Aim: The aim was to look into urinary elimination of Fe, Cu and Zn and their relation with serum insulin in diabetes worlds. Materials and Methods: 40 patients with non-insulin-dependent diabetes, 30 patients with insulin-dependent diabetes and 20 normal healthy topics were investigated for their serum degrees of glucose and insulin and urinary elimination of Fe, Cu and Zn. There was important addition in the urinary elimination of all the three hint elements in diabetic patient. No important correlativity could be found between serum insulin and urinary elimination of Fe, Cu and Zn.
Consequences: The consequences indicate that there is increased elimination of hint elements in diabetes mellitus but it has no relation with serum insulin.
Keywords: Diabetes mellitus, Serum insulin, Trace elements ( Fe, Cu and Zn ) .
Introduction: Trace elements are inorganic elements, which are required in the diet in really little sums, less than 100 mg/day1 and are present in our tissues in merely highly little measures, with the concentration of mcgs to pictograms per gm of moisture organ.2For this ground, they are referred to as hint elements.
Trace elements have indispensable function in normal growing, development and wellness of worlds. They have their function in diverse activities such as haem synthesis, connective tissue metamorphosis, bone development and as indispensable constituent of many enzymes.3In add-on hint elements have besides been shown to act upon the hormone system. They influence the hormone system in different ways depending upon whether the metal is present in surplus or in low province. Therefore, increasing or diminishing the concentration of several hint metals has been shown to act upon the hormonal synthesis, secernment, hormonal precursor consumption into mark secretory organs, endocrine binding to aim tissues and its utilization.4
Diabetess mellitus is most common endocrine disease of worlds and is due to absolute or comparative lack of endocrine, insuline5.Diabetes mellitus is said to be of multifactorial etiology and three meshing mechanisms are thought to be responsible: familial exposure, autoimmunity and environmental factors. Although no definite environmental agent has been identified, the environmental factors implicated include viruses, fleshiness, gestation and vague environmental agents6.
Research findings have emphasized that lack of certain hint elements consequences in diabetes like conditions in experimental animate beings. The hint elements, the lack of which has been shown to ensue in diabetes like symptoms are chromium7,8, zinc9, manganes10,11 and copper12. As the lack of these hint minerals resulted in diabetes like symptoms and the supplementation of the peculiar hint component readily reversed the ascertained symptoms, it can be concluded, from such surveies, that lack of these hint elements adversely influences the synthesis, secernment or action of insulin.
Merely as the hint elements have been shown to act upon the insulin, alterations in the concentration of insulin have besides been shown to act upon or to be associated with alterations in the concentrations of several hint metals in the blood, urine and other tissues. Changing degrees of go arounding endocrine may either change the distribution of hint elements in assorted cellular compartments, or may bring forth a displacement of the metals to the tissues which are non usually associated with these concentration of these elements or they may alter renal or hepatic handling of hint elements ensuing in increased urinary or faecal loss of the hint minerals, thereby bring forthing entire organic structure loss4.
Surveies on “ the consequence of diabetes mellitus on hint component metamorphosis ” have largely been carried out in animate beings, experimental theoretical account for insulin-dependent diabetes mellitus.
Such surveies have revealed that experimental diabetes produces alternations in the absorption13, tissue distribution14, metabolism15 and excretion16 of hint elements. Metamorphosis of Zn, Cu and Fe is greatly influenced and the hormonal instability nowadays in insulin-dependent diabetes is said to be involved in altered hint metabolism14,15. Surveies on human existences are really limited and even the survey which have been carried out, has failed to see any relationship between urinary elimination of hint elements and serum insulin. If it is true that the hormonal instability nowadays in insulin-dependent diabetes is involved in changing hint component excretion16, so there may be a relationship between urinary elimination of hint elements and serum insulin.
The present survey was hence, aimed to look into the serum insulin and urinary elimination of hint elements ( Fe, Cu and Zn ) in normal, insulin-dependent and non-insulin-dependent diabetes in an effort to happen any correlativity between serum insulin and urinary elimination of Fe, Cu and Zn. Study besides included the comparing of all these parametric quantities among diabetic and normal topics.
