Whether La3+ can enter human peripheral blood lymphocytes by the Na+/Ca2+ exchanger or not and the effect of La3+ on the Na+/Ca2+ exchanger activity are examined by fura-2 technique. And that whether La3+ is sequester...Whether La3+ can enter human peripheral blood lymphocytes by the Na+/Ca2+ exchanger or not and the effect of La3+ on the Na+/Ca2+ exchanger activity are examined by fura-2 technique. And that whether La3+ is sequestered by intracellular organelles (mainly endoplasmic reticulum and mitochondria) is studied by this method. La3+uptake is obviously stimulated by pretreating the cells with ouabain and by removing extracellular Na+, and intracellular La3+concentration ([La3+]i) is directly proportional to its extracellular concentration ([La3+]o). But when [La3+]o exceeds 0.4 mmol/L, the 340/380 nm ratio of fluorescence is no longer varied and the maximum [La3+]i is 1.5X10-12 mol · L-1. The higher concentration of La3+ (0.1 mmol/L) increases Na+/Ca2+ exchange-mediated calcium influx, but lower concentration (10 μmol/L) appears to block calcium influx. The results also suggest that cytosolic La3+ is transported by the ATP-dependent Ca2+ pump. Intracellular Ca2+ stores are depleted by ionomycin, and then ionomycin is added again during the period of La3+ uptake, the 340/380 nm ratio of fluorescence is also increased, these results indicate that La3+ is sequestered by intracellular organelles. A characterization of fura-2-La3+ interaction in solution simulating intracellular ionic composition (pH 7.05) shows that La3+forms a 1: 1 fura-2-La3+complex, and the apparent dissociation constant of La3+ for fura-2 (Kd) is 1.7*10-12 mol · L-1. In addition, the limit of detection of fura-2 for La3+ and Ca2+ is 10-12 and 10-8 mol · L-1 respectively.展开更多
Previously we have shown that 4 Hz and 8 Hz EMF exposures have depressing effect on the thermodynamic activity of water, which decreases peroxide formation. It has also been shown that 4 Hz EMF-treated physiological s...Previously we have shown that 4 Hz and 8 Hz EMF exposures have depressing effect on the thermodynamic activity of water, which decreases peroxide formation. It has also been shown that 4 Hz EMF-treated physiological solution modulates the growth and development of microbes and heart muscle contractility, but 8 Hz EMF has pronounced inhibitory effect on bacterial growth and development. Therefore, in order to elucidate the possible mechanism of 4 Hz and 8 Hz EMF effects on heart muscle function, in the present work the effects of 4 Hz and 8 Hz EMF exposures on heart muscle tissue hydration, the sensitivity of 4 Hz and 8 Hz EMF-induced tissue hydration to 10−4 M ouabain (Na+/K+ pump inhibition) and 10−9 M ouabain (activation of intracellular signaling system) as well as the effects of 4 Hz and 8 Hz EMF exposures on the number of Na+/K+ pump units in the membrane of both young and old rats have been studied. The obtained data allow us to suggest that 8 Hz EMF exposure has more pronounced age-dependent modulation effect on tissue hydration of heart muscle than 4 Hz EMF and this effect is sensitive to Na+/K+ pump activity and intracellular signaling system.展开更多
Results from a systematic experiment on isolated perfused rat heart and isolated myc-cytes of adult rat showed that the mechanism of calcium influx during myocardial ischemia-reperfusion is due to the development of i...Results from a systematic experiment on isolated perfused rat heart and isolated myc-cytes of adult rat showed that the mechanism of calcium influx during myocardial ischemia-reperfusion is due to the development of intracellular sodium overload during ischemic pe-riod, on reperfusion, the high intracellular Na^+ content activated the reverse direction ofNa^+-Ca^(2+) exchange over myocardial sarcolemma (SL), thus a large quantity of extracellularCa^(2+) fluxed over the SL to the intracellular space, forming a condition of intracellular Ca^(2+)overload, which leads to irreversible damage of the myocardium.展开更多
Objective Background and Objects: Naturally occurring temporal variability of action potentialduration (APD) in isolated myocytes has been noted. Most of the studies have beenfocusing on analyzes of the differences in...Objective Background and Objects: Naturally occurring temporal variability of action potentialduration (APD) in isolated myocytes has been noted. Most of the studies have beenfocusing on analyzes of the differences in ionic channels and currents among theepicardial-, mid-myocardial-(M) and endocardial myocytes, and the rate-dependent (adaptation) characteristics of APD. We have found that the change in APD during achange in frequency of stimulation mostly reflects a change in rate of repolarization at distinct membrane potential levels. We assumed that in the myocytes, there is balancing mechanism, which is constantly adjusting the various ionic currents accommodating to the changing conditions. This intrinsic ability of adaptation is important and may offer some of the consequences of the transmural heterogeneity in adaptation of APD. This adaptive behaviors maybe equally important in maintaining the normal electrophysiological properties and in induction of arrhythmia in a case of error in normal adaptation. Though most studies of Na +/Ca 2+ exchange (NCX) has been emphasized on its reverse activaty during pathyological condition. Our hypothesis is that reverse activaty of NCX also plays an important role in adjusting the repolarization of AP during a physiological condition. A mismatch between action potential (AP) repolarization and relaxation of the contraction can be caused by intracellular Ca 2+ transport abnormalities. Ca 2+ influx via reverse activation of NCX can load the sarcoplasmic recticulum (SR), which has arrhythmogenic effect.Methods We studied the single myocytes from the left ventricle of adult mongrel dogs. During the cell separation, collagenase was perfused through LAD by Langandorff system. We use the patch-clamp system to determinded AP in current clamp mode. Myocyte contraction was imaged by a video camera, shortening of unloaded myocytes was detected by a video edge motion detector, using changes in light intensity at the edges of the myocyte. Results From 60 consecutive recorded APs at 展开更多
