Objective：To observe the protective effects of erythropoietin （EPO） pretreatment on cardiac myocyte with hypoxia/reoxygenation （H/R） injury and the role of NF-κBin this effects. Methods：After the H/R model of c...Objective：To observe the protective effects of erythropoietin （EPO） pretreatment on cardiac myocyte with hypoxia/reoxygenation （H/R） injury and the role of NF-κBin this effects. Methods：After the H/R model of cardiac myocytes of neonatal rats was established, the cultured cardiac myocytes were divided into 4 groups, including EPO pretreatment group （ EPO 10 U/ml 24 h before H/R）, EPO pretreatment ＋ PDTC group（EPO 10 U/ml and PDTC 5 μg/ml 24 h before H/R）, PDTC group （PDTC 5 μg /ml 24 h before H/R） and eomrolgroup. Before and after the H/R, assay of LDH concentration in the culture medium, the survival rate of the myocytes tested by MTT chromatometry and the apoptosis by flow cytometry were undertaken. Activation of NF-κB was determined by EMSA before and after H/R. Results：EPO pretreatment markedly reduced the LDH concentration in the medium, elevated the survival rate of myocytes and inhibited the apoptosis after H/R. Addition of PDTC during the pretreatment abol- ished the protective effects of EPO pretreatment. NF-κB was markedly activated during EPO pretreatment and PDTCinhibited the activation. However, after H/R, the activity of NF-κB in myocytes with EPO pretreatment was significantly inhibited compared to the other myocytes. Conclusion：NF-κB is significantly activated during EPO pretreatment, but is inhibited after H/R, which is correlated with the protective effects of EPO pretreatment on cardiac myocytes with H/R. This phenomenon can be explained as the negative feedback mechanism of the activation of NF-κB.展开更多
Objective: To study the effect of KATp channel opener and its possible mechanism on the sinoatrial node cells of neonatal rats which were cultured under simulated ischemia-reperfusion. Methods: Freshly isolated sinoat...Objective: To study the effect of KATp channel opener and its possible mechanism on the sinoatrial node cells of neonatal rats which were cultured under simulated ischemia-reperfusion. Methods: Freshly isolated sinoatrial node (SAN) cells of neonatal rats were purified and cultured for 2 d, and then they were randomly divided into the control, simulated ischemia-reperfusion group (I/R group) , group intervened with KATp channel opener pinacidil (P + I/R group), KATP Channel blocking agent 5-HD (5-HD + I/R group) , and group with the 2 agents at same time (5-HD + P + I/R group) . The survival rate of cells was measured by flow cytometry and the content of intracellular calcium in the cells of each group was detected with laser confocal microscopy. Results: ① The survival rate of SAN cells in I/R group [ (51. 79±6. 28)% ] was remarkably significantly lower than in control [ (95. 08±10. 48)% ] (P < 0.001), and very significantly lower than in P + I/R group [ (63. 77±5. 35) % ] (P<0.01), however, those of 5-HD + P + I/R group [(52. 88±6. 25)% ] and 5-HD+I/R group [ (53. 16±5. 35)% ] was significantly lower compared with that in P + I/R group (P <0. 01) ; ② When the average fluorescence intensity of sinoatrial node cells in the control was regarded as 100% , the relative fluorescence intensities of each group were: ( 374±52) % in I/R group, significantly higher than that of control (P <0. 01) ; ( 162±20)% in P + I/R group, declining significantly than that of I/R group (P<0.01); (385?6)% in 5-HD+ P + I/R group and (379±44)% in 5-HD + I/R group, increasing significantly than that of I/R group (P<0.01). Conclusion: ① Simulated ischemia-reperfusion can significantly reduce the survival rate of SAN cells, and can also lead to the overload of intracellular calcium in them.② KATp channel opener, pinacidil, exerts protective effect on the cells under simulated ischemia-reperfusion, which may be associated with the decrease of intracellular calcium loading in them.展开更多
文摘Objective：To observe the protective effects of erythropoietin （EPO） pretreatment on cardiac myocyte with hypoxia/reoxygenation （H/R） injury and the role of NF-κBin this effects. Methods：After the H/R model of cardiac myocytes of neonatal rats was established, the cultured cardiac myocytes were divided into 4 groups, including EPO pretreatment group （ EPO 10 U/ml 24 h before H/R）, EPO pretreatment ＋ PDTC group（EPO 10 U/ml and PDTC 5 μg/ml 24 h before H/R）, PDTC group （PDTC 5 μg /ml 24 h before H/R） and eomrolgroup. Before and after the H/R, assay of LDH concentration in the culture medium, the survival rate of the myocytes tested by MTT chromatometry and the apoptosis by flow cytometry were undertaken. Activation of NF-κB was determined by EMSA before and after H/R. Results：EPO pretreatment markedly reduced the LDH concentration in the medium, elevated the survival rate of myocytes and inhibited the apoptosis after H/R. Addition of PDTC during the pretreatment abol- ished the protective effects of EPO pretreatment. NF-κB was markedly activated during EPO pretreatment and PDTCinhibited the activation. However, after H/R, the activity of NF-κB in myocytes with EPO pretreatment was significantly inhibited compared to the other myocytes. Conclusion：NF-κB is significantly activated during EPO pretreatment, but is inhibited after H/R, which is correlated with the protective effects of EPO pretreatment on cardiac myocytes with H/R. This phenomenon can be explained as the negative feedback mechanism of the activation of NF-κB.
基金Supported by National Natural Science Foundation of China (No. 30070314)
文摘Objective: To study the effect of KATp channel opener and its possible mechanism on the sinoatrial node cells of neonatal rats which were cultured under simulated ischemia-reperfusion. Methods: Freshly isolated sinoatrial node (SAN) cells of neonatal rats were purified and cultured for 2 d, and then they were randomly divided into the control, simulated ischemia-reperfusion group (I/R group) , group intervened with KATp channel opener pinacidil (P + I/R group), KATP Channel blocking agent 5-HD (5-HD + I/R group) , and group with the 2 agents at same time (5-HD + P + I/R group) . The survival rate of cells was measured by flow cytometry and the content of intracellular calcium in the cells of each group was detected with laser confocal microscopy. Results: ① The survival rate of SAN cells in I/R group [ (51. 79±6. 28)% ] was remarkably significantly lower than in control [ (95. 08±10. 48)% ] (P < 0.001), and very significantly lower than in P + I/R group [ (63. 77±5. 35) % ] (P<0.01), however, those of 5-HD + P + I/R group [(52. 88±6. 25)% ] and 5-HD+I/R group [ (53. 16±5. 35)% ] was significantly lower compared with that in P + I/R group (P <0. 01) ; ② When the average fluorescence intensity of sinoatrial node cells in the control was regarded as 100% , the relative fluorescence intensities of each group were: ( 374±52) % in I/R group, significantly higher than that of control (P <0. 01) ; ( 162±20)% in P + I/R group, declining significantly than that of I/R group (P<0.01); (385?6)% in 5-HD+ P + I/R group and (379±44)% in 5-HD + I/R group, increasing significantly than that of I/R group (P<0.01). Conclusion: ① Simulated ischemia-reperfusion can significantly reduce the survival rate of SAN cells, and can also lead to the overload of intracellular calcium in them.② KATp channel opener, pinacidil, exerts protective effect on the cells under simulated ischemia-reperfusion, which may be associated with the decrease of intracellular calcium loading in them.
