首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 78 毫秒
1.
Brain (31)P-neurometabolites play an important role in energy and membrane metabolism. Unambiguous identification and quantification of these neurochemicals in different brain regions would be a great aid in advancing the understanding of metabolic processes in the nervous system. Phosphomonoester (PME), consisting of phosphoethanolamine (PE) and phosphocholine (PC), is the "building block" for membranes, while phosphodiesters (PDE), consisting of glycerophosphocholine (GPC) and glycerophosphoethanolamine (GPE) metabolites are involved in the membrane breakdown process. In the clinical setting, generating well-resolved spectra for PC, PE, GPC, and GPE could be crucial phospholipids in providing information regarding membrane metabolism. We present here a new experimental approach for generating well-resolved (31)P spectra for PC and PE as well as for GPC, GPE, and other (31)P metabolites. Our results (based on uni-dimensional (1D) and multi-voxel (31)P studies) indicate that an intermediate excitation pulse angle (35°) is best suited to obtain well-resolved PC/PE and GPC/GPE resonance peaks. Our novel signal processing scheme allows generating metabolite maps of different phospholipids include PC/PE and GPC/GPE using the 'time-domain-frequency-domain' method as referred to in the MATLAB programming language.  相似文献   

2.
Brain 31P-neurometabolites play an important role in energy and membrane metabolism. Unambiguous identification and quantification of these neurochemicals in different brain regions would be a great aid in advancing the understanding of metabolic processes in the nervous system. Phosphomonoester (PME), consisting of phosphoethanolamine (PE) and phosphocholine (PC), is the “building block” for membranes, while phosphodiesters (PDE), consisting of glycerophosphocholine (GPC) and glycerophosphoethanolamine (GPE) metabolites are involved in the membrane breakdown process. In the clinical setting, generating well-resolved spectra for PC, PE, GPC, and GPE could be crucial phospholipids in providing information regarding membrane metabolism. We present here a new experimental approach for generating well-resolved 31P spectra for PC and PE as well as for GPC, GPE, and other 31P metabolites. Our results (based on uni-dimensional (1D) and multi-voxel 31P studies) indicate that an intermediate excitation pulse angle (35°) is best suited to obtain well-resolved PC/PE and GPC/GPE resonance peaks. Our novel signal processing scheme allows generating metabolite maps of different phospholipids include PC/PE and GPC/GPE using the ‘time-domain–frequency-domain’ method as referred to in the MATLAB programming language.  相似文献   

3.
H C Liu  C W Chi  T Y Liu  L H Liu  W M Luh  C H Hsieh  W G Wu 《Life sciences》1991,48(21):2057-2063
Changes of phosphate metabolism in brains of neonate, weaning and adult rats were compared using both in vivo and in vitro nuclear magnetic resonance spectra. Ratios of phosphocreatine/nucleoside triphosphate (PCr/NTP) were the same in neonatal brain in both in vivo and in vitro studies, but not in weaning and adult brains. This discrepancy may have resulted from extended cerebral hypoxia due to slowed freezing of the brain by the increased skull thickness and brain mass in the weaning and adult rats. Variations in in vitro extraction condition for this age-related study may lead to systematic errors in the adult rats. Nevertheless, the phosphomonoester/nucleoside triphosphate (PME/NTP) ratios in extracts of brain from neonatal rats were higher than those obtained in vivo. In addition, the glycerophosphorylethanolamine plus glycerophosphorylcholine/nucleoside triphosphate (GPE+GPC/NTP) ratios, which were not measurable in vivo, showed age-dependent increase in extracts of rat brain. Some of the phosphomonoester and phosphodiester molecules in rat brain may be undetectable in in vivo NMR analysis because of their interaction with cellular components. The total in vitro GPE and GPC concentration in brain from neonatal rat was estimated to be 0.34 mmole/g wet tissue.  相似文献   

