We have studied transmembrane La3+ movement in rat ventricular myocytes for the first time by using the whole-cell patch-clamp recording mode. La3+ (0.01-5.0 mmol/L) could not bring out inward currents through the L-t...We have studied transmembrane La3+ movement in rat ventricular myocytes for the first time by using the whole-cell patch-clamp recording mode. La3+ (0.01-5.0 mmol/L) could not bring out inward currents through the L-type calcium channel in rat ventricular myocytes, while it could enter the cells by the same way carried by 1μmo1/L ionomycin. When the outward Na+ concentration gradient is formed, La3+ can enter the cells via Na-Ca exchange, and the exchange currents increase with the increase of external La3+ concentrations. But compared with Na-Ca exchange currents in the same concentration, the former is only 14%-38% of the latter. The patch-clamp experiment indicates that La3+ normally can not enter ventricular myocytes through L-type calcium channel, but it can enter the cells via Na-Ca exchange.展开更多
Previously we have shown that nM ouabain-induced activation of cAMP-dependent Na/Ca exchange in reverse (R) mode in cell membrane has age-dependent weakening hydration effect on heart muscle and brain tissues and such...Previously we have shown that nM ouabain-induced activation of cAMP-dependent Na/Ca exchange in reverse (R) mode in cell membrane has age-dependent weakening hydration effect on heart muscle and brain tissues and such Na/Ca exchange is characterized by quantum mechanical sensitivity. As in biological experiments radioactive 45Ca is used for the study of cold 40Ca exchange in cells and organisms, in the present work, the age-dependent effect of physiological solution (PS) containing either 40Ca or 45Ca on tissue hydration in different experimental conditions was studied in order to evaluate the bioequivalence of these two forms of Ca. The obtained data indicate that the intraperitoneal injections of 40Ca PS and 45Ca PS leading to activation of RNa/40Ca and RNa/45Ca exchanges, respectively, have different age-dependent effects on heart muscle and brain tissue hydration. As in myocyte membrane, the Na/Ca exchange is more expressed than in neuronal membrane, the age-dependent heart muscle hydration is more sensitive to quantum properties of Ca than brain tissue hydration. The [45Ca]i, in contrary to [40Ca]i, has age-dependent weakening and stabilizing effect on tissue hydration and makes the latter insensitive to ouabain. The obtained data bring us to a strong conclusion that RNa/Ca exchange has quantum mechanical properties and in biological experiments radioactive 45Ca cannot be used for adequate estimation of the functional activity of 40Ca ions in cells and organisms.展开更多
While delayed structural alterations of muscles(DSAM) induced by strenuous exercises have always been one of the commoest findings in sports-related muscle injuries, its pathogenesis remains unidentified. And although...While delayed structural alterations of muscles(DSAM) induced by strenuous exercises have always been one of the commoest findings in sports-related muscle injuries, its pathogenesis remains unidentified. And although acupuncture at Ashi points has proved its efficacy in halting the progress of DASM and speeding up its recovery, we know very little about the underlying mechanism. This paper presents our work on these problems by using amphibian DSAM models created by electrostimulation, and such research tools as electromicroscopy, enzymology, the fluores cence indicator, Fura 2, and electron probe microanalysis (EPMA). Our results indicate:1) As revealed by EPMA, cytoplasmic Ca continued to rbe,reaching 3.07 and 5. 33 mmol/kg dry wt. 3 and 6hrs afterelectrostimlation respectively. Analysis of regions with variousstructural alterations showed rise of Ca concentration inparallel with severity of myofibrillar injury. Testing with Fura 2also demonstrated obvious increase of free Ca++ in cytoplasm 3 hrs after electrostimulation. All these showed a positive correlation between DSAM and the increase of intracellular Ca++.2) Further study to explore the possible mechanisms underlying the increase of cytoplasmic Ca++ revealed two sources. During the initial rise of cytoplasmic Ca, a decline of sarcoplasmic reticulum Ca content was found, suggesting the contribution of sarcoplasmic reticulum Ca. However, no further decline was noted despite continued rise of cytoplasmic Ca, which, then, could only be accounted for by extracellular contribution.3) In skeletal muscles after long-term exhaustive stimulation and in those with delayed structural alterations,the inereased cytoplasmic Ca was quickly lowered by acupuncture, down to lpretest level 10 minutes after acupuncture. Meanwhile,no marked change in sarcoplasmic reticulum Ca content and in enzymic activity of Ca, Mg-ATPase was found, indicating no contribution on their part to the quick lowering of cytoplasmic Ca. But a rapid rise of cytoplasmic Na was found. And addition o展开更多
