|Year : 2019 | Volume
| Issue : 5 | Page : 1032-1037
|Effects and association of pro-oxidants with magnesium in patients with diabetic nephropathy
Kamal Kachhawa1, Poonam Kachhawa2, Divya Agrawal3, Sanjay Kumar4, Purnima Dey Sarkar5
1 Department of Biochemistry, Government Medical College, Datia, Madhya Pradesh, India
2 Department of Biochemistry, Saraswathi Institute of Medical Sciences, Hapur, Uttar Pradesh, India
3 Department of Anatomy, GSL Medical College, Rajahmundry, Andhra Pradesh, India
4 Department of Pharmacology, GSL Medical College, Rajahmundry, Andhra Pradesh, India
5 Department of Biochemistry, Mahatma Gandhi Memorial Medical College, Indore, Madhya Pradesh, India
Click here for correspondence address and email
|Date of Submission||12-Sep-2018|
|Date of Decision||28-Oct-2018|
|Date of Acceptance||29-Oct-2018|
|Date of Web Publication||4-Nov-2019|
| Abstract|| |
Diabetic nephropathy (DN) is the most common microvascular complication observed in patients with type-2 diabetes mellitus. Furthermore, magnesium (Mg) deficiency is a common problem in diabetic patients. In this study, we estimated the levels of Mg, which is an important trace element and pro-oxidant marker, and then evaluated the association between serum Mg and pro-oxidants in patients with DN. In the present study, 200 patients were enrolled and were divided into two groups. The control and DN groups consisted of 100 healthy individuals and 100 patients with DN, respectively. Serum Mg, total anti-oxidant capacity (TAC), and superoxide dismutase (SOD) levels were estimated using the Calmagite, Koracevic, and Marklund and Marklund methods, respectively. Glutathione (GSH) and malondialdehyde (MDA) levels were estimated using the Tietze F and Jean CD method, respectively. Mg levels were found to be significantly decreased in the DN group in comparison to the control group. Anti-oxidant markers were statistically significantly reduced (P <0.001), whereas MDA levels were statistically significantly elevated (P <0.001) in the DN group compared to the control group. There was a significant positive association of Mg with TAC, SOD, and GSH. A statistically significant negative association of Mg with MDA (r = −0.302, P <0.001, n = 100) was also observed. An apparent relationship was observed between hypomagnesemia and oxidative stress in patients with DN. Lower levels of Mg and oxidative stress were also strongly linked.
|How to cite this article:|
Kachhawa K, Kachhawa P, Agrawal D, Kumar S, Sarkar PD. Effects and association of pro-oxidants with magnesium in patients with diabetic nephropathy. Saudi J Kidney Dis Transpl 2019;30:1032-7
|How to cite this URL:|
Kachhawa K, Kachhawa P, Agrawal D, Kumar S, Sarkar PD. Effects and association of pro-oxidants with magnesium in patients with diabetic nephropathy. Saudi J Kidney Dis Transpl [serial online] 2019 [cited 2019 Nov 21];30:1032-7. Available from: http://www.sjkdt.org/text.asp?2019/30/5/1032/270257
| Introduction|| |
Diabetic nephropathy (DN) is the most common cause of microvascular complications observed in patients with type-2 diabetes mellitus (T2DM). DN is a progressive disease that takes a long time to develop. Primary symptoms of DN include increased excretion of urinary albumin and glomerular hyperfiltration. In this study, we estimated the levels of magnesium (Mg), an important trace element and a pro-oxidant marker, total antioxidant capacity (TAC), superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) and then evaluated their association in patients with DN. Mg is a chief electrolyte of physiological importance; it is one of the most important intracellular cations in the cells. Significant changes in Mg levels can be observed in patients with DN. An increased prevalence of Mg deficiency has been identified in patients with DN, especially in those who have poorly controlled glycemic profiles, with longer duration of the disease, and with the presence of chronic micro-vascular complications. However, the antioxidant defense system of the body reduces damage by decreasing oxidative stress. The most common cause is diabetes or hyperglycemia, which increases the production of reactive oxygen species (ROS), causing cellular dysfunction and damage, ultimately resulting in diabetic microvascular complications.
| Materials and Methods|| |
The study was performed in the Biochemistry Department at the Sri Aurobindo Institute of Medical Sciences (SAIMS) and Hospital. The present study included a total of 200 patients, divided into two groups. The control group consisted of 100 healthy individuals, and the DN group consisted of 100 patients with DN. The patients were diagnosed to have DN in the Department of Nephrology at the SAIMS Hospital. They were selected randomly without any bias for age, sex, occupation, socioeconomic status, and duration of disease. The period of study was from January 2015 to November 2016.
