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Volume 14, Issue 3 (2022)
3 2022, 14(3): 317-321 |
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Received: 2022/05/18 | Accepted: 2022/08/1 | Published: 2022/08/15
Received: 2022/05/18 | Accepted: 2022/08/1 | Published: 2022/08/15
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Mohammad A, Hussein H, Kadhim S, Hashoosh M, Hassan E. A Comparative Study of Lead and Cadmium Concentrations in Serum Samples of Diabetic Patients in Al-Najaf Governorate. 3 2022; 14 (3) : 10
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1- Department of Pharmacology & Therapeutics, Faculty of Medicine, University of Kufa, Najaf, Iraq
2- Physical Planning Unit, Middle Euphrates Oncology Center, Najaf, Iraq
3- Department of Physics, Faculty of Sciences, University of Kufa, Najaf, Iraq
4- The Ministry of Education, Baghdad, Iraq
2- Physical Planning Unit, Middle Euphrates Oncology Center, Najaf, Iraq
3- Department of Physics, Faculty of Sciences, University of Kufa, Najaf, Iraq
4- The Ministry of Education, Baghdad, Iraq
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Introduction
Diabetes mellitus (DM) is a group of metabolic disorders characterized by high blood sugar [1]. It is a chronic disease that occurs when the body is not able to use insulin effectively or when the pancreas does not excrete enough insulin [2]. DM is a global disease that spreads all over the world at a rate that differs from country to country [3]. This disease is usually divided into two types, are the most common, type1 (destruction of β cells with an absolute lack of insulin), and the percentage of people with this type is 5-10%, while the second type is the most prevalent 90-95% of cases, the subjects suffer from a relative lack of insulin secretion [4]. Minerals and trace elements are essential micronutrients required for the normal functioning body. These elements are particularly beneficial for physiological functions [5]. A relationship has been reported between trace elements and diabetes mellitus [6]. Minerals and trace elements are essential for many biochemical reactions, and present as stabilizing components of enzymes and proteins in addition they function as cofactors for many enzymes. Certain trace elements regulate crucial biological processes by binding to the receptor site of the cell membrane or by changing the shape of the receptor to prevent the entry of particular molecules into the cell [7]. Some essential trace metals including copper (Cu), zinc (Zn), magnesium (Mg), iron (Fe), and calcium (ca) are associated with diabetes mellitus type 2 [8]. There are many trace elements altered in diabetes mellitus and might have specific roles in the pathogenesis and progress of this disease, they reported higher levels in the blood, urine, and scalp hair of diabetics than nondiabetics [9]. Lead (Pb) is harmful to most of the human body organs and obstructs metabolism and cellular activities [10]. A linear relationship is reported between blood Pb level and renal dysfunction in age-related diseases after recurrent exposures to Pb [11]. It badly affects the antioxidant pathways [12]. The metal-induced toxicity may cause derangement of antioxidant mechanisms in living tissues; as a result, highly reactive oxygen species (ROS) are produced. This antioxidant discrepancy might lead to the degradation of proteins, nucleic acids, and lipid peroxidation. An oxidative attack of cellular components by ROS is concerned with the pathogenesis of numerous human diseases including diabetes [13].
Cadmium (Cd) is a heavy metal that is broadly detected in air, water, and soil. Increased Cd level in water is absorbed by plants, animals, and humans [14]. Frequent exposure to Cd supplied to its excess addition in the kidney causes renal damage and nephropathy [15]. The high level of Cd reduces calcium absorption, which becomes an approaching cause of bone and kidney losses, called Itai-Itai disease. The Cd might down-regulate glucose transporter-4 (GLUT4) translocation by insulin [16].
A previous study proved that blood Cd and Pb levels were higher in diabetics with reduced antioxidant concentration, and it suggested that the elevation of toxic metals may be a contributing factor to the pathogenesis of diabetes mellitus [17].
These studies have been done in some areas of the world, but these studies in Iraq are very few and this is the first study in the Al-Hira area. Therefore, this study aimed to investigate the concentration of these two elements in different serum samples for diabetic patients and compared them with healthy persons.
Materials and Methods
This pilot study was conducted in the Al-Hira area, Najaf governorate, Iraq from June 2021 to January 2022. It included 25 diabetic patients (12 males and 13 females) and 15 healthy non- diabetic subjects (7 males and 8 females) whose ages ranged between 30-80 years. The diabetic patients with an average age of 52.52 years, have type 1 or type 2 diabetes. Healthy people have an average age of 53 years.
