钙调神经磷酸酶在去甲肾上腺素诱导的心肌成纤维细胞活化中的作用

Calcineurin participates in regulating norepinephrine-induced activation of cardiac fibroblasts

目的:研究钙调神经磷酸酶对去甲肾上腺素诱导的心肌成纤维细胞增殖及表型转化的调节作用。方法:酶消化法原代培养SD大鼠乳鼠的心肌成纤维细胞(CF),运用细胞计数法和MTT法测定CFs的增殖反应,Western blot测定钙调神经磷酸酶(CaN)、α-平滑肌肌动蛋白(α-SMA)的蛋白表达,定磷法检测CaN活性。结果:去甲肾上腺素可以明显促进CFs增殖及表型转化,促进CaN蛋白的表达及活性。酚妥拉明和环孢菌素A均可抑制去甲肾上腺素诱发的CFs增殖,环孢菌素A抑制去甲肾上腺素诱发的α-SMA及CaN的表达和活性。结论:钙调神经磷酸酶参与去甲肾上腺素诱导的心肌成纤维细胞增殖及表型转化。

Objective：To investigate calcineurin in the regulation of cardiac fibroblasts proliferation and differentiation induced by norepinephrine.Method：Primary cultures of neonatal rat cardiac fibroblasts（CFs）were enzymatically isolated from the hearts of Sprague Dawley.Proliferation of CFs was determined using cell counts and MTT assay.The protein of calcineurin（CaN）and alpha-smooth muscle actin（α-SMA）were assayed with immunoblotting.CaN activity was assessed with Free inorganic phosphate content analysis.Result：Norepinephrine（NE）increased CFs proliferation,cell number,differentiation,promoted CaN protein expression and activated CaN.Phentolamine（Phen）and cyclosporin A（CsA）decreased NE-induced proliferation,cell number,differentiation.CsA also decreased NE-induced CaN levels and activity.Conclusion：CaN is involved in regulating NE-induced proliferation and differentiation of cardiac fibroblasts.