The proposal of the study was presented to the Institutional Ethics Committee after the pilot project was successful. The study was approved by the ethical committee of the SAIMS institute. Informed consent was obtained from all patients. There was no active intervention in the treatment protocol of the patients. The principles of Helsinki declaration were strictly adhered to during the entire course of the study.
All patients with DN diagnosed by the Department of Nephrology at SAIMS Hospital in whom consent was obtained for this study and patients with a positive dipstick test for microalbuminuria were included in the study.
All patients with macroalbuminuria, patients on dialysis, and patients suffering from other medical conditions, such as cardiac disease, were excluded.
Blood samples were taken from the antecubital vein following overnight fasting. The blood sample was collected in plain, fluoride, and EDTA-coated vacutainers. The blood sample was centrifuged for 15 min at 3000 rpm at room temperature. The serum was stored at 4°C for biochemical investigations. Serum Mg was estimated by the Calmagite method. TAC was estimated with the D. Koracevic method. SOD was estimated using the Marklund and Marklund method. GSH content in red blood cells was estimated with the Tietze F method. MDA was estimated with the Jean CD method.
| Statistical Analysis|| |
Statistical analysis was performed using the Statistical Package for Social Sciences version 21.0 (IBM Corp., Armonk, NY, USA). Results were expressed as mean ± standard deviation and were analyzed by unpaired Student’s t-test. Pearson’s correlation was used to evaluate the correlations between the variables of the case groups, and significance was set at P >0.05.,,
| Results|| |
The study included 100 healthy controls and 100 patients with DN, matched for age and sex. Both groups underwent complete history and clinical examination. Demographic data, fasting blood sugar (FBS) level, basal metabolic index (BMI), Mg levels, and stress profile examination of both case and control groups are presented in [Table 1]. BMI and FBS statistically significantly increased (P <0.001) in cases compared to the control group. Mg levels statistically significantly decreased (P <0.001) in cases compared to the control group. Anti-oxidant markers (TAC, SOD, and GSH) statistically significantly decreased (P <0.001), and MDA levels significantly increased in cases compared to those in the control group.
|Table 1: Demographic parameters, trace elements, and anti-oxidant profile in case and control groups (mean ± standard deviation).|
Click here to view
[Table 2] shows the correlation analyses between Mg and the different variables that are depicted in [Figure 1], [Figure 2], [Figure 3], [Figure 4]. [Figure 1], [Figure 2], and [Figure 4] show a significant positive association of Mg with TAC (r = 0.501, P <0.001, n = 100), SOD (r = 0.287, P<0.001, n = 100), and GSH (r = 0.539, P <0.001, n = 100), respectively. [Figure 3] shows a statistically significant negative association of Mg with MDA (r = −0.302, P <0.001, n = 100).
|Table 2: Association of magnesium levels with pro-oxidant parameters in cases.|
Click here to view
|Figure 1: Correlation between serum magnesium and total anti-oxidant capacity.|
Click here to view
|Figure 2: Correlation between serum magnesium and super oxide dismutase.|
Click here to view
| Discussion|| |
In this study, we found a significant lower level of serum Mg in cases than in the control group. Several other studies have also reported similar results in patients with DN.,, Mg is an intracellular cation and affects several metabolic pathways of glucose utilization. However, the precise mechanism of serum Mg deficiency is not completely known. The mechanism of Mg deficiency, which is directly associated with a depletion of Mg, is based on their utilization. One accepted mechanism based on increased urinary excretion of Mg has been hypothesized to be a result of osmotic diuresis, suggesting an association between diabetes with low Mg levels. Some other important factors include diarrhea, vomiting, and poor sodium intake, which can lead to Mg deficiency in DN patients. Hyperglycemia and insulinemia also increase urinary Mg excretion. Urinary Mg excretion and FBS have been found to be inversely related to serum Mg levels. Hypo-magnesemia is currently considered to be an accurate predictor of progression of DN., Furthermore, a previous study has also demonstrated an inverse relation between Mg intake and DM.