MATERIALS AND METHODS:
70 human diabetics, including both males and females, on out-of-door intervention at assorted outpatient sections and at diabetic clinic, DHQ, Mirpur, AJ & A ; K, Peshawar were studied. Diabetic patients had fasting plasma glucose degree of more than 140 mg/dl or random blood glucose degree of more than 200 mg/dl on more than one juncture.
Patients were divided into two groups depending on manner of intervention: Insulin-dependent and non-insulin-dependent.
20 normal healthy topics with no household history of diabetes were taken as controls.
Both control and diabetic topics fulfilled the undermentioned standards:
Their ages ranged from 25-60 old ages. They were normotensives, with normal nephritic maps and no albuminuria. None of these had open grounds of liver, enteric or endocrinal upset. Subjects holding steroids, or taking any vitamin or mineral supplementation were non included in the survey.
Patients were asked to stop insulin for 24-48 hours and unwritten hypoglycaemic drugs for 5 yearss before roll uping the urine samples.
Collection of blood samples:
Blood was drawn from each patient and normal topic after 10-12 hours fast from an antecubital vena between 8.00-10.00 a.m. Serum was separated within one hr. Glucose appraisal was carried out immediately. Serum for insulin appraisal was stored at -20A°C boulder clay analysis.
Collection of urine samples:
24 hours urine samples were collected in 4-5 litre plastic containers, washed with 15 % HCl and rinsed thrice with deionized H2O. Each patient was provided with a labeled container and a bag in which to transport the container at his or her visit to the infirmary and at the same clip, the patient was asked to stop the intervention, and to get down urine aggregation after 5 yearss in instance of unwritten hypoglycaemic drugs and 24-48 hours, in instance the patient was taking insulin. On the twenty-four hours of blood trying container was collected and the patient was asked about figure of times he or she had missed invalidating into the aggregation container. Merely the urine aggregations reported as complete were used in the analysis. Immediately after aggregation, the volume of each specimen was recorded.
20 milliliter of each urine specimen was centrifuged for 10 proceedingss to take bacteriums, cells, dramatis personaes and other particulate stuffs. 15 milliliter of the supernatant was decanted in plastic bottles and stored at -20A°C boulder clay analysis. It was used for appraisal of Fe, Cu and Zn.
1ml of piss was used for sensing of proteins by chemical method utilizing sulfosalicylic acid. All the glass and plastic ware used were soaked in 15 % HCl overnight and so rinsed thrice with deionized H2O.
Serum glucose was measured by glucose oxidase/peroxidase colorimetric method utilizing analytical kit ( Boehringer Manheim GmbH Diagnostica ) . Serum insulin was measured by RIA utilizing analytical kit provided by Diagnostic Products Corporation, Los Angles, USA.
Urinary Fe was measured by method of Olson and Hamlim17, likewise as for serum. Urinary Cu and Zn were measured by method of Dawson et Al. ( Cited by Varley ) 18. Atomic soaking up spectrophotometer model3030-B Perkin Elmer Company, Norwalk was used for the three hint elements.
Students paired’t ‘ trial was used for statistical analysis.
AGE, SEX AND WEIGHT OF NORMAL SUBJECTS AND DIABETIC PATIENTS:
The tabular array shows the age, sex and organic structure weight of normal and diabetic patients. The values are average A± s. e. m. The entire figure of topics is given in parametric quantities.
The patients enduring from diabetes were grouped harmonizing to the manner of intervention. The average serum values A± s. e. m for glucose and insulin in different groups are given. Number of topics is given in parentheses.
Fast SERUM IRON, COPPER AND ZINC IN NORMAL SUBJECTS AND DIABETIC PATIENTS
Serum Fe, Cu and Zn were measured by atomic soaking up spectrophotometery. Concentrations are expressed as mean values A±s. e. m. Number of topics is given in parentheses.
P & lt ; 0.001 = The Valuess are extremely important as compared to normal control topics.
