一氧化氮及一氧化碳对碱性成纤维细胞生长因子促血管平滑肌细胞增殖作用的影响

Effect of nitric monoxide and carbon monoxide on bFGF stimulated proliferation of vascular smooth muscle cells

目的：研究一氧化氮（ＮＯ） 及一氧化碳（ＣＯ） 对碱性成纤维细胞生长因子（ｂａｓｉｃ ｆｉｂｒｏｂｌａｓｔ ｇｒｏｗｔｈ ｆａｃｔｏｒ，ｂＦＧＦ） 促平滑肌细胞（ｖａｓｃｕｌａｒｓｍｏｏｔｈ ｍｕｓｃｌｅ ｃｅｌｌｓ， ＶＳＭＣ）增殖作用的影响及其机制。方法：以３ＨＴｄＲ参入法测定平滑肌细胞增殖程度，放射活性法测定ＶＳＭＣ内蛋白磷酸化程度、蛋白激酶Ｃ（ｐｒｏｔｅｉｎ ｋｉｎａｓｅ Ｃ，ＰＫＣ） 及丝裂素活化蛋白激酶（ｍｉｔｏｇｅｎａｃｔｉｖａｔｅｄ ｐｒｏｔｅｉｎ ｋｉｎａｓｅ， ＭＡＰＫ）活性。结果：孵育２４ ｈ 后，ｂＦＧＦ刺激的ＶＳＭＣ增殖较对照组增加１．１ 倍（Ｐ＜ ０．０１），细胞内蛋白磷酸化程度增加３８．２％ （Ｐ＜ ０．０１），ＰＫＣ 及ＭＡＰＫ 活性分别增加１．５ 和２ ．５倍（ Ｐ＜ ０．０１）；ＣＯ 前体ｈｅｍｅＬｌｙｓｉｎａｔｅ（ＨＬＬｙｓ） 及ＮＯ 前体ＬＡｒｇｉｎｉｎｅ（ ＬＡｒｇ） 对静止期ＶＳＭＣ 生长无显著影响，亦未引起细胞内蛋白磷酸化、ＰＫＣ及ＭＡＰＫ 活性发生显著改变；５、１０ 、２０ μｍｏｌ·Ｌ－１ ＨＬＬｙｓ 使ｂＦＧＦ刺激的ＶＳＭＣ增殖分别减少４３．４ ％ 、４６．６％ 和４７ ．６％

Objective: To investigate the effect of nitric oxide(NO) and carbon monoxide(CO) on proliferation of vascular smooth muscle cells(VSMC) stimulated by basic fibroblast growth factor(bFGF) and their mechanism. Methods: VSMC proliferation was measured by 3H TdR incorporation. Protein phosphorylation, PKC and MAPK activity in VSMC were determined by radioactivity assay. Results: Compared with control, VSMC amplification induced by bFGF was increased by 1.1 fold after 24 h incubation ( P <0.01). Protein phophorylation, PKC and MAPK activities in VSMC were augmented by 38.2%, 1.5 and 2.5 times, respectively ( P <0.01); H L Lys and L Arg had no significant influence on both growth and intracellular protein phosphorylation, PKC and MAPK activities in quiescent VSMC. 5, 10 and 20 μmol·L -1 H L Lys decreased bFGF stimulated VSMC proliferation by 43.4%,46.6% and47.6%,respectively ( P <0.01). Intracellular PKC activities were lowered by 25.6%, 31.8% and 33.9%, respectively ( P <0.01), and MAPK activities were reduced by 15.0%, 25.5% and 40.1%, respectively ( P <0.01); 5, 10 and 20 mmol·L -1 L Arg decreased bFGF stimulated VSMC proliferation by 30.2%, 40.5% and 51.9%, respectively ( P <0.01). Intracellular PKC activities were lowered by 29.8%, 28.9% and 36.6%,respectively ( P <0.01), and MAPK activities were reduced by 19.4%, 32.4% and 44.4%, respectively ( P <0.01). bFGF induced protein phosphorylation in VSMC was inhibited by 10 μmol·L -1 H L Lys and 10 mmol·L -1 L Arg by 31.8% and 37.4%, respectively ( P <0.01). Conclusion: NO and CO may have an inhibitive role against VSMC proliferation caused by bFGF, whose mechanism has a close relationship with inhibition of intracellular protein phosphorylation, PKC and MAPK activities stimulated by bFGF.