Mg deficiency leads to the activation of the renin–angiotensin system, which also induces oxidative stress. Other studies have also shown that hyperglycemia induced free radical formation, inflammation, apoptosis, and cell death, resulting in compromised anti-oxidant function. In this study, we found a significant correlation between serum Mg and antioxidant profile. TAC, SOD, and GSH have been shown to have a significant positive correlation with Mg, consistent with previous studies. Mechanisms contributing to the formation of free radicals in T2DM may include nonenzymatic and auto-oxidative glycosylation, the inflammatory mediators, and a weak anti-oxidant defense system. However, Mg itself has been reported to possess anti-oxidant properties by scavenging free radicals, probably by affecting the rate of spontaneous oxidation and reduction of superoxide anion., Mg is helpful in the maintenance of GSH concentration to protect against damage due to free radicals in the erythrocyte membrane. Thus, GSH deficiency may impair the activity of antioxidant enzymes as well as chain-breaking aqueous and lipid phase antioxidants in the cell wall. A previous study has also established a strong, direct correlation between Mg levels in red blood cells and circulating reduced/oxidized glutathione concentration (r = 0.84, P <0.0001). Some studies have also found a negative correlation between Mg levels and oxidative stress markers (SOD and MDA) in patients that were chronically exposed to stress. However, similar studies in Korean adults reported no such correlation, which was incon-sistent with our results.
MDA is a by-product formed by lipid peroxidation of cellular polyunsaturated fatty acids. MDA formation is thought to play an important role in the development of late diabetic complications., Erythrocytes from Mg-deficient animals are more susceptible to damage from free radicals. In our study, plasma MDA negatively correlated with serum Mg level, which was indicative of the role of Mg in the generation of ROS, thereby causing lipid per-oxidation. A previous study has also shown a negative correlation between Mg and MDA., Furthermore, hypo-magnesemia is currently considered an accurate predictor of progression of DN.
| Conclusion|| |
Our study demonstrates an apparent relationship between hypo-magnesemia and oxidative stress in patients with DN. Lower levels of Mg and oxidative stress were strongly linked. The results of this study may also be helpful in the formulation of effective antioxidant therapies for patients with early development of DN and better management of the disease. Consequently, we propose that a better understanding of Mg metabolism and effort to minimize hypomagnesemia and stress in the management of patients with diabetes mellitus is required. Therefore, further pre-clinical and clinical studies are necessary to clarify the mechanisms involved in the relationship between Mg deficiency, oxidative stress, and associated complications.
Conflict of interest: None declared.
| References|| |
Kachhawa K, Agrawal D, Rath B, Kumar S. Association of lipid abnormalities and oxidative stress with diabetic nephropathy. J Integr Nephrol Androl 2017;4:3-9. [Full text]
Kachhawa K, Varma M, Kachhawa P, Sahu A, Shaikh M, Kumar S. Study of dyslipidemia and cystatin C levels as a predictive marker of chronic kidney disease in type 2 diabetes mellitus patients at a teaching hospital in central India. J Integr Nephrol Androl 2016;3: 24-8. [Full text]
Barbagallo M, Dominguez LJ. Magnesium metabolism in type 2 diabetes mellitus, metabolic syndrome and insulin resistance. Arch Biochem Biophys 2007;458:40-7.
Mather HM, Levin GE. Magnesium status in diabetes. Lancet 1979;1:924.
Del Gobbo LC, Song Y, Poirier P, Dewailly E, Elin RJ, Egeland GM. Low serum magnesium concentrations are associated with a high prevalence of premature ventricular complexes in obese adults with type 2 diabetes. Cardiovasc Diabetol 2012;11:23.
Kachhawa K, Kachhawa P, Varma M, Behera R, Agrawal D, Kumar S. Study of the stability of various biochemical analytes in samples stored at different predefined storage conditions at an accredited laboratory of India. J Lab Physicians 2017;9:11-5.
] [Full text]
Gindler EM, Heth DA. Colorimetrie determination with bound calmagite of magnesium in human blood serum. Clin Chem 1971;17:662.
Koracevic D, Koracevic G, Djordjevic V, Andrejevic S, Cosic V. Method for the measurement of antioxidant activity in human fluids. J Clin Pathol 2001;54:356-61.
Marklund S, Marklund G. Involvement of the superoxide anion radical in the autoxidation of Pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 1974;47:469-74.
Tietze F. Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: Applications to mammalian blood and other tissues. Anal Biochem 1969;27:502-22.
Dousset JC, Trouilh M, Foglietti MJ. Plasma malonaldehyde levels during myocardial infarction. Clin Chim Acta 1983;129:319-22.