The blood samples were taken after fasting one night and centrifuged after acid digestion to obtain serum which was stored for later use. Lead and Cadmium measurements were done by using an atomic absorption spectrophotometer, SHIMADZU model AA-7000 atomic absorption spectrophotometer (AAS) made in Japan which was calibrated to cadmium and lead elements to obtain values controlled for the groups under study as in Diagrams 1 & 2.
Diagram 1) Calibration of the atomic absorption system for cadmium
Diagram 2) Calibration of the atomic absorption system for lead
Microsoft Excel 2019 was used for spreadsheets, SPSS software version 20 was used to analyze the data, and used the independent t-test and Pearson Correlation to find the statistical significance between the different samples which depend on p<0.05.
Findings
The study population included 25 diabetic patients (12 males and 13 females) and 15 healthy non- diabetic subjects (7 males and 8 females) so the gender is comparable in number between case and control groups. Age ranged between 30-80 years. The diabetic patients had an average age of 52.52 years and the healthy people had 53 years’ average age with a non-significant difference between the two groups. All the participants were living in the Al-Hira area, Najaf governorate in Iraq, and this was the first study for these heavy elements in that area.
This study demonstrated that the cadmium concentration in diabetic patients was higher than in the control group with a high statistical significance (p<0.0001), while the lead concentration is significantly lower in the diabetic group as compared to the healthy group (Table 1). According to our results, the cadmium concentration in diabetic male patients was significantly higher (p<0.001) than in the male of the control group, meanwhile lead concentration in the serum of diabetics was insignificantly (p>0.05) lower than in healthy male persons (Table 1; Diagram 4). The same results were found when comparing concentrations of Cd and Pb in the serum of the female diabetic and control group (Table 1).
Table 1) Concentrations of Cd and Pb elements in the serum of healthy and diabetic patients
Pearson correlation analysis was carried out to determine the relationship between variables (Table 2). There was a strong inverse relationship between cadmium and lead in healthy people, with a statistically significant correlation. There was also a strong inverse and statistically significant association between cadmium in healthy subjects with cadmium in people with diabetes, and a moderate positive correlation between cadmium in healthy people and lead in diabetic patients without statistical significance, and finally, the correlation was negative and not statistically significant between lead in healthy people and cadmium in diabetic patients with the negligible correlation between cadmium among sick and healthy subjects.
Table 2) Comparative study for Pearson correlation r between (Cd and Pb)
In Table 3, there was no statistically significant between the different groups of the studied serum samples for the lead trace. There was statistically significant between healthy versus type 1 and healthy versus type 2 Groups using the independent test as well as for the triple groups (control, type 1, type 2) using the ANOVA test (p<0.05).
Table 3) Comparison of serum samples between healthy and types 1 and 2 diabetes groups
Diagram 3) Illustration of Cadmium and lead serum levels in healthy, type1, and type 2 subjects
Discussion
According to the studied samples that we have in our study, cadmium concentration (for both males and females) in diabetic patients is higher than the healthy ones These results were in agreement with other clinical research which found that Cd level is increased in diabetic and prediabetes patients [17-20].
A cohort study in China suggested that high chronic Cd exposure for general population adults might contribute to elevated fasting blood glucose and result in type 2 diabetes mellitus development [21].
Both experimental and epidemiological studies showed that exposure to the environmental pollutant cadmium is associated with hyperglycemia, type 2 diabetes, and reduced serum insulin. Individual cellular effects of cadmium are reviewed, it is likely that no single mechanism is involved, somewhat multiple mechanisms exist, and work synergistically ensuing in islet dysfunction and finally dysglycemia [22].
it was found that Cadmium, a non-essential heavy metal, presents potential environmental and human risks, is a toxic and carcinogenic metal, and can lead to elevated ROS formation, which in turn leads to DNA damage, as well as its ability to interfere with cell signaling [23].
We noted that the lead concentration in diabetic patients was lower than the healthy persons. This result disagreed with other studies like Akinloye et al. who found that the mean value of Pb was significantly higher in the serum of diabetic patients when compared with the control [21] and Salman et al. found that Cd and Pb were relatively high in patients with type 1 diabetes [24].
Serum level of lead for males in diabetic patients was insignificantly different from healthy subjects and this was true for females in our study. These results were consistent with Tadayon et al., who estimated hair lead levels and found that there was a nonsignificant change between type 2 diabetic patients and the control group [18]. Kolachi et al. studied the level of Pb in biological samples (whole blood, scalp hair, and urine) which was found to be increased significantly in diabetic mothers while in our study, the level of Pb was non significantly lower in diabetic females [25].
