levels of male and female diabetic individuals distributed according to S. VIT D levels and there was a significant difference in FPG, PPPG, HbA1c, S. VIT D, S. CAL levels and nonsignificant difference in S. PHOS levels [Table 8] and [Table 9].
Table 7: Comparison of fasting plasma glucose, postprandial plasma glucose, glycated hemoglobin, serum Vitamin D, serum calcium, serum phosphorus levels of diabetic individuals distributed according to body mass index

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Table 8: Comparison of fasting plasma glucose, postprandial plasma glucose, glycated hemoglobin, serum calcium, serum phosphorus levels of female diabetic individuals distributed according to serum Vitamin D levels

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Table 9: Comparison of fasting plasma glucose, postprandial plasma glucose, glycated hemoglobin, serum calcium, serum phosphorus levels of male diabetic individuals distributed according to serum Vitamin D levels

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  Discussion Top


The present study showed that the prevalence of T2DM was more frequent in females 56.0% than males 44.0%; this finding agreed with a previous study done by Marwa et al. in which prevalence of T2DM in females was 63% than males, i.e., 37%.[17] Furthermore, the present study showed that obesity was more common in T2DM females 28.6% than in T2DM males 25.7% this result agreed with a previous study done by McCord et al. and Marwa et al. who stated that the obesity effect women more than men.[17],[18] The present study showed that Vitamin D deficiency was more frequent in T2DM females 79.7% than males 71.2% this finding confirmed by Arif et al. 2017 who reported that T2DM female were four times more prone for Vitamin D deficiency than males 151 (79%) and 40 (21%), respectively.[19] Vitamin D levels of patients were found to be in the range of 2.25–55.13 while that of controls were in the range of 1.51–71.0 ng/ml which slightly varies from a study conducted by Bayani et al., in which mean concentration of Vitamin D in the case group was 18.7 ± 10.2 and in the control group was 24.6 ± 13.5 ng/dl.[12] The results showed that Vitamin D concentration was significantly lower in Type-2 diabetic patients than the healthy individuals, which was supported by several workers in the previous studies by Pittas et al. 2010, Anderson et al. 2010, and Knekt et al. 2008.[20],[21],[22]

Another important finding was the significant negative association of Vitamin D level with age in diabetic patients (r = −0.145, P = 0.001), which was consistent with the study by Hagenau et al. 2009.[23] Hagenau et al. 2009 showed, serum 25(OH) D levels varied with age.[23] Serum calcium levels in patients were found to be in the range of 7.20–11.40 mg/dl, while the range of serum calcium concentration in the healthy individuals under study was 7.0–10.80 mg/dl. Thus, serum calcium concentration was significantly lower in patients as compared to controls. In this study, there was a significant difference in serum Vitamin D, calcium, and phosphorus levels between Type-2 diabetic and nondiabetic individuals.

Serum phosphorus levels in patients were found to be in the range of 2.8–6.0 mg/dl, whereas the range of serum phosphorus concentration in the healthy individuals understudy was 2.4–10.80 mg/dl. Thus, serum phosphorus concentration was also comparatively lower in patients as compared to controls with P value 0.001 at (P < 0.05). Serum phosphorus levels were higher in both male diabetic patients as well as in normal male healthy controls under study (P < 001) with higher serum Vitamin D levels but no difference in the case of female patients. Our results were the same as studies of Raab et al., Al-Shoumer et al., and Bierschenk et al., which revealed T2DM patients as compared to nondiabetic individuals have lower serum Vitamin D concentrations.[24],[25],[26] In this study, the mean of Vitamin D in the individual with and without diabetes showed the presence of Vitamin D deficiency in both groups. However, differences in results as compared to other studies may be related partly to the high prevalence of Vitamin D deficiency in both groups. Several factors potentially influence on Vitamin D status, such as adiposity, genetic factors, and issues that have an effect on the cutaneous synthesis of Vitamin D such as season, skin pigmentation, melanin concentration, age, clothes, and consumption sunscreens Gaafar and Badr 2013.[27] It seems that these results are due to imperfect exposure to sunlight, and little seafood ingestion possibly influenced Vitamin D status in these groups. The change in Vitamin D status with age probably might be the result of a lack of sunlight exposure related to social factors, supplementation intake, and physical inactivity.

The study concluded that there was a significant difference in serum 25(OH) D, calcium, and phosphorus levels between Type-2 diabetic patients and the control group. There appears to be a relationship between Vitamin D, phosphorus, and calcium status in T2DM. This study suggests that deficiency of Vitamin D influences postprandial glycemia and insulin response, as Vitamin D is needed for the release of insulin from the beta cells of islet of Langerhans of the pancreas it may be playing a role in the development of diabetes.


  Conclusion Top


The decreasing levels of serum Vitamin D, calcium, and phosphorus may influence glycemia. The assessment of serum Vitamin D, calcium, and phosphorus levels are important to monitor the prognosis of T2DM. As Vitamin D, calcium and phosphorus levels were inversely associated with the development of T2DM and maybe with combined nutrients, glucose metabolism can be beneficial.

Acknowledgments

The help provided by the Chairperson, Dr. Ram Gopal Saini, Centre for Interdisciplinary Biomedical Research, Adesh University, is gratefully acknowledged.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6], [Table 7], [Table 8], [Table 9]



 

 
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