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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 observe the effect of iptakalim (IPT), a novel ATP sensitive potassium channel (KATP) opener, on the expression of proliferating cell nuclear antigen (PCNA) mRNA in primary cultured human pulmonary artery smooth muscle cells. Methods: Human pulmonary arterioles of grade 3 ~ 4 were isolated, and primary cultured human pulmonary artery smooth muscle cells were divided into control group, hypoxia group, hypoxia + iptakalim or pinacidil and hypoxia + , Eptakalim or pinacidil + glibenclamide group. The expression of PCNA mRNA was detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (FQ-RT-PCR). Results: Hypoxia increased the expression of PCNA mRNA in human pulmonary artery smooth muscle cells (3.90 ± 0.24) times, which was significantly different from the control group (P <0.01). Under the same conditions, hypoxia was added to the culture medium IPT 10-7, 10-6, 10-5 mol / L and PCNA mRNA decreased (26.00 ± 2.01)%, (55.33 ± 4.04)%, (79.00 ± 1.00)%, respectively, (P <0.01). Pretreatment with glibenclamide (GLI) 10-5 mol / L for 30 min before adding IPT 10-5mol / L, the inhibitory effect of IPT was offset by hypoxia + IPT 10-5 mol / L group, the difference was significant (P <0.01). CONCLUSION: IPT can inhibit the increase of PCNA mRNA expression in primary cultured human pulmonary artery smooth muscle cells in a concentration-dependent manner by activating KATP channel.