OMT抑制钙调神经磷酸酶改善醛固酮诱导的心肌成纤维细胞增殖

OMT Inhibiting Calmodulin Neurophosphatase in Aldosterone Induced Proliferation of Myocardial Fibroblasts

目的:观察氧化苦参碱(OMT)对醛固酮(ALD)诱导心肌成纤维细胞(CFs)增殖分化模型的干预作用,探讨其对钙调神经磷酸酶(Ca N)表达的影响。方法:采用胰酶消化法、差速贴壁法获得体外培养1～3 d新生SD乳鼠CFs,建立ALD诱导CFs增殖模型,采用波形蛋白(Vimentin)免疫细胞化学法鉴定CFs;将鉴定并传代的2～3代CFs分为对照组(等量蒸馏水)、ALD组(1×10^(-7)mol/L)、OMT组(1×10^(-7)mol/L ALD及7. 56×10-4mol/L OMT)及Ca N I组[1×10^(-7)mol/L ALD及Ca N抑制剂环孢素A(Cs A1)],作用24 h;采用实时荧光定量PCR(qRT-PCR)测定Ca N mRNA水平,Western blot法测定Ca N蛋白表达水平。结果:Vimentin鉴定分离培养的细胞结果阳性,提示分离细胞为CFs; ALD诱导24 h时,与对照组比较,ALD组Ca N mRNA及Ca N蛋白的表达水平升高(P <0. 01);与ALD组比较,OMT组Ca N mRNA及Ca N蛋白的表达水平降低(P <0. 01)。结论:OMT能够抑制ALD诱导的CFs增殖,其机制可能与抑制Ca N基因表达有关。

Objective： To investigate the effects of oxymatrine （OMT） on the proliferation and differentiation of myocardial fibroblasts induced by aldosterone （ALD） and explore the effect of OMT on the expression of calmodulin （CAN）. Methods： CFs of newborn SD mice cultured for 1 -3 days in vitro was obtained and CFs Proliferation Model induced by ALD was established by the method of trypsin digestion and differential adhension. CFs was authenticated by vimentin inmmnocytochemistry. Generation 2 and 3 CFs were divided into four groups： control group, ALD group （1 × 10^-7 mol/L） , OMT group（ 1× 10^-7 mol/L ALD and 7.56 × 10^ -4 mol/L OMT） and CaN I group [1 × 10^-7 mol/L ALD and CaN inhibitor cyclosporine A（CsA1 ）]. CaN mRNA level was deteermined by Real-time fluorescence quantitative PCR （qRT-PCR） , and CaN protein expression was detennined by Western blot. Results： The result of cells isolated and cultured by identification of vimentin was positive and indicated that the isolated cells were CFs. The expression of CaN mRNA and CaN protein in ALD group was higher than that in control group at 24 h after induction（P 〈0.01）. The expression of CaN mRNA and CaN protein in OMT group was lower than that in ALD group（P 〈0.01 ）. Conclusion： OMT can inhibit the proliferation of CFs induced by ALD, and its mechanism may be related to the inhibition of CaN gene expression.