新型K_(ATP)开放剂埃他卡林对原代培养人肺动脉平滑肌细胞PCNAmRNA表达的影响  

Effect of iptakalim,a novel K_(ATP) opener,on the expression of proliferating cell nuclear antigen mRNA in primary cultured human pulmonary artery smooth muscle cells

作　　者：周燕娟[1,2] 解卫平[1] 张石江[3] 李庆玲[1] 朱煜明[1] 王虹[1] 

机构地区：[1]南京医科大学第一附属医院呼吸内科,江苏南京210029 [2]常州市武进人民医院呼吸内科,江苏常州213002 [3]南京医科大学第一附属医院胸外科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2011年第4期491-494,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省自然科学基金资助(BK2006246)

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 10-5mol/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。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.

关 键 词：ATP敏感性钾通道 埃他卡林 增殖细胞核抗原 人肺动脉平滑肌细胞 

分 类 号：R543.2[医药卫生—心血管疾病]