COMPARISON OF 24-HOUR URINE VOLUME, URINARY IRON, COPPER AND ZINC IN NORMAL SUBJECTS AND DIABETIC PATIENTS
The values are expressed as average A± s. e. m. Number of topics is parentheses.
P & lt ; 0.001 = Valuess are extremely important as compared to command topics.
Table 1 describes the ages, sex and weight of normal topics and diabetic patients. Age and organic structure weight of insulin-dependent diabetics showed no important difference. While in non-insulin-dependent diabetics it was higher ( p & lt ; 0.001 ) as compared to controls.
Fasting serum glucose and immunoreactive insulin in normal topics and diabetic patients are shown in table 2. There is significance addition in serum glucose degrees in diabetic patients ( p & lt ; 0.001 ) and this addition is more pronounced in insulin-dependent diabetics as compared to non-insulin-dependent diabetics.
Serum insulin is significantly increased in non-insulin-dependent diabetics ( P & lt ; 0.001 ) , while in insulin-dependent diabetics, the serum insulin is significantly decreased ( p & lt ; 0.001 ) as compared to normal controls.
Table 3 describes the 24 hours urinary volume and elimination of Fe, Cu and zinc/24 hours in normal topics every bit good as diabetic patients. The consequences of this tabular array indicate that urine volume is significantly increased in diabetic patients as compared to normal topics ( P & lt ; 0.001 ) and this is significantly higher in insulin-dependent diabetics as compared to non-insulin-dependent diabetics ( P & lt ; 0.001 ) . 24 hours urinary elimination of Fe, Cu and Zn is besides significantly increased in both the classs of diabetes ( P & lt ; 0.001 ) , with significantly more elimination in insulin-dependent diabetics ( P & lt ; 0.001 ) than non-insulin-dependent diabetics.
Table 4 gives the urinary elimination of Fe, Cu and Zn per millilitre, in normal and diabetic topics. The consequences indicate that urinary elimination of Fe, Cu and Zn per millilitre is besides significantly increased in diabetics as compared to controls and once more this addition is significantly more pronounced in insulin-dependent diabetics than in non-insulin-dependent diabetics.
Marked alterations in hint component metamorphosis have been demonstrated in streptozotocin-diabetic rate14,15.
Prolonged streptozotocin induced diabetes besides interferes, with normal form of bone mineralization19.
Increased elimination of Fe, Cu and Zn has been reported in experimental diabetes in rates16. Surveies of triumph et al.20 provided direct grounds of influence of hormonal instability on hint metals.
Our findings support and widen the old work. Zinc elimination among normal topics is 420.07A±14.7 Aµg/day and is in conformity with the by and large stated average Zn elimination of 300-600 Aµg/day18. Similarly urinary elimination of Fe and Cu is besides within the normal international mention scope.
Consequences besides demonstrate that diabetic status is associated with increased 24 hours urinary elimination of the three micronutrients zinc, Cu and Fe. Among the diabetics in both insulin-dependent and non-insulin-dependent diabetics average elimination was in surplus of 600 Aµg/day. Similarly Fe and Cu elimination was besides significantly higher in both the insulin-dependent and non-insulin-dependent diabetics as compared to controls. Hyperzincuria has antecedently been reported in diabetic humans184.108.40.206.
Although exact mechanism of increased urinary elimination has non been elucidated, assorted possible mechanisms have been proposed. Diabetic status consequences in increased glomerular filtration rate and it is suggested that such an change in nephritic map may lend to the increased urinary losingss of the hint elements16. But, the observation that addition in urine volume brought approximately by imbibing extra H2O did non change the entire urinary Zn elimination in non-diabetics21 would on the other manus indicate that the polyuria of diabetes may non be an of import determiner of Hyperzincuria or increased elimination of other hint elements in diabetes. This is farther supported by fact that when the concentrations were expressed per millilitre of piss, statistically important differences were still observed.