At present, there are relevant scientific materials on the cellular and molecular mechanisms of electrogenic Na/K pump function and structure, as well as on the potential- and ligand-activated ionic channels in the me...At present, there are relevant scientific materials on the cellular and molecular mechanisms of electrogenic Na/K pump function and structure, as well as on the potential- and ligand-activated ionic channels in the membrane. However, the role of electrogenic Na/K pump in regulation of semipermeable properties of cell membrane has not been elucidated yet, which is due to the fact that our knowledge about the biophysical properties of cell membrane is based on the conductive membrane theory of Hodgkin-Huxley-Katz, which is developed on internally perfused squid axon and lacks intracellular metabolism. Thus, the accumulated abundance of data on the role of G-proteins-dependent intracellular signaling system in regulation of Na/K pump activity and biophysical properties of cell membrane presumes fundamental revision of some statements of membrane theory. The aim of the present review is to briefly demonstrate our and literature data on cell hydration-induced auto-regulation of Na/K pump as well as on its role in metabolic control of semipermeable properties and excitability of neuronal membrane, which are omitted in the study of internally perfused squid axon.展开更多
Objective To study the alteration of Na + Ca 2+ exchange in rat cardiac sarcolemmal membrane during phases of septic shock Methods Sepsis was induced by cecal ligation and puncture (CLP) Na + Ca 2+ ...Objective To study the alteration of Na + Ca 2+ exchange in rat cardiac sarcolemmal membrane during phases of septic shock Methods Sepsis was induced by cecal ligation and puncture (CLP) Na + Ca 2+ exchange was assayed by radioactive analysis Results Na + dependent 45 Ca 2+ uptake was decreased by 62%-69% in late phase of sepsis, whereas it was not affected in early phase of sepsis Na + Ca 2+ exchange stimulated by 5'guanylyl imidodiphosphate [Gpp(NH)p] was decreased by 65 7% in late phase of sepsis but unaltered in early phase of sepsis Two agonists (angiotensin Ⅱ and phenylephrine) coupled to Gq and a protein kinase C activator, phorbol 12 myristate 13 acetate (PMA) all inhibited Na + Ca 2+ exchange in late phase of sepsis Na + Ca 2+ exchange activities induced by phosphorylation of Na + Ca 2+ exchange were decreased in late phase of sepsis, whereas inhibition of Na + Ca 2+ exchange by dephosphorylation was increased both in early and late phases of sepsis Conclusion The alteration of Na + Ca 2+ exchange during different phases of sepsis might be related to the activities of Gq, protein kinase C, and phosphorylation/dephosphorylation展开更多
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
基金This work was supported by the National Natural Science Foundation of China (Grant No. 29890280).
文摘We have studied transmembrane La3+ movement in rat ventricular myocytes for the first time by using the whole-cell patch-clamp recording mode. La3+ (0.01-5.0 mmol/L) could not bring out inward currents through the L-type calcium channel in rat ventricular myocytes, while it could enter the cells by the same way carried by 1μmo1/L ionomycin. When the outward Na+ concentration gradient is formed, La3+ can enter the cells via Na-Ca exchange, and the exchange currents increase with the increase of external La3+ concentrations. But compared with Na-Ca exchange currents in the same concentration, the former is only 14%-38% of the latter. The patch-clamp experiment indicates that La3+ normally can not enter ventricular myocytes through L-type calcium channel, but it can enter the cells via Na-Ca exchange.
文摘Previously we have shown that nM ouabain-induced activation of cAMP-dependent Na/Ca exchange in reverse (R) mode in cell membrane has age-dependent weakening hydration effect on heart muscle and brain tissues and such Na/Ca exchange is characterized by quantum mechanical sensitivity. As in biological experiments radioactive 45Ca is used for the study of cold 40Ca exchange in cells and organisms, in the present work, the age-dependent effect of physiological solution (PS) containing either 40Ca or 45Ca on tissue hydration in different experimental conditions was studied in order to evaluate the bioequivalence of these two forms of Ca. The obtained data indicate that the intraperitoneal injections of 40Ca PS and 45Ca PS leading to activation of RNa/40Ca and RNa/45Ca exchanges, respectively, have different age-dependent effects on heart muscle and brain tissue hydration. As in myocyte membrane, the Na/Ca exchange is more expressed than in neuronal membrane, the age-dependent heart muscle hydration is more sensitive to quantum properties of Ca than brain tissue hydration. The [45Ca]i, in contrary to [40Ca]i, has age-dependent weakening and stabilizing effect on tissue hydration and makes the latter insensitive to ouabain. The obtained data bring us to a strong conclusion that RNa/Ca exchange has quantum mechanical properties and in biological experiments radioactive 45Ca cannot be used for adequate estimation of the functional activity of 40Ca ions in cells and organisms.
