阿伐他汀对幼年大鼠心肌成纤维细胞增殖和胶原合成的影响及其意义

目的　探讨阿伐他汀对幼年大鼠心肌成纤维细胞 (CF)增殖和胶原合成的影响。方法　用消化法培养新生SD大鼠的CF ,采用3H 胸腺嘧啶核苷 ( 3H TdR)掺入法测定CF的DNA合成功能 ,四氮唑蓝 (MTT)比色法测定细胞增殖 ,流式细胞分析仪技术检测细胞周期 ,3H 脯氨酸掺入法测定胶原合成 ,分别观察不同浓度阿伐他汀对CF增殖和胶原合成的影响。结果　 ( 1)阿伐他汀以浓度依赖的方式抑制CF的3H TdR掺入率。其中 ,10 -7、10 -6、10 -5和 10 -4 mol/L组的3H TdR掺入率分别为(cpm/ 5 0 0 0细胞 ) 314± 6 8、2 5 3± 5 2、2 0 1± 5 3和 170± 48,显著低于对照组 ( 378± 6 5 ) (F =16 912 ,P <0 0 0 1)。 ( 2 )随着阿伐他汀浓度的增高 ,CF的MTT比色法A4 90 值呈明显的递减趋势 ,其中 10 -7、10 -6、10 -5和 10 -4 mol/L组的A4 90 值分别为 0 2 88± 0 0 0 8、0 2 5 2± 0 0 0 7、0 2 2 5± 0 0 0 8和 0 2 16± 0 0 13,与对照组 ( 0 311± 0 0 0 5 )相比 ,差异有非常显著意义 (P均 <0 0 1)。 ( 3)G0 /G1期细胞百分率随阿伐他汀浓度的增高而呈递增趋势 ,S期G2 /M期细胞百分率和增殖指数呈递减趋势 ,其中 10 -7、10 -6、10 -5和10 -4 mol/L组与对照组比较 ,差异有非常显著意义 (P均 <0 0 1)。 ( 4)CF的3H 脯

Effects of atorvastatin on the proliferation and collagen synthesis of rat cardiac fibroblasts

OBJECTIVE: To investigate the effects of atorvastatin on the proliferation and collagen synthesis of rat cardiac fibroblasts (CFs). METHODS: Isolated and cultured CFs of neonatal Sprague-Dawley (SD) rats were isolated and cultured. The DNA synthesis of CFs was measured by (3)H-TdR uptake test. CFs proliferation was measured by thiazolyl blue (MTT) assay. Cell cycle distribution was determined with flow cytometer (FCM). Collagen synthesis was measured by 3H-proline uptake test. RESULTS: (1) The (3)H-TdR uptake in the 10(-7) mol/L, 10(-6) mol/L, 10(-5) mol/L, and 10(-4) mol/L atorvastatin groups was (cpm/5,000 cells) 314 +/- 68, 253 +/- 52, 201 +/- 53, and 170 +/- 48 respectively, all significantly lower than that of the control group (378 +/- 65, all P < 0.001). (2) MTT colorimetry showed that the A(490) values were 0.288 +/- 0.008, 0.252 +/- 0.007, 0.225 +/- 0.008, and 0.216 +/- 0.013 respectively in the 10(-7) mol/L, 10(-6) mol/L, 10(-5) mol/L and 10(-4) mol/L atorvastatin groups, all significantly lower than that in the control group (0.311 +/- 0.005, all P < 0.01). (3) The percentage of cells in G(0)/G(1) phase was 54.8% +/- 2.5%, 61.4% +/- 2.7%, 70.4% +/- 3.2%, and 82.0% +/- 4.0% in the 10(-7) - 10(-4) mol/L atorvastatin groups, all significantly higher than that in the control group (46.5 +/- 2.9, all P < 0.01). The percentage of cells in S phase was 18.8% +/- 2.3%, 15.8% +/- 2.1%, 12.5% +/- 1.8%, and 7.3% +/- 2.0% in the 10(-7) - 10(-4) mol/L atorvastatin groups, all significantly lower than that in the control group (23.1% +/- 2.0%, all P < 0.01). The percentage of cells in G(2)/M phase was 26.5% +/- 0.8%, 22.8% +/- 1.2%, 17.2% +/- 1.4%, and 10.7% +/- 2.0% in the 10(-7) - 10(-4) mol/L atorvastatin groups respectively, all significantly lower than that in the control group (30.5% +/- 1.4%, all P < 0.01). The proliferation index (PI) was 45.3% +/- 2.5%, 38.6% +/- 2.7%, 29.6% +/- 3.2%, and 18.0% +/- 4.0% in the 10(-7) - 10(-4) mol/L atorvastatin groups respectively, all significantly lower than that in the control group (53.5% +/- 2.9%, all P < 0.01). (4) The (3)H-proline uptake was (cpm/5 000 cells) 422 +/- 59, 332 +/- 67, 252 +/- 53, and 184 +/- 48 in the 10(-7) mol/L, 10(-6) mol/L, 10(-5) mol/L, and 10(-4) mol/L atorvastatin groups respectively, all significantly lower than that in the control group (566 +/- 62, all P < 0.01). CONCLUSION: Atorvastatin inhibits effectively the proliferation of cultured rat CFs and the collagen synthesis therein, which may play a role in the regression of heart remodeling.