Excessive exposure to lead has many negative effects on human health. Evidence indicates that exposure to lead increases susceptibility to cancer and may affect human health by interfering with the DNA repair machinery after genetic insults, impairing the ability of cells to develop appropriate and accurate responses to environmental endotoxins [26]. The behavior of lead in our study was the opposite of some international studies [27] might be due to the small sample size or due to biological factors, human nature, and their relationship to geographical distribution. This can be considered the first study of diabetic patients in Iraq (Al-Najaf Governorate, Al-Hira region) for the elements cadmium and lead so no other local study for results comparison.
The serum concentration of cadmium, in our research, was significantly higher in Healthy Type1 and Type 2 versus the healthy group, this agreed with many international studies as Salman et al. found that Cd was relatively high in patients with Type 1 diabetes [24] and Kolachi et al. who found that Cd in biological samples (whole blood, scalp hair, and urine) of insulin-dependent diabetic mothers were significantly higher as compared to mothers in the control group [25].
Cadmium as a heavy metal had an important role in the pathogenesis of both types of diabetes. The pancreatic tissue of diabetic rats exposed to Cd for 30 days revealed severe degeneration, necrosis, degranulation, shrinkage, and depression in the islets of Langerhans [28].
Li et al. investigate the effects of long-term exposure to Cd in a type-2 diabetes mellitus mouse model and the underlying mechanism, he concluded that Cd exposure disturbed glucose metabolism and exacerbated diabetes [29].
Another explanation is that Cd exposure in type I diabetic rats causes an increment in oxidative stress and Metallothionine gene expression levels [30].
Our research was the first in terms of measuring these two dangerous heavy metals (Cd and Pb) in the serum of diabetic patients who lived in (Al-Najaf Governorate, Al-Hira region) in Iraq, we recommend other studies for measuring them in other governorates and other biological samples as hair and urine for comparison.
Conclusion
The Cd concentration is low in type1 and types 2 diabetic patients of both genders while the concentration of Pb is lower in diabetic patients than in healthy subjects.
Acknowledgments: The researchers would like to extend their thanks and appreciation to all the workers in the Department of Physics, Faculty of Science, University of Kufa, and all faculty staff for their continuous support in completing the research requirements.
Ethical Permissions: The current study has been approved by the ethics committee of the Faculty of Sciences, University of Kufa.
Conflicts of Interests: All authors declare that there is no conflict of interest in the study.
Authors’ Contributions: Mohammad AR (First Author), Introduction Writer/Main Researcher (30%); Hussein HA (Second Author), Assistant Researcher/Statistical Analyst (20%); Kadhim SA (Third Author), Assistant Researcher (20%); Hashoosh MH (Forth Author), Assistant Researcher/Statistical Analyst (10%); Hassan ES (Fifth Author), Assistant Researcher/Discussion Writer (20%)
Funding/Sources: This is a self-funded study. All authors participated in the costs.
Diabetes mellitus (DM) is a group of metabolic disorders characterized by high blood sugar [1]. It is a chronic disease that occurs when the body is not able to use insulin effectively or when the pancreas does not excrete enough insulin [2]. DM is a global disease that spreads all over the world at a rate that differs from country to country [3]. This disease is usually divided into two types, are the most common, type1 (destruction of β cells with an absolute lack of insulin), and the percentage of people with this type is 5-10%, while the second type is the most prevalent 90-95% of cases, the subjects suffer from a relative lack of insulin secretion [4]. Minerals and trace elements are essential micronutrients required for the normal functioning body. These elements are particularly beneficial for physiological functions [5]. A relationship has been reported between trace elements and diabetes mellitus [6]. Minerals and trace elements are essential for many biochemical reactions, and present as stabilizing components of enzymes and proteins in addition they function as cofactors for many enzymes. Certain trace elements regulate crucial biological processes by binding to the receptor site of the cell membrane or by changing the shape of the receptor to prevent the entry of particular molecules into the cell [7]. Some essential trace metals including copper (Cu), zinc (Zn), magnesium (Mg), iron (Fe), and calcium (ca) are associated with diabetes mellitus type 2 [8]. There are many trace elements altered in diabetes mellitus and might have specific roles in the pathogenesis and progress of this disease, they reported higher levels in the blood, urine, and scalp hair of diabetics than nondiabetics [9]. Lead (Pb) is harmful to most of the human body organs and obstructs metabolism and cellular activities [10]. A linear relationship is reported between blood Pb level and renal dysfunction in age-related diseases after recurrent exposures to Pb [11]. It badly affects the antioxidant pathways [12]. The metal-induced toxicity may cause derangement of antioxidant mechanisms in living tissues; as a result, highly reactive oxygen species (ROS) are produced. This antioxidant discrepancy might lead to the degradation of proteins, nucleic acids, and lipid peroxidation. An oxidative attack of cellular components by ROS is concerned with the pathogenesis of numerous human diseases including diabetes [13].
