Objective: To explore the roles of intracellular pH value (pHi) and sodium-hydrogen exchanger isoform-1 (NHE-1) in the mechanism of multidrug resistance of leukemia cells. Methods: Multidrug resistant cell line HL-60 ...Objective: To explore the roles of intracellular pH value (pHi) and sodium-hydrogen exchanger isoform-1 (NHE-1) in the mechanism of multidrug resistance of leukemia cells. Methods: Multidrug resistant cell line HL-60 induced by doxorubicin(DOX) (called as HL-60/DOX cells) and their parent cell line HL-60 were employed as experiment group and control group. The proliferation and chemosensitivity of the cells were studied by MTT assay, and the expression of multidrug resistance protein (MRP) was detected by immol/Lunocytochemistry. Meanwhile, pHi was measured by spectrofluorometery with a fluorescence dye BCECF-AM. Based on the pHi recovery curve after intracellular acid loading, the activity of NHE-1 was analyzed. The expression of NHE-1 mRNA and MRP mRNA were determined by semi-quantitative RT-PCR. Cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and apoptotic DNA was extracted and electrophoresed. Results: ① The IC 50 values for DOX, MTZ, VCR and homoharringtonine(HT), in HL-60/DOX cells were significantly higher than those in HL-60 cells (P<0.01). HL-60/DOX cells expressed abundant MRP, but HL-60 cells did not. ② pHi of HL-60/DOX cells were significantly higher than that of HL-60 cells(P<0.001). The expression and activity of NHE-1 in HL-60/DOX cells were significantly stronger than those of HL-60 cells. ③After administration of the specific NHE-1 inhibitor dimethyl amiloride (DMA) at a certain range of concentrations, compared with HL-60 cells, the rate of growth inhibition of HL-60/DOX cells increased significantly (P<0.05), the drug-sensitivity of HL-60/DOX cells was significantly sensitive (P<0.01), the expression of MRP and MRP mRNA decreased significantly (P<0.01), the apoptosis rate increased significantly (P<0.01). Conclusion: NHE-1 is involved in the drug-resistant mechanisms of multidrug-resistant HL-60 cells induced by DOX. The specific NHE-1 inhibitor DMA can partly reverse the multidrug resistance of HL-60 cells induced by DOX.展开更多
Objective To test the resuscitative effects from prolonged ventricular fibrillation by epinephrine combined with sodium hydrogen exchanger isoform 1 inhibitor Cariporide. Methods 16 rats were received a 3 mg/kg bolu...Objective To test the resuscitative effects from prolonged ventricular fibrillation by epinephrine combined with sodium hydrogen exchanger isoform 1 inhibitor Cariporide. Methods 16 rats were received a 3 mg/kg bolus of Cariporide or the same volume of 0.9%NaCl solution (control) 15 seconds before completion 12 minutes untreated VF. Chest compression (CC) was started for a total of 8 minutes. Adjusted the depth of compressor so that the aortic diastolic pressure to 25～28 mmHg during the 2nd minute of CC. Fix the depth of the piston and this depth was used throughout the remaining 6 minutes of CC. 10 seconds before starting the 3rd minute of chest compression, injected epinephrine (30 μg/kg). Recorded the time at which restoration of spontaneous circulation (ROSC) occurred in Cariporide treated rats. Electrical defibrillation was timed in control group to match the time of spontaneous defibrillation in Cariporide treated rats. To the rats, which cant be defibrillated spontaneously, received chest compression and rescues electrical shocks. Results compared with control group, with the same CC depth, Cariporide treated rats received the higher and longer lasting coronary perfusion pressure (P< 0.05), higher resuscitative rate (P< 0.05), less post resuscitative ventricular ectopic activities (P< 0.001), better hemodynamic effects and longer survival time (P< 0.05). Conclusion Epinephrine combined with sodium hydrogen exchanger isoform 1 inhibitor Cariporide may represent a novel and remarkably effective intervention for resuscitation from prolonged VF.展开更多
文摘Objective: To explore the roles of intracellular pH value (pHi) and sodium-hydrogen exchanger isoform-1 (NHE-1) in the mechanism of multidrug resistance of leukemia cells. Methods: Multidrug resistant cell line HL-60 induced by doxorubicin(DOX) (called as HL-60/DOX cells) and their parent cell line HL-60 were employed as experiment group and control group. The proliferation and chemosensitivity of the cells were studied by MTT assay, and the expression of multidrug resistance protein (MRP) was detected by immol/Lunocytochemistry. Meanwhile, pHi was measured by spectrofluorometery with a fluorescence dye BCECF-AM. Based on the pHi recovery curve after intracellular acid loading, the activity of NHE-1 was analyzed. The expression of NHE-1 mRNA and MRP mRNA were determined by semi-quantitative RT-PCR. Cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and apoptotic DNA was extracted and electrophoresed. Results: ① The IC 50 values for DOX, MTZ, VCR and homoharringtonine(HT), in HL-60/DOX cells were significantly higher than those in HL-60 cells (P<0.01). HL-60/DOX cells expressed abundant MRP, but HL-60 cells did not. ② pHi of HL-60/DOX cells were significantly higher than that of HL-60 cells(P<0.001). The expression and activity of NHE-1 in HL-60/DOX cells were significantly stronger than those of HL-60 cells. ③After administration of the specific NHE-1 inhibitor dimethyl amiloride (DMA) at a certain range of concentrations, compared with HL-60 cells, the rate of growth inhibition of HL-60/DOX cells increased significantly (P<0.05), the drug-sensitivity of HL-60/DOX cells was significantly sensitive (P<0.01), the expression of MRP and MRP mRNA decreased significantly (P<0.01), the apoptosis rate increased significantly (P<0.01). Conclusion: NHE-1 is involved in the drug-resistant mechanisms of multidrug-resistant HL-60 cells induced by DOX. The specific NHE-1 inhibitor DMA can partly reverse the multidrug resistance of HL-60 cells induced by DOX.
文摘Objective To test the resuscitative effects from prolonged ventricular fibrillation by epinephrine combined with sodium hydrogen exchanger isoform 1 inhibitor Cariporide. Methods 16 rats were received a 3 mg/kg bolus of Cariporide or the same volume of 0.9%NaCl solution (control) 15 seconds before completion 12 minutes untreated VF. Chest compression (CC) was started for a total of 8 minutes. Adjusted the depth of compressor so that the aortic diastolic pressure to 25～28 mmHg during the 2nd minute of CC. Fix the depth of the piston and this depth was used throughout the remaining 6 minutes of CC. 10 seconds before starting the 3rd minute of chest compression, injected epinephrine (30 μg/kg). Recorded the time at which restoration of spontaneous circulation (ROSC) occurred in Cariporide treated rats. Electrical defibrillation was timed in control group to match the time of spontaneous defibrillation in Cariporide treated rats. To the rats, which cant be defibrillated spontaneously, received chest compression and rescues electrical shocks. Results compared with control group, with the same CC depth, Cariporide treated rats received the higher and longer lasting coronary perfusion pressure (P< 0.05), higher resuscitative rate (P< 0.05), less post resuscitative ventricular ectopic activities (P< 0.001), better hemodynamic effects and longer survival time (P< 0.05). Conclusion Epinephrine combined with sodium hydrogen exchanger isoform 1 inhibitor Cariporide may represent a novel and remarkably effective intervention for resuscitation from prolonged VF.