4.
Phospholipid metabolism in cancer cells monitored by 31P NMR spectroscopy   总被引:5,自引:0,他引:5  
Addition of choline, ethanolamine, or hemicholinium-3 (a choline kinase inhibitor) to the perfusate of human breast cancer cells monitored by 31P NMR spectroscopy resulted in significant changes to phosphomonoester (PME) and phosphodiester (PDE) signals. These results enable us to assign the PMEs to phosphcholine (PC) and phosphoethanolamine (PE), the PDEs to glycerophosphorylcholine and glycerophosphorylethanolamine, and to define the pathways producing them. The PMEs are products of choline and ethanolamine kinases, the first steps in phospholipid synthesis; and the PDEs are substrates of glycerophosphorylcholine phosphodiesterase, the last step in phospholipid catabolism. Furthermore, PC and PE peaks are twice as intense in cells at log phase versus confluency. We also observed these signals in vivo in human colon and breast tumors grown in mice. Since PMEs are low in most nonproliferating tissues, they could form a basis for noninvasive diagnosis. Also, PE and PC are situated between the control enzymes of two major synthetic pathways and will allow noninvasive 31P NMR studies of these pathways in intact cells and in vivo.  相似文献   

5.
31P NMR spectra of tumor-infiltrating lymphocytes (TILs) were found to be significantly different from those of normal peripheral lymphocytes. The greatest difference was in the phosphodiester (PDE) region, mainly in the glycerophosphocholine (GPC) signal. Short-term activation of peripheral lymphocytes with interleukin-2 induced a small increase in ATP levels. In all lymphocytes the phosphomonoester (PME) region is dominated by phosphoethanolamine (PE), while there is an unusual absence of phosphocholine (PC). Perfusion of these cells with high concentrations of choline caused only a minimal increase in PC, indicating that choline kinase is not the rate limiting step of lecithin synthesis in lymphocytes.  相似文献   

6.
Spectral changes in human hepatic tumours and possible systemic effects of tumour on host liver were assessed by 31P amnd 1H in vitro NMR spectroscopy. The 1H and 31P spectra from liver tumour biopsies showed significant elevation in phosphoethanolamine, phosphocholine, taurine, citrate, alanine, lactate and glycine, and significant reduction in GPE (glycerophosphoethanolamine), GPC (glycerophosphocholine), creatine and threonine compared to histologically normal tissue. 31P-NMR spectra obtained from histologically normal tissue within tumour-bearing livers showed significant elevation in phosphoethanolamine and phosphocholine compared to data from liver biopsies from nontumour-bearing patients (pancreatitis). These results suggest that alterations in membrane metabolism in host liver can be detected by 31P-NMR.  相似文献   

7.

Purpose

To assess the ability of a polarization transfer (PT) magnetic resonance spectroscopy (MRS) technique to improve the detection of the individual phospholipid metabolites phosphocholine (PC), phosphoethanolamine (PE), glycerophosphocholine (GPC), and glycerophosphoethanolamine (GPE) in vivo in breast tumor xenografts.

Materials and Methods

The adiabatic version of refocused insensitive nuclei enhanced by polarization transfer (BINEPT) MRS was tested at 9.4 Tesla in phantoms and animal models. BINEPT and pulse-acquire (PA) 31P MRS was acquired consecutively from the same orthotopic MCF-7 (n = 10) and MDA-MB-231 (n = 10) breast tumor xenografts. After in vivo MRS measurements, animals were euthanized, tumors were extracted and high resolution (HR)-MRS was performed. Signal to noise ratios (SNRs) and metabolite ratios were compared for BINEPT and PA MRS, and were also measured and compared with that from HR-MRS.

Results

BINEPT exclusively detected metabolites with 1H-31P coupling such as PC, PE, GPC, and GPE, thereby creating a significantly improved, flat baseline because overlapping resonances from immobile and partly mobile phospholipids were removed without loss of sensitivity. GPE and GPC were more accurately detected by BINEPT in vivo, which enabled a reliable quantification of metabolite ratios such as PE/GPE and PC/GPC, which are important markers of tumor aggressiveness and treatment response.