文摘Whether La3+ can enter human peripheral blood lymphocytes by the Na+/Ca2+ exchanger or not and the effect of La3+ on the Na+/Ca2+ exchanger activity are examined by fura-2 technique. And that whether La3+ is sequestered by intracellular organelles (mainly endoplasmic reticulum and mitochondria) is studied by this method. La3+uptake is obviously stimulated by pretreating the cells with ouabain and by removing extracellular Na+, and intracellular La3+concentration ([La3+]i) is directly proportional to its extracellular concentration ([La3+]o). But when [La3+]o exceeds 0.4 mmol/L, the 340/380 nm ratio of fluorescence is no longer varied and the maximum [La3+]i is 1.5X10-12 mol · L-1. The higher concentration of La3+ (0.1 mmol/L) increases Na+/Ca2+ exchange-mediated calcium influx, but lower concentration (10 μmol/L) appears to block calcium influx. The results also suggest that cytosolic La3+ is transported by the ATP-dependent Ca2+ pump. Intracellular Ca2+ stores are depleted by ionomycin, and then ionomycin is added again during the period of La3+ uptake, the 340/380 nm ratio of fluorescence is also increased, these results indicate that La3+ is sequestered by intracellular organelles. A characterization of fura-2-La3+ interaction in solution simulating intracellular ionic composition (pH 7.05) shows that La3+forms a 1: 1 fura-2-La3+complex, and the apparent dissociation constant of La3+ for fura-2 (Kd) is 1.7*10-12 mol · L-1. In addition, the limit of detection of fura-2 for La3+ and Ca2+ is 10-12 and 10-8 mol · L-1 respectively.
文摘Previously we have shown that 4 Hz and 8 Hz EMF exposures have depressing effect on the thermodynamic activity of water, which decreases peroxide formation. It has also been shown that 4 Hz EMF-treated physiological solution modulates the growth and development of microbes and heart muscle contractility, but 8 Hz EMF has pronounced inhibitory effect on bacterial growth and development. Therefore, in order to elucidate the possible mechanism of 4 Hz and 8 Hz EMF effects on heart muscle function, in the present work the effects of 4 Hz and 8 Hz EMF exposures on heart muscle tissue hydration, the sensitivity of 4 Hz and 8 Hz EMF-induced tissue hydration to 10−4 M ouabain (Na+/K+ pump inhibition) and 10−9 M ouabain (activation of intracellular signaling system) as well as the effects of 4 Hz and 8 Hz EMF exposures on the number of Na+/K+ pump units in the membrane of both young and old rats have been studied. The obtained data allow us to suggest that 8 Hz EMF exposure has more pronounced age-dependent modulation effect on tissue hydration of heart muscle than 4 Hz EMF and this effect is sensitive to Na+/K+ pump activity and intracellular signaling system.
基金"7-5 project" supported by the National Research Foundation of China.
文摘Results from a systematic experiment on isolated perfused rat heart and isolated myc-cytes of adult rat showed that the mechanism of calcium influx during myocardial ischemia-reperfusion is due to the development of intracellular sodium overload during ischemic pe-riod, on reperfusion, the high intracellular Na^+ content activated the reverse direction ofNa^+-Ca^(2+) exchange over myocardial sarcolemma (SL), thus a large quantity of extracellularCa^(2+) fluxed over the SL to the intracellular space, forming a condition of intracellular Ca^(2+)overload, which leads to irreversible damage of the myocardium.
文摘Objective Background and Objects: Naturally occurring temporal variability of action potentialduration (APD) in isolated myocytes has been noted. Most of the studies have beenfocusing on analyzes of the differences in ionic channels and currents among theepicardial-, mid-myocardial-(M) and endocardial myocytes, and the rate-dependent (adaptation) characteristics of APD. We have found that the change in APD during achange in frequency of stimulation mostly reflects a change in rate of repolarization at distinct membrane potential levels. We assumed that in the myocytes, there is balancing mechanism, which is constantly adjusting the various ionic currents accommodating to the changing conditions. This intrinsic ability of adaptation is important and may offer some of the consequences of the transmural heterogeneity in adaptation of APD. This adaptive behaviors maybe equally important in maintaining the normal electrophysiological properties and in induction of arrhythmia in a case of error in normal adaptation. Though most studies of Na +/Ca 2+ exchange (NCX) has been emphasized on its reverse activaty during pathyological condition. Our hypothesis is that reverse activaty of NCX also plays an important role in adjusting the repolarization of AP during a physiological condition. A mismatch between action potential (AP) repolarization and relaxation of the contraction can be caused by intracellular Ca 2+ transport abnormalities. Ca 2+ influx via reverse activation of NCX can load the sarcoplasmic recticulum (SR), which has arrhythmogenic effect.Methods We studied the single myocytes from the left ventricle of adult mongrel dogs. During the cell separation, collagenase was perfused through LAD by Langandorff system. We use the patch-clamp system to determinded AP in current clamp mode. Myocyte contraction was imaged by a video camera, shortening of unloaded myocytes was detected by a video edge motion detector, using changes in light intensity at the edges of the myocyte. Results From 60 consecutive recorded APs at
