摘要
目的:探讨新型内皮素受体拮抗剂GF-063和BQ-485对低氧培养的大鼠肺动脉平滑肌细胞(PASMCs)增殖的影响。方法:贴壁原代培养PASMCs。实验分为4组:常氧组(氧浓度210ml/L)、低氧组(氧浓度20ml/L)、低氧+内皮素A(ETA)受体拮抗剂BQ-485组(BQ-485的终浓度分别为1×10-6、1×10-7、1×10-8和1×10-9mo1/L)和低氧+GF-063组(GF-063的终浓度分别为1×10-6、1×10-7、1×10-8和1×10-9mo1/L)。上述4组细胞均分别培养24、48和72h。采用MTT比色法(波长492nm)检测BQ-485和GF-063对PASMCs增殖的影响(A值)。用流式细胞仪测定细胞周期;放射免疫检查法测定细胞培养上清液中内皮素-1(ET-1)的含量。结果:培养24h时,各实验组的A值差异不明显;培养48h时,低氧组的A值明显增加(P<0.01),低氧+BQ-485组和低氧+GF-063组在二者的终浓度为1×10-8、1×10-7、1×10-6mo1/L时A值下降,与低氧组比较具有统计学意义(P<0.01);在培养72h时,低氧组的A值仍高于常氧组,但较培养48h显著下降(P<0.05)。培养48h时,低氧组G2、S期细胞的比率及DNA合成增加,与常氧组比较有统计学意义(分别为P<0.01,P<0.05);与低氧组比较,低氧+BQ-485组和低氧+GF-063组G2、S期细胞的比率下降(P<0.05),G1期细胞增多(P<0.05),DNA合成减少。在培养48h时,与常氧组比,低氧组ET-1的水平增高(P<0.01),给予BQ-485、GF-063后ET-1的水平降低,BQ-485为1×10-7mol/L、GF-063为1×10-9mol/L时,可明显降低ET-1的水平(P<0.01)。结论:GF-063和BQ-485作为两种新型的内皮素受体拮抗都剂能抑制低氧培养的PASMCs增殖,减少ET-1的生成,且GF-063的其作用较BQ-485强。
AIM： To study the effects of a novel endothelin receptor antagonist GF-063 on the prolifera- tive behavior of cultured rat pulmonary arterial smooth muscle cells （PASMCs） by hypoxia stimulation. METHODS： Primary culture of rat PASMCs was prepared by tissue block anchorage. PASMCs were divided into four groups： normoxia （210 ml/L 02）, hypoxia （20 ml/L 02）, hypoxia ＋ BQ-485 （1×10^-6, 1×10^-7, 1 ×10^-8, 1×10^-9 mol/L） , and hypoxia ＋ GF-063 （1×10^-6, 1×10^-7, 1 ×10^-8, 1 ×10^-9 mol/L）. Proliferation of PASMCs was determined by MT-F （492 nm） assay and the cell cycle was measured by flow cytometry. Endothelin level in the supernatant was detected by radioimmunoassay. RESULTS： At 24 h, the A value exerted no significant effect on each group. At 48 h, the A value increased significantly in hypoxia group, but decreased significantly in both hypoxia ＋ BQ-485 group and hypoxia＋GF-063 （1 ×10^-6, 1 ×10^-7, 1 ×10^-8 mol/L） group, compared with hypoxia group （P〈 0.01, respectively）. At 72 h, the A value in the hypoxia group increased, but less than at 48 h （P 〈0. 05 ）. The percentage of cell and DNA synthesis of hypoxia increased significantly at G2 and S periods （P 〈0. 01, P 〈0. 05）. Compared with that in hypoxia group, the percentage of cell in hypoxia ＋ BQ- 485 group and hypoxia ＋ GF-063 group decreased at G2 and S periods （P 〈0. 05） , and the percentage of cell increased at G1 periods （P 〈 0.05 ）. The ET-1 level of supernatant in hypoxia group increased （P 〈 0.01）. The ET-1 level of supernatant in hypoxia group and BQ-485- （ 10-7 mol/L） treated groups and GF-063-（10-9 mol/L） treated groups decreased, respectively （P〈0.01）. CONCLUSION： GF-063 is a novel endothelin receptor antagonist that inhibits proliferation of PASMCs induced by hypoxia. GF-063, more than BQ-485, decreases the supernatant of ET-1 level. GF-063 is a promising drug in the treatment of puhnonary artery hypertension.
出处
《心脏杂志》
CAS
2010年第1期20-24,共5页
Chinese Heart Journal
基金
全军医药卫生科研课题基金项目资助(01MA049)
关键词
内皮素受体拮抗剂
低氧
肺动脉平滑肌细胞
增殖
endothelin receptor antagonist
cell hypoxia
pulmonary arterial smooth muscle cell
prolif- eration