Conclusion

BINEPT is advantageous over PA for detecting and quantifying the individual phospholipid metabolites PC, PE, GPC, and GPE in vivo at high magnetic field strength. As BINEPT can be used clinically, alterations in these phospholipid metabolites can be assessed in vivo for cancer diagnosis and treatment monitoring.  相似文献   

8.
Significant amounts of phospholipid N-methyltransferase activity in murine thymocytes were found to be distributed on the plasma membrane. The enzyme activity had an optimum pH of 9. The presence of divalent cations, Mg2+ (10 mM) or Ca2+ (1 mM), and EGTA separately in the assay had only a small effect on the enzyme activity. However, addition of both 10 mM Mg2+ and 1 mM Ca2+ increased the enzyme activity. The presence of two enzymes for each conversion of phosphatidylethanolamine (PE) to phosphatidylmonomethylethanolamine (PME) and PME to phosphatidylcholine (PC) was suggested by the result of the determination of the incorporated radioactivity into PME, phosphatidyldimethylethanolamine (PDE) and PC; the apparent Km values for S-adenosyl-L-methionine were 20 and 400-500 microM for the conversion of PE to PME and for the conversion of PME to PC they were 5 microM and 40 microM. S-Adenosyl-L-homocysteine (AdoHcy), a known inhibitor of enzymatic methylation, competitively inhibited [14C]methyl incorporation into total lipid. The apparent Ki value for AdoHcy was 44.7 microM. Two phospholipid N-methyltransferases were partially purified by extraction with sodium deoxycholate, gel filtration on Sephadex G-75, and affinity column chromatography on AdoHcy-Sepharose. One enzyme, mainly catalyzing the formation of PME, was purified approximately 1548-fold and the other catalyzing the formation of PDE and PC, was purified approximately 629- to 703-fold. However, the former still contained a little activity for PDE and PC formation and the latter contained a little activity for PME formation. In these partially purified phospholipid N-methyltransferase preparations, little contaminating protein O-carboxylmethyltransferase activity was observed; however, significant PC-phospholipase A2 activity was detected. This result may suggest that phospholipid N-methyltransferases associate with phospholipase A2 in the thymocyte plasma membrane.  相似文献   

9.
Both in vivo and in vitro 31P-NMR spectroscopy were used to demonstrate metabolic changes in rat liver as a function of time after exposure to either carbon tetrachloride (CCl4) or bromotrichloromethane (BrCCl3). The inorganic phosphate resonance, measured in vivo, moves upfield, which is associated with a decrease in cytosolic pH over a 12 or 20 h period (for BrCCl3 or CCl4, respectively). Intoxication by CCl4 or BrCCl3 causes an intracellular acidosis to pH 7.05 or 6.82 (+/- 0.05), respectively. Also, it has been found that halocarbon exposure increases the amounts of phosphomonoesters (PME) detected. High resolution in vitro 31P-NMR spectroscopy studies of perchloric acid extracts of CCl4-treated rat livers indicated a significant increase in the height of the phosphocholine resonance in the PME region 4-5 h after CCl4 exposure.  相似文献   

10.
Levels of phosphomonoester (PME), phosphodiester (PDE), ATP and pH in brain cells of chicken embryos rotated for 24 h in a clinostat during the period of hatching the 13th day (E13) and 15th day (E15) embryos were investigated by using 31P-NMR spectroscopy. Significant increases in the values of PME, ATP and pH were identified after E13 rotating for 24 h. With the same treatment, differences were obtained in the phospholipid and energy metabolism of E15, but no significant levels have been reached . The calorimetric assay (malachite green method) was used for measuring the activity of total ATPase. A dramatic decrease was evident in the activity of ATPase in brain cells of rotated E13 and E15. The former is more sensitive than the latter. All the levels mentioned above could restore in 24 h after the rotation stopped, except that the level of ATP was still higher than the control.  相似文献   

11.
Levels of phosphomonoester (PME), phosphodiester (PDE), ATP and pH in brain cells of chicken embryos rotated for 24 h in a clinostat during the period of hatching the 13th day (E13) and 15th day (E15) embryos were investigated by using 31P-NMR spectroscopy. Significant increases in the values of PME, ATP and pH were identified after E13 rotating for 24 h. With the same treatment, differences were obtained in the phospholipid and energy metabolism of E15, but no significant levels have been reached . The calorimetric assay (malachite green method) was used for measuring the activity of total ATPase. A dramatic decrease was evident in the activity of ATPase in brain cells of rotated E13 and E15. The former is more sensitive than the latter. All the levels mentioned above could restore in 24 h after the rotation stopped, except that the level of ATP was still higher than the control.  相似文献   