Cadmium (Cd) is a heavy metal that is broadly detected in air, water, and soil. Increased Cd level in water is absorbed by plants, animals, and humans [14]. Frequent exposure to Cd supplied to its excess addition in the kidney causes renal damage and nephropathy [15]. The high level of Cd reduces calcium absorption, which becomes an approaching cause of bone and kidney losses, called Itai-Itai disease. The Cd might down-regulate glucose transporter-4 (GLUT4) translocation by insulin [16].
A previous study proved that blood Cd and Pb levels were higher in diabetics with reduced antioxidant concentration, and it suggested that the elevation of toxic metals may be a contributing factor to the pathogenesis of diabetes mellitus [17].
These studies have been done in some areas of the world, but these studies in Iraq are very few and this is the first study in the Al-Hira area. Therefore, this study aimed to investigate the concentration of these two elements in different serum samples for diabetic patients and compared them with healthy persons.
Materials and Methods
This pilot study was conducted in the Al-Hira area, Najaf governorate, Iraq from June 2021 to January 2022. It included 25 diabetic patients (12 males and 13 females) and 15 healthy non- diabetic subjects (7 males and 8 females) whose ages ranged between 30-80 years. The diabetic patients with an average age of 52.52 years, have type 1 or type 2 diabetes. Healthy people have an average age of 53 years.
The blood samples were taken after fasting one night and centrifuged after acid digestion to obtain serum which was stored for later use. Lead and Cadmium measurements were done by using an atomic absorption spectrophotometer, SHIMADZU model AA-7000 atomic absorption spectrophotometer (AAS) made in Japan which was calibrated to cadmium and lead elements to obtain values controlled for the groups under study as in Diagrams 1 & 2.
Diagram 1) Calibration of the atomic absorption system for cadmium
Diagram 2) Calibration of the atomic absorption system for lead
Microsoft Excel 2019 was used for spreadsheets, SPSS software version 20 was used to analyze the data, and used the independent t-test and Pearson Correlation to find the statistical significance between the different samples which depend on p<0.05.
Findings
The study population included 25 diabetic patients (12 males and 13 females) and 15 healthy non- diabetic subjects (7 males and 8 females) so the gender is comparable in number between case and control groups. Age ranged between 30-80 years. The diabetic patients had an average age of 52.52 years and the healthy people had 53 years’ average age with a non-significant difference between the two groups. All the participants were living in the Al-Hira area, Najaf governorate in Iraq, and this was the first study for these heavy elements in that area.
This study demonstrated that the cadmium concentration in diabetic patients was higher than in the control group with a high statistical significance (p<0.0001), while the lead concentration is significantly lower in the diabetic group as compared to the healthy group (Table 1). According to our results, the cadmium concentration in diabetic male patients was significantly higher (p<0.001) than in the male of the control group, meanwhile lead concentration in the serum of diabetics was insignificantly (p>0.05) lower than in healthy male persons (Table 1; Diagram 4). The same results were found when comparing concentrations of Cd and Pb in the serum of the female diabetic and control group (Table 1).
Table 1) Concentrations of Cd and Pb elements in the serum of healthy and diabetic patients
Pearson correlation analysis was carried out to determine the relationship between variables (Table 2). There was a strong inverse relationship between cadmium and lead in healthy people, with a statistically significant correlation. There was also a strong inverse and statistically significant association between cadmium in healthy subjects with cadmium in people with diabetes, and a moderate positive correlation between cadmium in healthy people and lead in diabetic patients without statistical significance, and finally, the correlation was negative and not statistically significant between lead in healthy people and cadmium in diabetic patients with the negligible correlation between cadmium among sick and healthy subjects.
Table 2) Comparative study for Pearson correlation r between (Cd and Pb)
In Table 3, there was no statistically significant between the different groups of the studied serum samples for the lead trace. There was statistically significant between healthy versus type 1 and healthy versus type 2 Groups using the independent test as well as for the triple groups (control, type 1, type 2) using the ANOVA test (p<0.05).
Table 3) Comparison of serum samples between healthy and types 1 and 2 diabetes groups
Diagram 3) Illustration of Cadmium and lead serum levels in healthy, type1, and type 2 subjects
Discussion
According to the studied samples that we have in our study, cadmium concentration (for both males and females) in diabetic patients is higher than the healthy ones These results were in agreement with other clinical research which found that Cd level is increased in diabetic and prediabetes patients [17-20].