Increased urinary loss of Fe Cu and Zn could be explained by increased dietetic consumption but urinary loss of Fe, Cu and Zn varies small with dietetic consumption because the predominant healthy excretory path in healthy adult male is the enteric piece of land. Increased urinary elimination reflects acute alterations in biologic stores25. No addition in urinary Zn has been seen when dietetic consumption is doubled in human topics. The surplus is eliminated via the feces24. The possibility of difference between diabetics and normal in their selective soaking up of hint elements in the intestine can non be ignored but information on this facet is non available.
Pidduck et al.24 has proposed three possible mechanisms.
It could be that Hyperzincuria or increased elimination of Fe and Cu indicates an abnormalcy of production or dislocation of metalloenzymes or metal-enzymes composites.
It is possible that some portion of organic structure is bring forthing its ain metabolites with chelating belongingss, it could be the pancreas of the diabetic which is disorganized in the manner, suggested.
Family surveies suggest a important heritability of urinary Zn elimination in 19 households. Some households be givening to be low urinary Zn excreters and others high. It would look possible that urinary elimination of Zn or perchance other hint elements is controlled by allelomorphs at a figure of venue and that diabetics possess a different mixture of allelomorphs to those possessed by non-diabetics.
It is besides believed that increased elimination is chiefly of endogenous beginning for illustration, musculus breakdown16 or diabetic bone loss. However, the bone mineral loss in diabetics can merely account for less than 10 % of the diabetic hyperzincuria22. Urinary Zn losingss are specially increased during katabolic states26.
Intense exercising may increase urinary losingss and it has been shown that exercising additions skeletal musculus protein dislocation and such phenomena could increase urinary Zn losses27. Zinc elimination is shown to increase under assortment of katabolic conditions and Zn is reported to be derived from skeletal musculus. Although the surveies have been carried out for Zn, the skeletal musculus protein breakdown my lead to increased, urinary elimination of Fe and Cu.
Untreated diabetes may stand for a katabolic province and addition in skeletal musculus protein turnover and it is likely that urinary elimination of Fe, Cu and Zn may be derived from skeletal musculus tissue.
As none of the patients had albuminurias, the increased urinary losingss of these trace metals may possibly be due to altered hormone position seen in diabetes and non due to diabetic kidney disease. This decision is supported by fact that improved metabolic control in diabetic rats by insulin intervention resulted in significantly reduced elimination of Fe, Cu and Zn in the urine16 in diabetic worlds in the Restoration of normozincuria16.
No relation could be found between serum insulin and day-to-day urinary elimination of Fe, Cu and Zn in either normal persons or diabetic patients. This may be due to the ground that endocrine relationships in uncontrolled diabetes become extremely complex because glucagon and glucocorticoids degrees are increased in response to insulin lack. Both of these are shown to act upon the serum or urinary degrees of hint elements. Other ground may be that the go arounding immunoreactive insulin in diabetics may non be active biologically.
Non-insulin-dependent diabetes had significantly higher organic structure weight every bit compared to controls. Association between increased organic structure weight and no-insulin-dependent diabetes has antecedently been described26. Obesity is of much greater importance than either race or sex in the etiology of non-insulin-dependent diabetes29. Therefore, a recommendation against going fleshy might be given. A sum of 25 patients ( 35.7 % ) out of 70 had a positive household history of diabetes. High per centum of patients holding positive household history of diabetes suggests a insouciant relationship of this factor with the disease.
This survey may hold clinical and pathological deductions. If there is increased elimination of hint elements in diabetes, this may connote that when dietetic degrees are deficient, the patient ‘s tissues are depleted in favour of increased urinary elimination. In this instance, it is speculated that physiological degrees of these foods may be low and some of the pathological events seen in diabetics may be related to or partially explained by lack of these hint elements. For illustration, there is increased incidence of inborn deformities in diabetic pregnancy30 and lack of Cu and Zn has besides been reported to be associated with inborn malformations31.
Copper lack is associated with impaired collagen synthesis and increased incidence of breaks. Increased incidence of self-generated breaks has besides been reported in diabetes19. But verification of cause and consequence relationship of these phenomena necessesitates farther surveies.