pp60c-src在高静水压培养的人脐静脉内皮细胞增殖中的作用

Role of pp60c-src in human umbilical vein endothelial cell proliferation induced by high hydrostatic pressure

目的 :探讨短期高压刺激是否诱导人脐静脉内皮细胞 (HUVECs)增殖以及pp60c -src所起的作用。方法 :分大气压 (AP ,0mmHg) ,中压 (MP ,12 0mmHg)和高压培养 (HP ,180mmHg) 3组 ,各组分别用卡托普利 (Cap ,10μmol/L)和厄贝沙坦 (Irb ,10 μmol/L)进行干预。测定 β -N -乙酰氨基己糖苷酶活性以反映细胞增殖。用Westernblotting法测定细胞内pp60c -src含量。结果 :① 2h时各组氨基己糖苷酶活性相似 ;中、高压组 4h时酶活性明显增高 (0 118± 0 0 0 3vs 0 14 5± 0 0 18和 0 14 4± 0 0 19,P <0 0 1) ,而大气压组 8h才见增高。 12h时各组酶活性相近。与此相应 ,2h时中、高压组pp60c -src明显高于大气压组 (0 12± 0 0 3vs 0 16± 0 0 2和 0 19± 0 0 6,均P <0 0 5) ,而大气压组 4h后才缓慢增加至中、高压组水平。② 12h时大气压组Cap与Irb对细胞数无影响 ,而在中、高压组两药使细胞增加 (MP组 0 189± 0 0 0 6vs 0 2 0 2± 0 0 0 5和 0 2 0 7± 0 0 0 3 ,P <0 0 5;HP组 0 193± 0 0 0 4vs 0 2 13± 0 0 0 4和0 2 2 1± 0 0 0 4,P <0 0 5)。但Cap与Irb干预组pp60c -src含量与对照组相似。结论 :压力刺激促使HUVECs早期 (4h)增殖 ,pp60c

AIM: To investigate whether or not short-term high pressur e proliferates human umbilical vein endothelial cells (HUVECs), and the role of pp60c-src in cell proliferation. METHODS: Cultured HUVECs of 3-6th passa ge were exposed to atmosphere 0 mmHg (AP), 120 mmHg (MP), 180 mmHg (HP) and interfered with cap top ril(Cap, 10 μmol/L)or irbesartan(Irb, 10 μmol/L) Cell proliferation was q uantified by measuring hexosaminidase activity. The content of pp 60 c-src in cells was investigated by Western blotting. RESULTS:① Hexos aminidase activities which reflected cell number increased significantly at 4 h in MP and HP(0 145±0 018 and 0 144±0 019 vs 0 118±0 003,P<0 01) Correspondingly, the expression of pp 60 c-src of 2 h in MP or HP gr oups was higher than that in AP whic h increased slow l y to reach the level in MP and HP groups at 4 h ② Cap and Irb had no effect o n c ell proliferation at each time points in AP group , but increased the hexosamin i dase activities in MP and HP groups at 12 h. The contents of pp 60 c-src in groups with or without intervention of Cap or Irb were similar at each time points. CON CLUSION: High hydrostatic pressure induces early cell proliferation. T he expre ssion of pp 60 c-src occurs before cell proliferation in different pressure groups , which suggests that pp 60 c-src is involved in signal transmission pathwa y of pr essure-induced cell proliferation. This process is not regulated mainly by renin -angiotensin system (RAS).