文摘While delayed structural alterations of muscles(DSAM) induced by strenuous exercises have always been one of the commoest findings in sports-related muscle injuries, its pathogenesis remains unidentified. And although acupuncture at Ashi points has proved its efficacy in halting the progress of DASM and speeding up its recovery, we know very little about the underlying mechanism. This paper presents our work on these problems by using amphibian DSAM models created by electrostimulation, and such research tools as electromicroscopy, enzymology, the fluores cence indicator, Fura 2, and electron probe microanalysis (EPMA). Our results indicate:1) As revealed by EPMA, cytoplasmic Ca continued to rbe,reaching 3.07 and 5. 33 mmol/kg dry wt. 3 and 6hrs afterelectrostimlation respectively. Analysis of regions with variousstructural alterations showed rise of Ca concentration inparallel with severity of myofibrillar injury. Testing with Fura 2also demonstrated obvious increase of free Ca++ in cytoplasm 3 hrs after electrostimulation. All these showed a positive correlation between DSAM and the increase of intracellular Ca++.2) Further study to explore the possible mechanisms underlying the increase of cytoplasmic Ca++ revealed two sources. During the initial rise of cytoplasmic Ca, a decline of sarcoplasmic reticulum Ca content was found, suggesting the contribution of sarcoplasmic reticulum Ca. However, no further decline was noted despite continued rise of cytoplasmic Ca, which, then, could only be accounted for by extracellular contribution.3) In skeletal muscles after long-term exhaustive stimulation and in those with delayed structural alterations,the inereased cytoplasmic Ca was quickly lowered by acupuncture, down to lpretest level 10 minutes after acupuncture. Meanwhile,no marked change in sarcoplasmic reticulum Ca content and in enzymic activity of Ca, Mg-ATPase was found, indicating no contribution on their part to the quick lowering of cytoplasmic Ca. But a rapid rise of cytoplasmic Na was found. And addition o
文摘At present, there are relevant scientific materials on the cellular and molecular mechanisms of electrogenic Na/K pump function and structure, as well as on the potential- and ligand-activated ionic channels in the membrane. However, the role of electrogenic Na/K pump in regulation of semipermeable properties of cell membrane has not been elucidated yet, which is due to the fact that our knowledge about the biophysical properties of cell membrane is based on the conductive membrane theory of Hodgkin-Huxley-Katz, which is developed on internally perfused squid axon and lacks intracellular metabolism. Thus, the accumulated abundance of data on the role of G-proteins-dependent intracellular signaling system in regulation of Na/K pump activity and biophysical properties of cell membrane presumes fundamental revision of some statements of membrane theory. The aim of the present review is to briefly demonstrate our and literature data on cell hydration-induced auto-regulation of Na/K pump as well as on its role in metabolic control of semipermeable properties and excitability of neuronal membrane, which are omitted in the study of internally perfused squid axon.
文摘Objective To study the alteration of Na + Ca 2+ exchange in rat cardiac sarcolemmal membrane during phases of septic shock Methods Sepsis was induced by cecal ligation and puncture (CLP) Na + Ca 2+ exchange was assayed by radioactive analysis Results Na + dependent 45 Ca 2+ uptake was decreased by 62%-69% in late phase of sepsis, whereas it was not affected in early phase of sepsis Na + Ca 2+ exchange stimulated by 5'guanylyl imidodiphosphate [Gpp(NH)p] was decreased by 65 7% in late phase of sepsis but unaltered in early phase of sepsis Two agonists (angiotensin Ⅱ and phenylephrine) coupled to Gq and a protein kinase C activator, phorbol 12 myristate 13 acetate (PMA) all inhibited Na + Ca 2+ exchange in late phase of sepsis Na + Ca 2+ exchange activities induced by phosphorylation of Na + Ca 2+ exchange were decreased in late phase of sepsis, whereas inhibition of Na + Ca 2+ exchange by dephosphorylation was increased both in early and late phases of sepsis Conclusion The alteration of Na + Ca 2+ exchange during different phases of sepsis might be related to the activities of Gq, protein kinase C, and phosphorylation/dephosphorylation
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.
