溶血磷脂酰胆碱刺激脑微血管平滑肌细胞增殖的细胞内信号转导途径

Signal transduction pathways stimulated by lysophosphatidylcholine in cultured bovine cerebral microvascular smooth muscle cells

通过测定 [3 H]胸腺嘧啶核苷 ( [3 H]Td R)参入和结晶紫染色法测定平滑肌细胞增殖 ,研究了溶血磷脂酰胆碱 ( LPC)刺激牛脑微血管平滑肌细胞( BCMSMC)增殖的细胞内信号转导途径 .结果显示 ,LPC能浓度依赖性 ( 1 nmol· L-1- 1 0μmol·L-1)诱导 BCMSMC摄取 [3 H]Td R,在 LPC的浓度为 1 0μmol· L-1时作用达最大 ,cpm由 366± 1 42升至 1 761± 2 96( P<0 .0 1 ) ;LPC亦能浓度依赖性( 1 nmol· L-1- 1 0μmol· L-1)诱导 BCMSMC增殖 ,在 LPC浓度为 1 μmol· L-1时促增殖作用达坪值 ,A595nm由 0 .0 60± 0 .0 0 9增至 0 .1 0 0± 0 .0 1 5( P<0 .0 1 ) .丝裂原激活蛋白激酶 ( MAPK)特异性抑制剂 PD980 59( 2 - 50 μmol· L-1) ,血小板衍生生长因子受体抑制剂酪氨酸磷酸化抑制剂 AG 1 2 96( 2 -50 μmol· L-1)以及蛋白质酪氨酸激酶抑制剂除莠霉素 A( 2 - 1 0μmol· L-1)能浓度依赖性地抑制LPC的上述作用 .表明 LPC能促进 BCMSMC增殖 ,其细胞内信号转导与 MAPK途径有关 .

The signal transduction pathways stimulated by lysophosphatidylcholine (LPC) in bovine cerebral microvascular smooth muscle cells (BCMSMC) were studied. The [ 3H]TdR incorporation and crystal violet assay were measured in BCMSMC. The results showed that LPC enhanced the BCMSMC [ 3H]TdR incorporation in a concentration (1 nmol·L -1 -10 μmol·L -1 ) dependent manner. At 10 μmol·L -1 , the incorporation rate reached maximum (cpm from 366±142 to 1761±296, P<0.01); LPC also promoted the proliferation of BCMSMC in a concentration (1 nmol·L -1 -10 μmol·L -1 ) dependent manner. At 1 μmol·L -1 , the effect reached plateau level (A 595 nm from 0.060±0.009 to 0.100±0.015). Mitogen activated protein kinase (MAPK) inhibitor PD 98059 (2-50 μmol·L -1 ), platelet derived growth factor receptor inhibitor tyrphostin AG 1296 (2-50 μmol·L -1 ) and protein tyrosine kinase inhibitor herbimycin A (2-10 μmol·L -1 ) inhibited the BCMSMC [ 3H]TdR incorporation and proliferation of BCMSMC. The data indicate that LPC may stimulate the proliferation of BCMSMC. The LPC induced intracellular signal transduction may be related to the MAPK pathway.