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目的:研究氧化苦参碱(OMT)对醛固酮(ALD)诱导心肌成纤维细胞(CFs)增殖的作用,并分析其对p38丝裂素活化蛋白激酶(p38MAPK)表达的影响。方法:采用胰酶消化差速贴壁法分离纯化SD新生大鼠CFs做原代培养,建立ALD诱导CFs增殖模型。实验分为空白组(无血清DMEM),ALD组(1×10-7mol·L-1),OMT高、低剂量组(3.78×10-4,7.57×10-4mol·L-1)。波形蛋白免疫细胞化学法鉴定CFs。采用噻唑蓝(MTT)法分析ALD对CFs增殖的量效与时效关系和OMT的抑制作用。实时荧光定量PCR分析p38MAPK mRNA的表达。Western blot分析p-p38MAPK,p38MAPK的蛋白表达。结果:MTT结果提示1×10-7mol·L-1ALD能够诱导CFs的增殖,且在24 h时增殖作用显著,7.57×10-4mol·L-1OMT可显著抑制CFs增殖。OMT对p38MAPK mRNA的水平无影响。OMT能够抑制p-p38MAPK蛋白的表达。结论:OMT可抑制ALD诱导CFs增殖的作用,其机制可能与抑制p38MAPK磷酸化有关。
OBJECTIVE: To investigate the effect of oxymatrine (OMT) on aldosterone-induced proliferation of cardiac fibroblasts (CFs) and its effect on the expression of p38 mitogen-activated protein kinase (p38MAPK). Methods: CFs isolated and purified from SD neonatal rats by trypsin digestion and differential adherent method were used for primary culture, and ALD-induced CFs proliferation model was established. The experiment was divided into blank group (serum-free DMEM), ALD group (1 × 10-7mol·L-1), OMT high and low dose group (3.78 × 10-4,7.57 × 10-4mol·L-1). Identification of CFs by vimentin immunocytochemistry. The effect of ALD on the proliferation of CFs was analyzed by MTT method. Real-time fluorescence quantitative PCR analysis p38MAPK mRNA expression. Western blot analysis p-p38MAPK, p38MAPK protein expression. Results: The results of MTT indicated that the proliferation of CFs was induced by 1 × 10-7mol·L-1ALD, and the proliferation of CFs was significant at 24 h. 7.57 × 10-4 mol·L-1 OMT could significantly inhibit the proliferation of CFs. OMT had no effect on the level of p38MAPK mRNA. OMT can inhibit the expression of p-p38MAPK protein. Conclusion: OMT can inhibit the proliferation of CFs induced by ALD, which may be related to the inhibition of p38MAPK phosphorylation.