辛伐他汀对鼠心脏成纤维细胞DNA合成的影响

目的探讨辛伐他汀(Sim)对新生SD大鼠心脏成纤维细胞(CFs)DNA合成的影响及甲羟戊酸(MVA)的干预效应。方法采用胰酶消化、差速贴壁法培养新生SD大鼠CFs,以3H-胸腺嘧啶核苷(3H-TdR)掺入法测定DNA合成,观察不同浓度Sim及MVA分别作用不同时间对CFsDNA合成功能的影响。结果 (1)CFs的3H-TdR掺入率随着Sim干预浓度的增加而降低,其中10-6和10-5mol／L Sim组的3H-TdR掺入率分别为(1175±202．66)、(771±164．86)cpm／2000细胞, 显著低于对照组的(1608±204．32)cpm／2000细胞(均P<0．01);(2)10-5mol／L Sim作用CFs 6、12、18、24、30、36、42、48 h, 随着Sim作用时间的延长,CFs的3H-TdR掺入率呈递减趋势,与时间呈明显的负相关(r=-919,P<0．01);(3)CFs的3H-TdR 掺入率随着MVA干预浓度的增加呈进行性上升,其中10-4、10-3 mol／L MVA 10-5 mol／LSim组3H-TdR掺入率分别为 (1612±308．57)、(1995±353．83)cpm／2000细胞,显著高于对照10-5mol/L Sim组的(771±164．86)cpm／2000细胞(P<0．01); (4)10-3 mol／L MVA分别作用CFs 6-48 h,CFs的3H-TdR掺入率随着MVA作用时间的延长而逐渐增加,呈明显的正相关(r=0．968,P<0．01)。结论 Sim呈浓度-时间依赖方式抑制CFs DNA的合成且可被MVA拮抗,提示Sim可抑制CFs 增殖,延缓心肌纤维化,其作用可能通过MVA途径实现。

Effects of simvastatin on DNA synthesis in rat cardiac fibroblasts

OBJECTIVE: To investigate the effects of simvastatin (Sim) and the interference by mevalonate (MVA) against its effect on DNA synthesis in rat cardiac fibroblasts (CFs). METHODS: CFs were isolated from neonatal SD rats by trypsin digestion and growth-arrested CFs were stimulated with Sim and/or MVA at varied concentrations for different time lengths, and the DNA synthesis in the cells was measured by (3)H-thymidine ((3)H-TdR) incorporation assay. RESULTS: Sim decreased (3)H-TdR incorporation in the CFs in a concentration-dependent manner, and (3)H-TdR incorporation was significantly lower in cells treated with 1 x 10(-6) and 1 x 10(-5) mol/L Sim (1,175+/-202.66 and 771+/-164.86 cpm/2000 cells, respectively) than in the control cells (1,608+/-204.32 cpm/2000 cells, P<0.01). As the treatment time with 1 x 10(-5) mol/L Sim prolonged (for 6, 12, 18, 24, 36, 42, and 48 h), (3)H-TdR incorporation in CFs decreased gradually, showing an obvious inverse correlation with the treatment time (r=-919, P<0.01). (3)H-TdR incorporation in cells treated with 1 x 10(-6) to 1 x 10(-3) mol/L MVA and 1 x 10(-5) mol/L Sim rose steadily as MVA concentration increased. A significant difference in the incorporation was found between cells treated with both 1 x 10(-4)/1 x 10(-3) mol/L MVA and 1 x 10(-5) mol/L Sim (1,612+/-308.57 and 1,995+/-353.83 cpm/2000 cells, respectively) and the cells with 1 x 10(-5) mol/L Sim treatment alone (P<0.01); difference was also noted between cells treated with 1 x 10(-5) mol/L MVA and the control cells (P<0.05), but treatment with 1 x 10(-6) mol/L MVA did not produce much difference in comparison with the control cells (P>0.05) With the increase of treatment time (for 6, 12, 18, 24, 36, 42, 48 h), 1 x 10(-3) mol/L MVA caused steady increase in (3)H-TdR incorporation in the CFs, showing a significant positive correlation with the treatment time (r=0.968, P<0.01). CONCLUSION: Sim can decrease DNA synthesis in rat CFs and postpone the occurrence of myocardial fibrosis, which can be reversed by MVA.