新型katp开放剂埃他卡林对原代培养人肺动脉平滑肌细胞pcna mrna表达的影响
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江苏省自然科学基金资助(BK2006246)


effect of iptakalim,a novel katp opener,on the expression of proliferating cell nuclear antigen mrna in primary cultured human pulmonary artery smooth muscle cells
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    摘要:

    目的:观察新型atp敏感性钾通道(katp)开放剂埃他卡林(ipt)对原代培养人肺动脉平滑肌细胞增殖细胞核抗原(pcna)mrna表达的影响-方法:分离 3~4 级人肺小动脉,按照组织贴块法原代培养人肺动脉平滑肌细胞,分成对照组-缺氧组-缺氧+埃他卡林或吡那地尔组及缺氧+埃他卡林或吡那地尔+格列本脲组,应用实时荧光定量逆转录聚合酶链反应(fq-rt-pcr)技术检测细胞 pcna mrna表达-结果:缺氧使人肺动脉平滑肌细胞 pcna mrna表达增加(3.90±0.24)倍,与对照组比较差异有显著性(p < 0.01);相同条件下,在缺氧的同时,分别在培养液中加入 ipt 10-7-10-6-10-5 mol/l,pcna mrna表达分别下降(26.00±2.01)%-(55.33±4.04)%-(79.00±1.00)%,与缺氧组比较差异均有显著性(p均<0.01);加入 ipt 10-5 mol/l前 30 min,预先加入格列本脲(gli)10-5 mol/l,ipt的抑制作用抵消,与缺氧+ipt 10-5 mol/l组比较差异有显著性(p < 0.01)-结论:ipt通过激活细胞 katp通道,浓度依赖性地抑制缺氧时原代培养人肺动脉平滑肌细胞 pcna mrna表达的增加-

    Abstract:

    objective: to investigate the effect of iptakalim(ipt),a novel atp sensitive potassium(katp) opener,on the expression of proliferating cell nuclear antigen (pcna) mrna in primary cultured human pulmonary artery smooth muscle cells. methods:under germfree conditions,the smooth muscular layers were separated from human pulmonary arterioles(3rd~4th divisions). human pulmonary artery smooth muscle cells were cultured by tissue block method and were divided into following groups:control group,hypoxia group,hypoxia+iptakalim or pinacidil group and hypoxia+iptakalim or pinacidil+glibenclimide group. fluorescence quantitative reverse transcription-polymerase chain reaction(fq-rt-pcr)was used to measure the expression level of pcna mrna. results:the expression of pcna mrna in hypoxia group increased(3.90±0.24) times higher than that in control group. ipt at the concentrations of 10-7,10-6 and 10-5 mol/l lowered the expression of pcna mrna by (26.00±2.01)%,(55.33±4.04)%,(79.00±1.00)%,respectively,compared with that in hypoxia group(p < 0.01). glibenclimide (gli) at the concentration of 10-5 mol/l antagonized the effect of ipt with the same concentration on the expression of pcna mrna,compared with that in hypoxia + ipt 10-5 mol/l group(p < 0.01). conclusion:the expression of pcna mrna in human pulmonary artery smooth muscle cells under hypoxic conditions increased,while ipt has an inhibitory effect on the expression of pcna mrna in a concentration-dependent manner through activating katp channels.

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周燕娟,解卫平,张石江,李庆玲,朱煜明,王 虹.新型katp开放剂埃他卡林对原代培养人肺动脉平滑肌细胞pcna mrna表达的影响[J].南京医科大学学报(自然科学版),2011,(4):491-494

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  • 收稿日期:2011-09-10
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