Intracellular Magnesium Concentration Research Articles

Intralymphocyte free magnesium and calcium and insulin tolerance test in a group of essential hypertensive patients

In order to assess the links which are claimed to exist between peripheral insulin resistance and intracellular magnesium and calcium concentrations, we measured free intralymphocyte magnesium (Mg i) and calcium (Ca i) concentrations as well as the rate constant of plasma glucose disappearance (K itt) after insulin injection (insulin tolerance test: ITT) in a group of 16 normotensive control subjects (NC) and 34 essential hypertensive subjects (EH). Mg i and Ca i were measured in triplicate by means of a fluorimetric technique based on the dyes furaptra and fura-2 respectively. K itt values proved significantly reduced in EH as compared to NC (M ± SD, EH: 4.49 ± 1.31 vs 5.28 ± 1.19, P < 0.05; 95% confidence limits: 0.23–1.5). Mg i and Ca i were not statistically different in EH as compared to NC subjects (Mg i, NC: 266 ± 20 μmol l ; EH: 245 ± 50 μmol l ; Ca i, NC: 47 ± 9 nmol l , EH: 46 ± 13 nmol l ). We found a statistically significant inverse correlation in the whole study group between K itt and body mass index (R = −0.363, P < 0.01) and a statistically significant positive correlation between K itt and Mg i (R = 0.347, P = 0.013) was found. In a step-up multivariate regression analysis including blood pressure, plasma lipids, BMI, plasma magnesium, fasting insulin, fasting glucose, Mg i and Ca i, the dependent variable K itt is statistically significantly correlated with body mass index and Mg i. In a first attempt to study the relationships between insulin resistance, Mg i and Ca i in nucleated cells, the chosen index of peripheral resistance seems to be linked to intracellular free magnesium.

Intracellular divalent cation homeostasis and developmental competence in the hamster preimplantation embryo.

The intracellular magnesium and calcium ion concentrations of in vivo-developed 2-cell hamster embryos were measured using ratiometric fluorometry. Intracellular magnesium and calcium ion concentrations were found to be 0.369 +/- 0.011 mM and 129.3 +/- 7.5 nM respectively. Culture of 1-cell hamster embryos for 24 hr to the 2-cell stage in control medium containing 0.5 mM magnesium and 2.0 mM calcium resulted in approximately a threefold increase to 343.5 +/- 8.0 nM in intracellular calcium ion concentration, while magnesium ion levels were not altered (0.355 +/- 0.007 mM). Increasing medium magnesium concentrations to 2.0 mM significantly increased intracellular magnesium ion concentrations of cultured 2-cell embryos with a concomitant reduction in intracellular calcium ion concentrations. Furthermore, increasing the medium magnesium concentration to 2.0 mM significantly increased development of 1-cell embryos collected at either 3 or 9 hr post-egg activation to the morula/blastocyst and blastocyst stages. Resultant blastocysts had an increased total cell number and increased development of the inner cell mass. Most important, however, culture with 2.0 mM magnesium increased the fetal potential of cultured 1-cells twofold. Therefore, because highest rates of development were observed in a medium that resulted in reduced intracellular calcium ion concentrations, it appears that altered calcium homeostasis is associated with impaired developmental competence of 1-cell embryos in culture.

Bronchial reactivity and intracellular magnesium: a possible mechanism for the bronchodilating effects of magnesium in asthma

1. Increased bronchial smooth muscle contractility with consequent bronchial hyperreactivity are characteristic physiopathological events of asthma. Since magnesium intervenes in calcium transport mechanisms and intracellular phosphorylation reactions, it constitutes an important determinant of the contraction/relaxation state of bronchial smooth muscle. In the present study we investigated the relationship between bronchial reactivity, assessed by methacholine-provocation test, and magnesium concentrations both at extracellular and intracellular levels measured by spectrophotometry. Twenty-two patients with mild-to-moderate asthma and 38 non-asthmatic subjects with allergic rhinitis (24 allergic to Parietaria pollen and 14 allergic to Grass pollen) were recruited to the study. Exclusion criteria included renal failure, hepatic diseases, heart failure and arterial hypertension. 2. The salient finding of our study is that there is a strong positive correlation between bronchial reactivity and the level of intracellular magnesium (r=0.72, P<0.0001), both when the groups are analysed separately or together. Intracellular magnesium concentrations in the group of patients with asthma were significantly lower (1.8+/-0. 01 mmol/l; n=22) when compared with levels in rhinitis subjects allergic to Parietaria (1.9+/-0.01 mmol/l; n=24, P<0.05), and with levels in rhinitis subjects allergic to Grass pollen (2.0+/-0.03 mmol/l; n=14, P<0.05). Serum levels of the ion were similar in all groups. 3. We conclude that the level of intracellular magnesium may be an important determinant of bronchial hyperreactivity, as supported by the significant positive correlation between these two parameters in allergic patients with known bronchial hyperresponsiveness. This finding, in addition to reports of the bronchodilating effects of magnesium administration in patients with asthma, confirms the proposed role of this ion in the pathogenesis and treatment of asthma.