Ankush RD, Suryakar AN, Ankush NR. Hypomagnesaemia in type-2 diabetes mellitus patients: A study on the status of oxidative and nitrosative stress. Indian J Clin Biochem 2009;24:184-9.
Resnick LM, Altura BT, Gupta RK, Laragh JH, Alderman MH, Altura BM. Intracellular and extracellular magnesium depletion in type 2 (non-insulin-dependent) diabetes mellitus. Diabetologia 1993;36:767-70.
Barbagallo M, Gupta RK, Dominguez LJ, Resnick LM. Cellular ionic alterations with age: Relation to hypertension and diabetes. J Am Geriatr Soc 2000;48:1111-6.
Arpaci D, Tocoglu AG, Ergenc H, Korkmaz S, Ucar A, Tamer A. Associations of serum magnesium levels with diabetes mellitus and diabetic complications. Hippokratia 2015;19: 153-7.
Hans CP, Sialy R, Bansal DD. Hypomagnesemia in diabetic patients: Correlation with oxidative stress. Int J Diabetes Dev Ctries 2002;22:122-31.
McNair P, Christensen MS, Christiansen C, Madsbad S, Transbøl I. Renal hypomagnesaemia in human diabetes mellitus: Its relation to glucose homeostasis. Eur J Clin Invest 1982;12:81-5.
Sakaguchi Y, Shoji T, Hayashi T, et al. Hypomagnesemia in type 2 diabetic nephropathy: A novel predictor of end-stage renal disease. Diabetes Care 2012;35:1591-7.
Tin A, Grams ME, Maruthur NM, et al. Results from the atherosclerosis risk in communities study suggest that low serum magnesium is associated with incident kidney disease. Kidney Int 2015;87:820-7.
Kim DJ, Xun P, Liu K, et al. Magnesium intake in relation to systemic inflammation, insulin resistance, and the incidence of diabetes. Diabetes Care 2010;33:2604-10.
Jones DP. Radical-free biology of oxidative stress. Am J Physiol Cell Physiol 2008;295: C849-68.
Kachhawa K, Varma M, Kachhawa P, Agrawal D, Shaikh MK, Kumar S. Study of dyslipidemia and antioxidant status in chronic kidney disease patients at a hospital in South East Asia. J Health Res Rev 2016;3:28. [Full text]
Kuzniar A, Mitura P, Kurys P, Szymonik-Lesiuk S, Florianczyk B, Stryjecka-Zimmer M. The influence of hypomagnesemia on erythrocyte antioxidant enzyme defence system in mice. Biometals 2003;16:349-57.
Tamrakar S, Kachhawa K, Agrawal D, Varma M, Swain TR, Kumar S. Study of trace elements (Mg and Cu) and dyslipidemia in type 2 diabetes mellitus (T2DM) patients presenting in a tertiary care hospital of South East Asia. Int J Curr Res 2016;8:26972-5.
Barbagallo M, Dominguez LJ, Tagliamonte MR, Resnick LM, Paolisso G. Effects of glutathione on red blood cell intracellular magnesium: Relation to glucose metabolism. Hypertension 1999;34:76-82.
Cernak I, Savic V, Kotur J, et al. Alterations in magnesium and oxidative status during chronic emotional stress. Magnes Res 2000;13:29-36.
Bae YJ, Choi MK. Magnesium intake and its relevance with antioxidant capacity in Korean adults. Biol Trace Elem Res 2011;143:213-25.
Zheltova AA, Kharitonova MV, Iezhitsa IN, Spasov AA. Magnesium deficiency and oxidative stress: An update. Biomedicine (Taipei) 2016;6:20.
Kamath S, Varashree BS, John P, Mohapatra N, Shenoy RP, Shetty RK. Correlation of serum magnesium and malondialdehyde levels in patients with myocardial infarction. Res J Pharm Biol Chem Sci 2016;7:791-5.
Varma M, Kachhawa K, Sahu A, Kachhawa P. Association of antioxidant enzymes and MDA level in diabetic nephropathy patients in Indore region of Madhya Pradesh. J Pure Appi Microbiol 2014;8:4137-41.
Department of Pharmacology, GSL Medical College, Rajahmundry, Andhra Pradesh
[Figure 1], [Figure 2], [Figure 3], [Figure 4]
[Table 1], [Table 2]
| Article Access Statistics|
| Viewed||460 |
| Printed||6 |
| Emailed||0 |
| PDF Downloaded||45 |
| Comments ||[Add] |