12.
Levels of phosphomonoester (PME), phosphodiester (PDE), ATP and pH in brain cells of chicken embryos rotated for 24 h in a clinostat during the period of hatching the 13th day (E13) and 15th day (E15) embryos were investigated by using31P-NMR spectroscopy. Significant increases in the values of PME, ATP and pH were identified after E13 rotating for 24 h. With the same treatment, differences were obtained in the phospholipid and energy metabolism of E15, but no significant levels have been reached. The calorimetric assay (malachite green method) was used for measuring the activity of total ATPase. A dramatic decrease was evident in the activity of ATPase in brain cells of rotated E13 and E15. The former is more sensitive than the latter. All the levels mentioned above could restore in 24 h after the rotation stopped, except that the level of ATP was still higher than the control.  相似文献   

13.
31P-NMR has been used extensively for the study of cytosolic small molecule phosphates in vivo and phospholipid structures in vitro. We present in this paper a series of studies of the brain by 31P-NMR, both in vivo and in extracts, showing the information that can be derived about phospholipids. 31P-NMR spectra of mouse brain at 73 mHz are characterised by almost a complete absence of the large phosphodiester peak in comparison to equivalent spectra at 32 mHz. Proton decoupled spectra in vivo, and spectra of extracts, show that the phosphodiester peak observed in 32 mHz spectra in vivo is mainly due to phospholipid bilayers. Homogenates of quaking and control mouse brains, and of bovine grey matter, show another narrower phosphodiester peak possibly from small phospholipid vesicles. This peak is increased in intensity in the affected mice. These experiments demonstrate the presence of three major components contributing to the phosphodiester resonance: bilayer phospholipids, more mobile phospholipids, and the freely soluble cytosolic molecules glycerophosphocholine and glycerophosphoethanolamine. These NMR methods for non-invasive investigation of phospholipid structures in the brain might be extended to studies of patients with membrane involved diseases such as multiple sclerosis.  相似文献   

14.
Recent articles have confused the steady state concentration of radioactivity in N-methylphosphatidylethanolamine (PME) and N,N-dimethylphosphatidylethanolamine (PDE) with the amount of these products formed during the conversion of phosphatidylethanolamine (PE) to phosphatidylcholine (PC). This paper clarifies this problem and reports the apparent Km values for AdoMet and pH optima for the conversion of PE to PME, PDE, and PC by rat liver microsomes. We purified AdoMet and [methyl-3H]AdoMet and measured the transfer of tritium to PME, PDE, and PC as a function of time. There was an initial lag in the formation of [3H]PC followed by linear incorporation of isotope. In contrast, labeled PME and PDE reached and maintained steady state levels within 1 to 2 min. Hence, calculations of the rate of formation of PME, PDE, and PC must take into account the subsequent conversion of PME and PDE to PC. The PE N-methyltransferase was assayed at pH 6.6, 9.2, and 10.25 and the apparent Km for AdoMet for the three methylation reactions was calculated. The formation of PME was best estimated by the dpm in PME + 1/2 dpm in PDE + 1/3 dpm in PC. The synthesis of PDE from PME was estimated from 1/2 dpm in PDE and 1/3 dpm in PC, and the formation of PC from PDE estimated by 1/3 dpm in PC. The apparent Km for AdoMet at pH 10.25 for the conversion of PE to PME was 58 microM, PME to PDE was 65 microM, and PDE to PC was 96 microM. The pH optimum for each of these methylation reactions was 10.25. This high value was not due to alkaline degradation of AdoMet or denaturation of the enzyme. The apparent Km for AdoMet was also estimated for the conversion of exogenous PME to PDE (50 microM) and exogenous PDE to PC (45 microM). Since recent studies on the methylation of PE have not taken into account the conversion of newly formed PME and PDE to PC, the results and conclusions about apparent Km values for AdoMet, pH optima, and the number of enzymes involved must be re-evaluated.  相似文献   

15.
Size-dependent changes in therapeutically relevant and interrelated metabolic parameters of a murine fibrosarcoma (FSaII) were investigated in vivo using conscious (unanesthetized) animals and tumor sizes less than or equal to 2% of body weight. Tumor pH and bioenergetics were evaluated by 31P nuclear magnetic resonance spectroscopy (31P-MRS), and tumor tissue oxygen tension (pO2) distribution was examined using O2-sensitive needle electrodes. During growth FSaII tumors showed a progressive loss of phosphocreatine (PCr) and nucleoside triphosphate (NTP) with increasing inorganic phosphate (Pi) and phosphomonoester (PME) signals. Ratios for PCr/Pi, PME/Pi, NTP/Pi, and phosphodiester/inorganic phosphate (PDE/Pi) as well as pH determined by 31P-NMR (pHNMR) and the mean tissue pO2 progressively declined as the tumors increased in size. The only relevant ratio increasing with tumor growth was PME/NTP. When the mean tissue pO2 value was plotted against pHNMR, NTP/Pi, PCr/Pi, PME/Pi, and PDE/Pi for tumor groups of similar mean volumes, a highly significant positive correlation was observed. There was a negative correlation between mean tumor tissue pO2 values and PME/NTP. From these results we concluded that 31P-MRS can detect changes in tumor bioenergetics brought about by changes in tumor oxygenation. Furthermore, the close correlation between oxygenation and energy status suggests that the microcirculation in FSaII tumors yields an O2-limited energy metabolism. Finally, a correlation between the proportion of pO2 readings between 0 and 2.5 mmHg and the radiobiologically hypoxic cell fraction in FSaII tumors was observed. The latter finding might be of particular importance for radiation therapy.  相似文献   

16.
When soybean oil containing tocopherol acetate was given to rats once a week subcutaneously for 10-12 months, it caused the development of fibrosarcomas at the injection site in 11 of 15 rats. A tumor produced in this manner proved eminently transplantable into other rats. The molecular species of phospholipid subclasses were determined in primary and transplanted tumors. The molecular species composition of the phospholipid subclasses in both types of tumors were similar. The percentages of diacyl and alkylacyl glycerophosphocholine (GPC) were 90-93 and 6-8% of total phosphatidylcholine, respectively. The percentages of diacyl and alkenylacyl glycerophosphoethanolamine (GPE) were 51 and 45%, respectively, of total phosphatidylethanolamine (PE). Diacyl and alkylacyl GPC species containing arachidonic acid (20:4) composed about 15-16 and 37-40% of each subclass, respectively. Diacyl and alkenylacyl GPE species containing 20:4 composed about 38-40 and 56-60% of each subclass, respectively. Disaturated species of diacyl and alkylacyl GPC composed about 22-24 and 13% of each subclass, respectively, whereas these species of PE composed less than 2%. The fatty acid composition of the other tumor phospholipids was analyzed.  相似文献   

17.
The distribution of water-soluble phosphodiesters (WSPDEs) visible by nuclear magnetic resonance (NMR) in some intact tissues of rainbow trout (Oncorhynchus mykiss walbaum) and in perchloric extracts after partial purification was examined by (31)P NMR spectroscopy. The compounds of interest were serine ethanolamine phosphate (SEP), threonine ethanolamine phosphate (TEP), glycerophosphorylcholine (GPC), and glycerophosphorylethanolamine (GPE). TEP and SEP were mostly accumulated in the heart and less accumulated in the kidney of intact trout. After the extraction procedure, two additional minor resonances were visible and identified as GPC and GPE. The liver of trout contained large amounts of GPE. Similar investigations were conducted by (31)P NMR on hearts and kidneys of two elasmobranchs (Scyliorhinus canicula, Raja clavata) and four teleosts (Anguilla anguilla, Sparus auratus, Dicentrarchus labrax, Scophtlhalmus maximus); comparison with the trout data showed striking interspecies differences in the identity of WSPDEs. All teleosts, except eel and turbot, accumulated predominantly TEP. However, in elasmobranchs, first GPC and then GPE were the major compounds. Whatever the studied species, the relative abundances in the heart and kidney were similar. In the last two decades, two hypotheses were proposed to explain the occurrence of high levels of cytosoluble phosphodiesters: these compounds may constitute an index of phospholipid catabolism or a protective mechanism through which phospholipid levels are kept high. To test them and elucidate the role of these compounds in membrane phospholipid regulation in fish, we investigated the effects of two physiological stresses, that is, seawater adaptation and induced myocardial ischemia, on trout cytosolic phosphodiester levels. A 32.5-min ischemic stress caused no effect on SEP and TEP levels. On the contrary, significant osmotic stress induced changes in the PDEs levels: 2 d after transfer from freshwater to seawater or from seawater to freshwater, both tissues displayed a transient decrease of TEP; however, a 2-d stay in seawater after transfer from freshwater caused a rise in SEP concentration, whereas a 2-d stay in freshwater after transfer decreased SEP level. In conclusion, our experiments suggest a relationship between the high levels of cytosoluble phosphodiesters observed in some fish tissues and resistance to stress.  相似文献   

18.
Summary During the stationary growth phase, the phospholipids of Thiobacillus neapolitanus consisted of phosphatidyl glycerol (PG), diphosphatidyl glycerol (DPG), phosphatidyl-N-monomethylethanolamine (PME) and phosphatidyl ethanolamine (PE) in increasing amounts. In general, the phospholipids increased to a maximum concentration during the stationary phase and then decreased in concentration. Individually, PG and PE increased to a maximum in late lag or early exponential phase and then decreased in concentration. DPG and PME increased during the transition between the exponential and the stationary phase and reached a maximum concentration in the stationary phase. In older cultures, a quantitative interconversion between PG and DPG and PE and PME was observed. A lyso-phospholipid compound also appeared in the late stationary phase.The phospholipid composition of the culture supernatant fluid was essentially similar to that of the cells at all stages of growth. No excessive secretion of these products into the medium was observed at any growth stage of the culture.Abbreviations used PG Phosphatidyl glycerol - DPG Diphosphatidyl glycerol - PME Phosphatidyl-N-monomethylethanolamine - PE Phosphatidyl ethanolamine - GPGPG Glycerophosphoryl glycerophosphoryl glycerol - GPG Glycerophosphoryl glycerol - GPE Glycerophosphoryl ethanolamine - GPME Glycerophosphoryl-N-monomethylethanolamine  相似文献   

19.
The metabolic changes occurring during liver regeneration after partial hepatectomy have been followed by examination of the perchloric extracts by means of NMR spectroscopy. Proton spectra show an abrupt decrease of glycogen, glucose and nucleotides, which are essentially ribonucleotides, in the first hours after hepatectomy. Nucleotides begin to store up again in the third day after hepatectomy, while glucose and glycogen storage builds up from the second day. 31P data evidence a sharp drop of phosphomonoesters, i.e. monophosphosugars (including AMP) and phosphocholine, and phosphodiesters, i.e. GPC and GPE, soon after hepatectomy and a recovery of the initial levels approximately at the 60th hour and a further increase of PDE at later times. The NMR findings are in agreement with the biochemical knowledge of the course of liver regeneration.  相似文献   

20.
Rat1-T1 and MR1 spheroids representseparate transformed phenotypes originated from the same ratfibroblasts that differ in three-dimensional (3D) growth kinetics,histological structure, and oxygenation status. In the present study,31P-NMR spectroscopy of perfused spheroid suspensions wasused to investigate cellular energetics relative to 3D growth,development of necrosis, and cell cycle distribution. Both spheroidtypes were characterized by a remarkably low amount of free (inorganic) phosphate (Pi) and a low phosphocreatine peak. The ratio ofnucleoside triphosphate (NTP) to Pi ranged between 1.5 and2.0. Intracellular pH, NTP-to-Pi ratio, and NTP/cellremained constant throughout spheroid growth, being unaffected by theemergence of oxygen deficiency, cell quiescence, and necrosis. However,a 50% decrease in the ratio of the lipid precursors phosphorylcholineand phosphorylethanolamine (PC/PE) was observed with increasingspheroid size and was correlated with an increasedG1/G0 phase cell fraction. In addition, the ratio of the phospholipid degradation productsglycerophosphorylcholine and glycerophosphorylethanolamine(GPC/GPE) increased with spheroid diameter in Rat1-T1 aggregates.We conclude that changes in phospholipid metabolism, rather thanalterations in energy-rich phosphates, reflect cell quiescence inspheroid cultures, because cells in the inner oxygen-deficient zonesseem to adapt their energy metabolism to the environmental conditionsbefore necrotic cell destruction.

  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号