Measurements of Intracellular Mg 2+ Concentration in Mouse Skeletal Muscle Fibers with the Fluorescent Indicator Mag-Indo-1

Measurements of intracellular free magnesium concentration ([Mg 2+] i) were performed on enzymatically isolated skeletal muscle fibers from mice, using the fluorescent ratiometric indicator mag-indo-1. An original procedure was developed to calibrate the dye response within the fibers: fibers were first permeabilized with saponin in the presence of a given extracellular magnesium concentration and were then embedded in silicone grease. The dye was then pressure microinjected into the saponin-permeabilized silicone-embedded fibers, and fluorescence was measured. The results show that for all tested [Mg 2+], the value of the measured fluorescence ratio was higher than that found in aqueous solutions. Furthermore, the apparent binding curve that could be fit to the in vivo ratio data was shifted toward higher [Mg 2+] by a factor of ∼2. Using the in vivo calibration parameters, the mean resting [Mg 2+] i was found to be 1.53 ± 0.16 mM ( n = 7). In an attempt to gain insight into the myoplasmic magnesium buffering capacity, we measured, together with mag-indo-1 fluorescence, the current elicited by the application of carbamylcholine (CCh) to the endplate of isolated fibers, in the presence of a high extracellular magnesium concentration. The results show that, under these conditions, a change in [Mg 2+] i displaying a time course and amplitude qualitatively consistent with the CCh-induced inward current can be measured.

Depolarization triggers intracellular magnesium surge in cultured dorsal root ganglion neurons

Intracellular magnesium concentration ([Mg 2+] i) of cultured dorsal root ganglion (DRG) neurons was measured using the magnesium indicator Mag-Fura-2/AM. [Mg 2+] i was 0.48±0.08 mM (mean±SEM, n=23) at rest, and it increased 3-fold by depolarization with a 60-mM K + solution. The [Mg 2+] i increase was observed in the absence of extracellular Mg 2+, but the increase disappeared in the absence of extracellular Ca 2+. 50 μM cadmium or 100 μM verapamil, a Ca 2+ channel blocker, also diminished the rise of [Mg 2+] i. The additional measurement of an intracellular Ca 2+ concentration ([Ca 2+] i) indicated that the [Mg 2+] i rise requires a threshold concentration of [Ca 2+] i to be reached; above 60 nM. The present results indicate that depolarization induces a Ca 2+-influx through voltage dependent Ca channels and this causes the release of Mg 2+ from intracellular stores into the cytoplasm.

Relationship between blood magnesium and psychomotor retardation in drug-free patients with major depression

SummaryIn previous reports, we have observed that blood magnesium was significantly higher in drug-free patients with major depression when compared to healthy controls. This was especially true for erythrocyte magnesium. Furthermore, the most severely depressed patients had the highest intracellular magnesium content, showing that intracellular magnesium rate was related to the intensity of symptoms. We report here the results of blood magnesium measured in 88 major depressed patients as compared to 61 controls. We show that the mean erythrocyte and also plasma magnesium contents are both increased in these patients. We observe that about 40% of male and female patients have a very significant increase (25%) in intracellular magnesium content as compared to controls. However, about 60% of the hospitalised depressed patients have normal values. None of the controls has high erythrocyte magnesium. This is less evident concerning the plasma magnesium. No differences are observed between patients when classified according to the intensity of moral pain or anxiety. In contrast, the patients with mild to high psychomotor retardation score, which is an index of hypoexcitability, have significant higher erythrocyte magnesium values compared with other patients. The results of male patients without psychomotor retardation do not differ from control values. Our study suggests that central hypoexcitability might be related to an increase in intracellular magnesium observed at the peripheral level, keeping in mind that hyperexcitability, as observed in various conditions such as stress and cardiovascular disorders, is frequently associated, in contrast, with a decrease in blood magnesium.