A cohort study in China suggested that high chronic Cd exposure for general population adults might contribute to elevated fasting blood glucose and result in type 2 diabetes mellitus development [21].
Both experimental and epidemiological studies showed that exposure to the environmental pollutant cadmium is associated with hyperglycemia, type 2 diabetes, and reduced serum insulin. Individual cellular effects of cadmium are reviewed, it is likely that no single mechanism is involved, somewhat multiple mechanisms exist, and work synergistically ensuing in islet dysfunction and finally dysglycemia [22].
it was found that Cadmium, a non-essential heavy metal, presents potential environmental and human risks, is a toxic and carcinogenic metal, and can lead to elevated ROS formation, which in turn leads to DNA damage, as well as its ability to interfere with cell signaling [23].
We noted that the lead concentration in diabetic patients was lower than the healthy persons. This result disagreed with other studies like Akinloye et al. who found that the mean value of Pb was significantly higher in the serum of diabetic patients when compared with the control [21] and Salman et al. found that Cd and Pb were relatively high in patients with type 1 diabetes [24].
Serum level of lead for males in diabetic patients was insignificantly different from healthy subjects and this was true for females in our study. These results were consistent with Tadayon et al., who estimated hair lead levels and found that there was a nonsignificant change between type 2 diabetic patients and the control group [18]. Kolachi et al. studied the level of Pb in biological samples (whole blood, scalp hair, and urine) which was found to be increased significantly in diabetic mothers while in our study, the level of Pb was non significantly lower in diabetic females [25].
Excessive exposure to lead has many negative effects on human health. Evidence indicates that exposure to lead increases susceptibility to cancer and may affect human health by interfering with the DNA repair machinery after genetic insults, impairing the ability of cells to develop appropriate and accurate responses to environmental endotoxins [26]. The behavior of lead in our study was the opposite of some international studies [27] might be due to the small sample size or due to biological factors, human nature, and their relationship to geographical distribution. This can be considered the first study of diabetic patients in Iraq (Al-Najaf Governorate, Al-Hira region) for the elements cadmium and lead so no other local study for results comparison.
The serum concentration of cadmium, in our research, was significantly higher in Healthy Type1 and Type 2 versus the healthy group, this agreed with many international studies as Salman et al. found that Cd was relatively high in patients with Type 1 diabetes [24] and Kolachi et al. who found that Cd in biological samples (whole blood, scalp hair, and urine) of insulin-dependent diabetic mothers were significantly higher as compared to mothers in the control group [25].
Cadmium as a heavy metal had an important role in the pathogenesis of both types of diabetes. The pancreatic tissue of diabetic rats exposed to Cd for 30 days revealed severe degeneration, necrosis, degranulation, shrinkage, and depression in the islets of Langerhans [28].
Li et al. investigate the effects of long-term exposure to Cd in a type-2 diabetes mellitus mouse model and the underlying mechanism, he concluded that Cd exposure disturbed glucose metabolism and exacerbated diabetes [29].
Another explanation is that Cd exposure in type I diabetic rats causes an increment in oxidative stress and Metallothionine gene expression levels [30].
Our research was the first in terms of measuring these two dangerous heavy metals (Cd and Pb) in the serum of diabetic patients who lived in (Al-Najaf Governorate, Al-Hira region) in Iraq, we recommend other studies for measuring them in other governorates and other biological samples as hair and urine for comparison.
Conclusion
The Cd concentration is low in type1 and types 2 diabetic patients of both genders while the concentration of Pb is lower in diabetic patients than in healthy subjects.
Acknowledgments: The researchers would like to extend their thanks and appreciation to all the workers in the Department of Physics, Faculty of Science, University of Kufa, and all faculty staff for their continuous support in completing the research requirements.
Ethical Permissions: The current study has been approved by the ethics committee of the Faculty of Sciences, University of Kufa.
Conflicts of Interests: All authors declare that there is no conflict of interest in the study.
Authors’ Contributions: Mohammad AR (First Author), Introduction Writer/Main Researcher (30%); Hussein HA (Second Author), Assistant Researcher/Statistical Analyst (20%); Kadhim SA (Third Author), Assistant Researcher (20%); Hashoosh MH (Forth Author), Assistant Researcher/Statistical Analyst (10%); Hassan ES (Fifth Author), Assistant Researcher/Discussion Writer (20%)
Funding/Sources: This is a self-funded study. All authors participated in the costs.
Keywords: