Sidransky H, Verney E, Orenstein J
Department of Pathology, George Washington University Medical Center, Washington, DC 20037, USA.
Am J Physiol Cell Physiol. 2000 Jun;278(6):C1237-45. doi: 10.1152/ajpcell.2000.278.6.C1237.
This study was concerned with the effects of NaCl administered in vivo or added in vitro to isolated nuclei on [(3)H]tryptophan binding to rat hepatic nuclei assayed in vitro. Hypertonic (10.7%) NaCl administered in vivo to rats caused at 10 min a marked decrease in in vitro binding (total and specific) of [(3)H]tryptophan to hepatic nuclei. In vitro incubation of isolated hepatic nuclei, but not of isolated nuclear envelopes, with added NaCl (particularly at 0.125 x 10(-4) M and 0.25 x 10(-4) M) revealed significant inhibition of [(3)H]tryptophan binding. However, isolated hepatic nuclear envelopes prepared after in vitro incubation of isolated nuclei with added NaCl did show inhibition of [(3)H]tryptophan binding (total and specific) compared with controls. Other salts (KCl, MgCl(2), NaHCO(3), NaC(2)H(3)O(2), NaF, or Na(2)SO(4)), at similar concentrations to that of NaCl except for MgCl(2), when added to isolated nuclei did not appreciably inhibit nuclear tryptophan binding. Kinetic studies of in vitro nuclear [(3)H]tryptophan binding in the presence of 0.125 x 10(-4) M NaCl revealed that binding decreased at 0.5 h and continued to 2 h compared with nuclear [(3)H]tryptophan binding with controls (without NaCl addition). The results obtained in vivo in rats and those obtained in vitro with isolated hepatic nuclei revealed NaCl-induced inhibitory effects on [(3)H]tryptophan binding to hepatic nuclei. Although the inhibitory effects were similar under the two different experimental conditions, the mechanism for each may be different in that the NaCl concentration in hepatic cells after administration of NaCl in vivo was appreciably higher than the low levels added in vitro to the isolated hepatic nuclei.
本研究关注体内给予氯化钠或体外向分离的细胞核中添加氯化钠对体外测定的[³H]色氨酸与大鼠肝细胞核结合的影响。体内给予大鼠高渗(10.7%)氯化钠,10分钟时[³H]色氨酸与肝细胞核的体外结合(总量和特异性)显著降低。分离的肝细胞核(而非分离的核膜)与添加的氯化钠(特别是在0.125×10⁻⁴M和0.25×10⁻⁴M时)进行体外孵育,显示出对[³H]色氨酸结合的显著抑制。然而,在分离的细胞核与添加的氯化钠进行体外孵育后制备的分离肝核膜,与对照组相比,确实显示出对[³H]色氨酸结合(总量和特异性)的抑制。其他盐(氯化钾、氯化镁、碳酸氢钠、醋酸钠、氟化钠或硫酸钠),除氯化镁外,当以与氯化钠相似的浓度添加到分离的细胞核中时,并未明显抑制核色氨酸结合。在0.125×10⁻⁴M氯化钠存在下对体外细胞核[³H]色氨酸结合的动力学研究表明,与对照组(不添加氯化钠)的细胞核[³H]色氨酸结合相比,在0.5小时时结合减少,并持续到2小时。在大鼠体内获得的结果以及在体外对分离的肝细胞核获得的结果显示,氯化钠对[³H]色氨酸与肝细胞核的结合具有抑制作用。尽管在两种不同的实验条件下抑制作用相似,但每种情况的机制可能不同,因为体内给予氯化钠后肝细胞中的氯化钠浓度明显高于体外添加到分离的肝细胞核中的低水平。