Cardiac arrhythmias after cisplatin infusion: Three case reports and a review of the literature

Several antineoplastic drugs are known to have cardiactoxic effects [1-4] which may be dose-limiting, and whichrange over a wide spectrum of cardiac pathology, in-cluding cardiomyopathy, ischaemia and arrhythmia [1].Supraventricular and ventricular arrhythmias often oc-cur suddenly and unexpectedly, and are sometimes life-threatening. Their pathogenesis is often ascribed todirect drug-induced myocardial toxicity, but indirectmechanisms due to electrolyte imbalances may also beinvolved [1, 2, 4].Cisplatin is widely used in the treatment of many neo-plastic diseases. Its major side effects include nephrotox-icity, myelotoxicity, ototoxicity, neurotoxicity and gastro-intestinal toxicity [5], but changes in intracellular andextracellular magnesium (Mg) concentrations are alsofrequently reported [6-10]. Conversely, cardiac toxicityis only occasionally observed [11-13], and is commonlyconsidered to be a consequence of direct cisplatin-induced myocardial toxicity [11]. In this paper, we reportthree cases of supraventricular tachyarrhythmia, in whichcisplatin-induced changes in Mg homeostasis may havebeen implicated.CaselIn September 1995, a 68-year-old man was admitted toour department for treatment of a stage HI B non-small-cell lung cancer (NSCLC), without evidence of pericar-dial involvement on computed tomography (CT). Hehad no history of cardiac disease, and his arterial bloodpressure, and basal electrocardiogram (ECG) and echo-cardiography results were normal. Chemotherapy wasstarted with intravenous (i.v.) cisplatin 100 mg/m

Free magnesium concentration in isolated rabbit hearts subjected to high dose isoproterenol infusion: a 31P NMR study

The hypothesis of magnesium deficiency in isoproterenol (ISO) induced myocardial injury has been investigated by 31P nuclear magnetic resonance spectroscopy. High energy phosphate concentrations, pHi, and intracellular free magnesium concentration ([Mg2+]i) were measured in isolated rabbit hearts perfused at constant flow and subjected to 10(-6)M isoproterenol during 30 min. Recent calibrations were used for [Mg2+]i measurements, and uncertainties on [Mg2+]i estimated values were calculated. During isoproterenol infusion, pHi, [PCr], and [ATP] decreased, while [P(i)] increased. When it was stopped, [PCr] completely repleted, whereas only a partial restoration was observed for pHi and [P(i)]. A rise of end-diastolic pressure and perfusion pressure expressed a contracture, concomitant with a lack of [ATP] recovery, which remained at 59 +/- 13% of the rest value. These results establish that 10(-6) M isoproterenol caused severe myocardial injury. [Mg2+]i increased from 0.70 mM at rest to 0.88 mM at the end of the isoproterenol period. Considering the estimated uncertainties on the [Mg2+]i values, this increase was not significant. After isoproterenol infusion, [Mg2+]i progressively decreased to reach 0.72 mM at 45 min recovery. It is concluded that isoproterenol myocardial toxicity may not be related to [Mg2+]i deficiency.

Serum and intracellular magnesium concentrations in intoxicated chronic alcoholic and control subjects

Serum magnesium levels may not accurately reflect overall magnesium homeostasis. To clarify the relationship between serum and intracellular magnesium concentrations in chronic alcoholism, we determined intralymphocytic magnesium levels and serum magnesium, sodium and potassium concentrations in ten chronic alcoholic subjects admitted to the emergency room during acute ethanol intoxication, and compared the results to those of 14 healthy nonalcoholic controls. Serum magnesium, sodium and potassium concentrations were within the normal range in both groups of subjects and determination of intralymphocytic magnesium levels revealed a nonsignificant decrease in alcoholic subjects compared to controls. In conclusion, serum and intralymphocytic magnesium concentrations did not differ between chronic alcoholics and controls in our population; the results of the present study do not support the practice of routine magnesium administration to chronic alcoholics in the emergency room setting.

Magnesium sulphate improves neurologic outcome following severe closed head injury in rats

While recent evidence suggests that brain intracellular free magnesium concentration declines following severe diffuse traumatic brain injury, no studies have examined whether magnesium administration following such injury can improve subsequent neurologic outcome. The present study shows that MgSO 4 administered as a bolus at 30 min following severe closed head injury in rats significantly improves posttraumatic neurologic outcome as assessed by both rotarod and angleboard tests. Moreover, this improvement in outcome was evident with both intravenous and intramuscular drug administration. We conclude that parenteral administration of magnesium sulphate may be neuroprotective following severe closed head injury of a diffuse nature.

Phosphorus J-coupling constants of ATP in human brain.

Proton decoupled 31P NMR spectroscopy of the occipital brain of healthy volunteers was performed with a 1.5 T whole-body imager. By use of two-dimensional chemical-shift imaging in combination with slice-selective excitation well resolved localized spectra (38 ml) were obtained within 34 min from which the homonuclear 31P-31P J-coupling constants of ATP could be determined: J(gammabeta) = 16.1 Hz +/- 0.2 Hz and J(alphabeta) = 16.3 Hz +/- 0.1 Hz (mean +/- SEM, n = 14). Both, the J-coupling constants and the chemical-shift difference between alpha- and beta-ATP (delta(alphabeta) = 8.61 ppm +/- 0.01 ppm) were used to calculate the concentration of intracellular free magnesium. The concentrations are 0.39 mM +/- 0.09 mM by using the average of both coupling constants of each spectrum, which is in fair agreement with 0.32 mM +/- 0.01 mM obtained from the chemical shift of alpha and beta phosphate resonances, which is the more accurate result.

Hypertension, diabetes mellitus, and insulin resistance: the role of intracellular magnesium.

Magnesium is one of the most abundant ions present in living cells and its plasma concentration is remarkably constant in healthy subjects. Plasma and intracellular magnesium concentrations are tightly regulated by several factors. Among them, insulin seems to be one of the most important. In fact, in vitro and in vivo studies have demonstrated that insulin may modulate the shift of magnesium from extracellular to intracellular space. Intracellular magnesium concentration has also been shown to be effective on modulating insulin action (mainly oxidative glucose metabolism), offset calcium-related excitation-contraction coupling, and decrease smooth cell responsiveness to depolarizing stimuli, by stimulating Ca2+-dependent K+ channels. A poor intracellular magnesium concentration, as found in non-insulin-dependent diabetes mellitus (NIDDM) and in hypertensive (HP) patients, may result in a defective tyrosine-kinase activity at the insulin receptor level and exaggerated intracellular calcium concentration. Both events are responsible for the impairment in insulin action and a worsening of insulin resistance in non-insulin-dependent diabetic and hypertensive patients. By contrast, in NIDDM patients daily magnesium administration, restoring a more appropriate intracellular magnesium concentration, contributes to improve insulin-mediated glucose uptake. Similarly, in HP patients magnesium administration may be useful in decreasing arterial blood pressure and improving insulin-mediated glucose uptake. The benefits deriving from daily magnesium supplementation in NIDDM and HP patients are further supported by epidemiological studies showing that high daily magnesium intake to be predictive of a lower incidence of NIDDM and HP. In conclusion, a growing body of studies suggest that intracellular magnesium may play a key role on modulating insulin-mediated glucose uptake and vascular tone. We further suggest that a reduced intracellular magnesium concentration might be the missing link helping to explain the epidemiological association between NIDDM and hypertension.

In vivo study of halothane hepatotoxicity in the rat using magnetic resonance imaging and 31P spectroscopy.

Using magnetic resonance imaging (MRI) and spectroscopy (MRS), in vivo halothane hepatotoxicity was assessed in male Wistar rats. With 1.5% halothane in 100 or 20% O 2, an edematous region, characterized by increased intensity on T 2 weighted images and an increase in regional tissue water content ( ρ water), was seen proximal to the hepatic portal vein in the liver. Both spin–lattice relaxation (T 1) and spin–spin relaxation (T 2) increased in this region, relative to distal regions of the liver. Similarly, a high signal intensity on proton density weighted images was observed in this area. As halothane anaesthesia progressed, a decrease in the adenosine triphosphate–inorganic phosphate ratio (ATP/P i) and an increase in the phosphomonoester–phosphodiester (PME/PDE) ratio was detected in the liver. In addition, intracellular pH decreased and intracellular free magnesium concentration [Mg 2+] increased with time of exposure. Excessive vacuolation, ribosomal disappearance from rough endoplasmic reticulum, mitochondrial swelling and fragmentation of smooth endoplasmic reticulum were observed by transmission electron microscopy (TEM) in samples from the edematous region of the liver.

Intracelluiar Free Magnesium Concentrations in Skeletal Muscle in Chronic Uraemia

Low intracellular free magnesium concentrations ([Mg2+]i) are associated with essential hypertension and may reflect a disordered cellular ionic environment. 31P magnetic resonance spectroscopy was used to study skeletal muscle [Mg2+]i in a group of chronic renal failure (CRF) patients and data were compared with a group of control subjects of similar age. Other data including the patients' blood pressure, medication and plasma biochemistry were collected. There was a significant inverse correlation of [Mg2+]i with systolic (p < 0.001) and diastolic blood pressure (p < 0.05) in the CRF population. In CRF [Mg2+]i was similar (0.52 +/- 0.01 mM, SEM) to controls (0.53 +/- 0.01 mM; p = 0.20), even if just the normotensive patients and controls were compared. There was no correlation of [Mg2+]i with plasma parathyroid hormone, total [Mg2+] or [Ca2+]. Similar to studies in subjects with essential hypertension, these data support a role for [Mg2+]i specifically, and an abnormal intracellular environment more generally, in the pathophysiology of hypertension in CRF.

In vivo 31P NMR spectroscopy of human musculoskeletal tumors as a measure of response to chemotherapy.

The value of in vivo 31P NMR spectroscopy to provide indicators of response to cytostatic chemotherapy was studied in patients with malignant musculoskeletal tumors. Characteristics of untreated cancers were strong signals of PME and PDE, moderately increased Pi and low PCr. The intracellular pH was slightly alkaline. The intracellular concentration of free magnesium was 70% of that in muscle. Spectroscopic findings at different times of therapy were compared with the percentage of tumor necrosis after surgical resection in 28 patients. In follow-up studies, energy-rich phosphates declined in nonresponders, while PME, Pi and frequently PDE increased. Treatment response appeared to involve the reversal of these trends. In five responders, a biphasic pattern was observed, i.e. initially the spectrum changed into that of severely ischemic cell injury followed by a successive phase of apparent 'tumor activation'. Pretreatment levels of (PCr+Pi)/total phosphate > or = 0.35 and PCr/ alpha-NTP > or = 1.5, an accelerated increase in total low-energy phosphates/total high-energy phosphates (> or = 3.0%/day) after the initial drug application, and a long-term decrease (< or = -0.4%/day) during later therapy were highly indicative of tumor response to chemotherapy. Such spectroscopic predictors for treatment response proved to be superior to currently used indices such as tumor size.

Calculation of intracellular cerebral [Mg2+] during hypoxic ischemia by in vivo 31P NMR.

Several algorithms for the calculation of ionized intracellular magnesium concentration from the chemical shifts of MgATP were compared, using in vivo 31P NMR data obtained from swine brain during and following hypoxic ischemia plus i.v. MgSO4 infusion. This analysis reveals that both the absolute ionized intracellular magnesium and relative changes in magnesium may vary widely between algorithms used. The calculated intracellular pH, used in algorithms to determine ionized magnesium concentration was found to be a critical parameter that governs the extent of these differences.

