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目的观察拉西地平对结缔组织生长因子(CTGF)诱导大鼠心肌细胞肥大的作用,探讨其作用与细胞外信号调节激酶1/2(ERK1/2)的关系。方法以培养的新生SD大鼠的心肌细胞为实验模型,用图象分析法、3H-亮氨酸掺入法、考马斯亮蓝染色、蛋白免疫印迹法(Westernblot)等,评价拉西地平对CTGF诱导心肌细胞肥大的抑制作用及其与ERK1/2的关系。结果①以50ng/L的CTGF作用24h后,心肌细胞表面积为(1812.52±168.73)μm2,与对照组(689.31±96.58)μm2比较显著增加(P<0.01);5、10、25及50μmol/L拉西地平干预组的心肌细胞表面积分别为(1476.52±156.73)μm2、(1120.39±149.68)μm2、(926.10±101.44)μm2及(739.81±91.55)μm2,与CTGF组比较呈浓度依赖性降低(P<0.01)。②以50ng/L的CTGF作用24h后,心肌细胞的3H-亮氨酸掺入率为(2368.72±122.45)cpm/孔,与对照组(950.26±89.43)cpm/孔比较显著增加(P<0.01)。5、10、25及50μmol/L拉西地平干预组的心肌细胞的3H-亮氨酸掺入率,分别为(2023.12±106.15)cpm/孔、(1629.15±103.46)cpm/孔、(1302.19±98.53)cpm/孔及(1055.72±90.96)cpm/孔,与CTGF组比较呈浓度依赖性降低(P<0.01)。③以50ng/L的CTGF作用24h后,心肌细胞蛋白的含量为(1.692±0.203)ng/细胞,与对照组(0.622±0.068)ng/细胞比较显著增加(P<0.01);5、10、25及50μmol/L拉西地平干预组的心肌细胞蛋白的含量分别为(1.269±0.167)ng/细胞、(0.923±0.119)ng/细胞、(0·766±0.085)ng/细胞及(0.682±0.063)ng/细胞,与CTGF组比较呈浓度依赖性降低(P<0.01)。④50ng/LCTGF组的心肌细胞p-ERK1/2表达强度明显高于对照组,5μmol/L、10μmol/L、25μmol/L、50μmol/L拉西地平干预组心肌细胞p-ERK1/2的表达强度与CTGF组比较呈浓度依赖性降低。心肌细胞t-ERK1/2的表达在各组之间没有明显的差异。结论拉西地平可抑制CTGF诱导的心肌细胞肥大,其作用机制可能与ERK1/2磷酸化有关。
Objective To observe the effect of lacidipine on cardiac myocyte hypertrophy induced by connective tissue growth factor (CTGF) in rats and its relationship with extracellular signal-regulated kinase 1/2 (ERK1 / 2). Methods Cardiomyocytes from neonatal SD rats were used as experimental models. The effect of lacidipine on CTGF was evaluated by image analysis, 3H-leucine incorporation, Coomassie brilliant blue staining and Western blotting. Inhibitory effect of induced cardiomyocyte hypertrophy and its relationship with ERK1 / 2. Results ① After treated with 50ng / L CTGF for 24 hours, the surface area of myocardial cells was (1812.52 ± 168.73) μm 2, which was significantly higher than that of the control group (689.31 ± 96.58) μm 2 (P <0.01) Compared with CTGF group, the myocardial cell surface area of lacidipine intervention group was (1476.52 ± 156.73) μm2, (1120.39 ± 149.68) μm2, (926.10 ± 101.44) μm2 and (739.81 ± 91.55) μm2, respectively) <0.01). ② 3H-leucine incorporation rate of cardiomyocytes was (2368.72 ± 122.45) cpm / well after treated with 50ng / L CTGF for 24 h, significantly increased compared with control group (950.26 ± 89.43) cpm / well (P <0.01) ). The 3H-leucine incorporation rates of cardiomyocytes in the 5, 10, 25 and 50 μmol / L rapamycin-treated groups were (2023.12 ± 106.15) cpm / well, (1629.15 ± 103.46) cpm / well and (1302.19 ± 98.53) cpm / well and (1055.72 ± 90.96) cpm / well in a dose-dependent manner compared with CTGF group (P <0.01). (3) Compared with control group (0.622 ± 0.068) ng / cell, the content of cardiomyocyte protein was (1.692 ± 0.203) ng / The protein levels of cardiomyocytes in the 25 and 50 μmol / L rapamycin treated groups were (1.269 ± 0.167) ng / cell, (0.923 ± 0.119) ng / cell, (0.766 ± 0.085) ng / 0.063) ng / cells in a dose-dependent manner compared with CTGF group (P <0.01). The expression of p-ERK1 / 2 in 50ng / LCTGF group was significantly higher than that in control group. The expression of p-ERK1 / 2 in 5μmol / L, 10μmol / L, 25μmol / L and 50μmol / Compared with the CTGF group in a concentration-dependent reduction. Cardiac myocyte t-ERK1 / 2 expression was not significantly different between the groups. Conclusion Lacisidipine can inhibit CTGF-induced cardiomyocyte hypertrophy and its mechanism may be related to ERK1 / 2 phosphorylation.