Jin Hong, Lin Jijin, Fu Lu, Mei Yi-Fang, Peng Geng, Tan Xuerui, Wang Dong-Ming, Wang Wei, Li Yu-Guang
Department of Cardiology, The First Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong 515041, China.
Biochem Cell Biol. 2007 Apr;85(2):246-51. doi: 10.1139/O07-011.
There is a striking gender difference in atherosclerotic vascular disease. For decades, testosterone was considered detrimental to the cardiovascular system. Recent studies, however, have presented some alternative results. The aim of this study was to evaluate the effect of testosterone, using physiological and supraphysiological concentrations, on antigen and mRNA levels of tissue plasminogen activator (tPA), plasminogen activator inhibitor type 1 (PAI-1), and tissue factor pathway inhibitor (TFPI) released by human umbilical vein endothelial cells and to investigate the cellular mechanism. Cells within 2-3 passages were cultured in 25 cm(2) flasks or plated onto 96-well plates with a density of about 1 x 10(5) cells/mL as recommended. The cells were incubated in the presence or absence of testosterone (3, 30, 3 x 10(3), 3 x 10(4) nmol/L) for 48 h. Levels of tPA, PAI-1, and TFPI antigen were assayed with ELISA kits. Reverse transcriptase PCR was carried out to detect tPA, PAI-1, and TFPI mRNA levels. Cells were incubated in androgen-receptor antagonist (flutamide 10 micromol/L) or aromatase inhibitor (aminoglutethimide 50 micromol/L) for 3 h, and then the experiments were repeated. Testosterone at a physiologic concentration (30 nmol/L) increased the antigen levels of tPA and TFPI significantly (P < 0.05). However, tPA and TFPI levels were markedly reduced (P < 0.05) at a larger dose (3 x 10(4) nmol/L). On the other hand, PAI-1 antigen levels decreased significantly at the testosterone concentrations ranging from 3 to 3 x 10(4) nmol/L (P < 0.05). The change in the levels of tPA and TFPI were reflected in the corresponding change in mRNA levels. Flutamide attenuated the effect of testosterone at physiological concentration (30 nmol/L). The results demonstrated that testosterone at physiological concentrations may have a beneficial influence on the haemostatic system through enhancement of anticoagulant activity, resulting from stimulation of TFPI and tPA expression and inhibition of PAI-1 secretion by the endothelium.
动脉粥样硬化性血管疾病存在显著的性别差异。数十年来，睾酮被认为对心血管系统有害。然而，最近的研究呈现了一些不同的结果。本研究的目的是评估生理浓度和超生理浓度的睾酮对人脐静脉内皮细胞释放的组织型纤溶酶原激活物（tPA）、纤溶酶原激活物抑制剂1型（PAI-1）和组织因子途径抑制剂（TFPI）的抗原及mRNA水平的影响，并探究其细胞机制。按照推荐，将传代2至3次的细胞接种于25 cm²培养瓶中，或以约1×10⁵个细胞/mL的密度接种到96孔板上。细胞在有或无睾酮（浓度为3、30、3×10³、3×10⁴ nmol/L）的条件下孵育48小时。使用ELISA试剂盒检测tPA、PAI-1和TFPI抗原水平。进行逆转录聚合酶链反应以检测tPA、PAI-1和TFPI的mRNA水平。细胞在雄激素受体拮抗剂（氟他胺10 μmol/L）或芳香化酶抑制剂（氨鲁米特50 μmol/L）中孵育3小时，然后重复实验。生理浓度（30 nmol/L）的睾酮显著提高了tPA和TFPI的抗原水平（P < 0.05）。然而，在更高剂量（3×10⁴ nmol/L）时，tPA和TFPI水平显著降低（P < 0.05）。另一方面，在睾酮浓度为3至3×10⁴ nmol/L范围内，PAI-1抗原水平显著降低（P < 0.05）。tPA和TFPI水平的变化反映在相应的mRNA水平变化上。氟他胺减弱了生理浓度（30 nmol/L）睾酮的作用。结果表明，生理浓度的睾酮可能通过增强抗凝活性对止血系统产生有益影响，这是由于刺激了TFPI和tPA的表达以及抑制了内皮细胞分泌PAI-1所致。