The Regulatory Role for Magnesium in Glycolytic Flux of the Human Erythrocyte

31P NMR was used to measure the intracellular free magnesium concentration ([Mg2+]i) in human erythrocytes while [Mg2+]i was changed between 0.01 and 1.2 mM using the divalent cationophore A23187. 13C NMR and [2-13C]glucose were used to determine the kinetic effects of [Mg2+]i by measuring the flux through several parts of the glucose pathway. Glucose utilization was strongly dependent on [Mg2+]i, with half-maximal flux occurring at 0.03 mM. The rate-limiting step was most likely at phosphofructokinase, which has a Km(Mg2+) of 0.025 mM in the purified enzyme. Phosphorylated glycolytic intermediate concentration was also strongly dependent on [Mg2+]i and [MgATP], and glucose transport plus hexokinase may have been partially rate-determining at [Mg2+]i below approximately 0.1 mM. The pentose phosphate shunt activity was too low to determine the dependence on [Mg2+]i. Phosphoglycerate kinase and 2, 3-diphosphoglycerate mutase fluxes were also measured, but were not rate-limiting for glycolysis and showed no Mg2+ dependence. Human erythrocyte [Mg2+]i varies between 0.2 mM (oxygenated) and 0.6 mM (deoxygenated), well above the measured [Mg2+]i(1/2). It is unlikely, then, that [Mg2+]i plays a regulatory role in normal erythrocyte glycolysis.

Traumatic brain axonal injury produces sustained decline in intracellular free magnesium concentration

Decline in brain intracellular free magnesium concentration following experimental traumatic brain injury has been widely reported in a number of studies. However, to date, these studies have been confined to focal models of brain injury and temporally limited to the immediate 8-h period post-trauma. Recently, a new model of impact-acceleration brain injury has been developed which produces nonfocal diffuse axonal injury more typical of severe clinical trauma. The present study has used phosphorus magnetic resonance spectroscopy and the rotarod motor test to characterise magnesium homeostasis and neurologic outcome over a period of 8 days after induction of severe impact-acceleration injury in rats. Severe impact-acceleration induced injury resulted in a highly significant and sustained decline in intracellular free magnesium concentration that was apparent for 4 days post-trauma with recovery to preinjury levels by day six. There were no significant changes in pH or ATP concentration at any time point post-injury. All animals demonstrated a significant neurologic deficit over the assessment period. The extended period of magnesium decline after severe diffuse brain trauma suggests that repeated administration may be required for pharmacotherapies targeted at restoring magnesium homeostasis.

Relation Between Severity of Magnesium Deficiency and Frequency of Anginal Attacks in Men With Variant Angina

Objectives. We evaluated whether the severity of magnesium deficiency was correlated with the frequency of attacks of variant angina.Background. Magnesium deficiency may be associated with the development of variant angina. However, the relation between the activity of variant angina and magnesium deficiency remains to be elucidated.Methods. We assessed the body magnesium status of 18 men with variant angina: Group 1 (≥4 attacks/week, n = 7) and Group 2 (<4 attacks/week, n = 11). Concentrations of magnesium were determined in serum, urine, mononuclear cells and erythrocytes, and the 24-h magnesium retention rate was determined.Results. Group 1 showed a higher 24-h magnesium retention rate (mean ± SEM 63.5 ± 7.6% vs. 24.9 ± 2.7%, p < 0.01) and a lower intracellular concentration of magnesium in mononuclear cells and erythrocytes than did Group 2 (respectively, 156.3 ± 13.5 vs. 212.1 ± 6.9 fg/cell, p < 0.01; and 3.5 ± 0.5 vs. 5.2 ± 0.4 fg/cell, p < 0.05), demonstrating the presence of magnesium deficiency in Group 1. The 24-h magnesium retention rate and intracellular concentrations of magnesium in mononuclear cells and erythrocytes correlated well with the frequency of anginal attacks (r = 0.78, p < 0.01; r = −0.78, p < 0.01; r = −0.62, p < 0.01, respectively) for all patients.Conclusions. Data suggest that the magnesium status of men with variant angina is closely related to disease activity.

Brain intracellular free magnesium declines after impact-acceleration induced brain injury in rats

Significant declines in brain intracellular free magnesium concentration (Mgf) have been reported following focal traumatic brain injury but not following diffuse axonal brain injury. Since diffuse axonal brain injury is a common feature of clinical trauma, we have used 31P magnetic resonance spectroscopy to determine whether Mgf decline is also a feature of a nonfocal traumatic brain injury model that primarily induces widespread axonal injury. Impact acceleration-induced injury resulted in a highly significant decline in Mgf with an associated decline in cytosolic phosphorylation ratio. There were no significant changes in either ATP concentration or brain pH. We propose that decline in brain Mgf may be a common feature of both focal and diffuse forms of traumatic brain injury and therefore a possible target for therapeutic intervention.