目的:观察胍基丁胺对低氧培养大鼠肺动脉平滑肌细胞(PASMCs)增殖的影响。方法:采用组织块贴壁法原代培养大鼠PASMCs,取对数生长期PASMCs分对照组和胍基丁胺组,比色法测定细胞培养液中乳酸脱氢酶(LDH)活力,液闪计数仪测定[^3H]-TdR掺入量,流式细胞仪测定细胞周期,图像分析法测定增殖细胞核抗原含量(PCNA)作为细胞增殖的指标。结果:常氧和低氧(2.5%O2)培养时,胍基丁胺组LDH活力与对照组LDH活力均无显著差异(P〉0.05);与单纯低氧组相比,50μmol/L胍基丁胺可显著减少低氧培养PASMCs的[^3H]-TdR掺入量(P〈0.01)。随着胍基丁胺浓度从50μmol/L逐渐增加至500μmol/L,大鼠PASMCs[^3H]-TdR的掺入量亦随之减少(P〈0.01)。与单纯低氧组相比,胍基丁胺可显著减少低氧培养PASMCs的PCNA的含量(P〈0.01),使G0/G1期细胞比例显著增加,G2/M期细胞比例显著减少(P〈0.01)。结论:胍基丁胺对PASMCs不产生明显的细胞毒性作用;胍基丁胺可抑制低氧培养大鼠PASMCs的增殖,这种抑制作用呈剂量依赖性。
AIM: To determine the effect of agmatine on the proliferation of pulmonary artery smooth muscle cells (PASMCs) induced by hypoxia. METHODS: Primary culture of rat PASMCs was prepared from adult male Wistar rat pulmonary artery by the method of tissue block anchorage. PASMCs were divided into two groups: control group and agmatine- treated group. The activity of LDH in the medium was measured by chromatometry. [^3H] - TdR incorporation was measured by liquid scintillation counting. The cell cycle was measured by flow cytometry, and the PCNA content was measured by image analysis.RESULTS: Agmarine did not exert significant effect on the activity of LDH in the medium, but significantly decreased the [^3H] - TdR incorporation and PCNA content of PASMCs. Agmatine significantly decreased the ceil ratio of G2/M phase and increased the cell ratio of G0/G1 phase. With the increment of the concentration of agmatine, [^3H] - TdR incorporation was significantly decreased. CONCLUSION: Agmatine has no significant cytotoxic effect on PASMCs and agmatine dose- dependently inhibits the proliferation of rat PASMCs induced by hypoxia.