碱性成纤维细胞生长因子反义寡脱氧核苷酸抑制胰岛素诱导动脉平滑肌细胞增殖

Basic Fibroblast Growth Factor Antisense Oligodeoxynucleotides Inhibited Proliferation of Smooth Muscle Cells Induced by Insulin

为研究胰岛素诱导的动脉平滑肌细胞增殖作用和碱性成纤维细胞生长因子反义寡脱氧核苷对培养的Ｗｉｓｔａｒ大鼠主动脉平滑肌细胞生长的影响。采用胰岛素和碱性成纤维细胞生长因子反义寡脱氧核苷酸处理培养的Ｗｉｓｔａｒ大鼠主动脉平滑肌细胞，Ｎｏｒｔｈｅｎｂｌｏｔ检测碱性成纤维细胞生长因子基因ｍＲＮＡ表达，并测定氚标胸腺嘧啶脱氧核苷掺入和细胞计数。结果发现胰岛素能明显诱导平滑肌细胞碱性成纤维细胞生长因子ｍＲＮＡ表达和增殖，且呈浓度依赖性。胰岛素浓度由０．００１ｍｇ／Ｌ增加到１．０ｍｇ／Ｌ，平滑肌细胞增殖率由１０％增加到５３％。碱性成纤维细胞生长因子反义寡脱氧核苷酸（５μｍｏｌ／Ｌ）能明显抑制胰岛素诱导的平滑肌细胞碱性成纤维细胞生长因子ｍＲＮＡ表达和增殖，氚标胸腺嘧啶脱氧核苷掺入被抑制４１．１％（与对照组比较，Ｐ＜０．０１），细胞计数被抑制３０．２％（与对照组比较，Ｐ＜０．０１）。提示胰岛素可通过诱导平滑肌细胞碱性成纤维细胞生长因子的表达而促进动脉平滑肌细胞增殖，碱性成纤维细胞生长因子反义寡脱氧核苷酸能有效抑制胰岛素诱导的平滑肌细胞的ＤＮＡ合成及其增殖。

Aim To study effect of insulin and basic fibroblast growth factor (bFGF) antisense oligodeoxynucleotides (ODN) on the growth of cultured rat artery smooth muscle cells (SMCs). Methods Antisense and sense ODNs of bFGF were added into SMC medium, bFGF gene expression was detected by Northern bloting, cell hyperplasia was evaluated by thymidine incorporation and cell counting. Results Insulin induced proliferation of cultured vascular SMCs in concentration-dependent way and bFGF mRNA expression. Insulin increased from 0.001 mg/L to 1.0 mg/L, the rate of SMC hyperplasia from 10% to 53%. Antisense ODN (5 μmol/L) of bFGF significantly inhibited proliferation of SMCs, thymidine incorporation by 41.1% (P<0.01), and cell number by 30.2% (P<0.01 vs control) respectively. Conclusion Insulin may enhance SMC proliferation by inducing bFGF gene expression, antisense ONDs of bFGF inhibited partly DNA synthesis and proliferation of cultured vascular SMC induced by insulin.