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目的:探讨益气温阳活血方含药血清对鼠肥大心肌细胞儿茶酚胺、钙调神经磷酸酶的影响。方法:30只Wistar大鼠分为中药组(益气温阳活血方灌胃)、卡维地洛组(卡维地洛灌胃)、空白血清组(蒸馏水灌胃)各10只,每日给药2次,共8周。末次灌胃后处死动物,分离血清,将血清经微孔滤膜灭活、滤菌后与DMEM培养基配成10%浓度。取新生乳鼠心室肌,用0.25%胰蛋白酶消化分离,经差速纯化2h,接种于10%血清DMEM培养液中,取上述细胞加入AngⅡ作用48h诱导心肌肥大后,分为正常组(正常心肌细胞加空白血清培养)、模型组(肥大心肌细胞加空白血清培养)、中药组(肥大心肌细胞加入益气温阳活血方含药血清培养)、西药组(肥大心肌细胞加卡维地洛含药血清培养)。检测各组心肌细胞内游离钙离子浓度([Ca2+]i)、儿茶酚胺(CA)、内皮素-1(ET-1)一氧化氮(No)含量、活化的T细胞核因子3(NFAT3)核蛋白活性、CaN mRNA及蛋白质表达水平。结果:与正常组比较,模型组心肌细胞[Ca2+]i、CA、ET-1含量、CaN mRNA的表达水平以及NFAT3核蛋白活性明显升高,NO含量明显降低。结论:益气温阳活血方能够明显减少心肌细胞[Ca2+]i、CA、ET-1含量,升高NO含量,抑制CaN mRNA和蛋白质的表达水平及NFAT3核蛋白活性,其机制可能与经由CaN信号通路活化抑制交感神经兴奋有关。
Objective: To investigate the effect of serum containing Yi-Yang-Yang-Huo-Xue Decoction on the catecholamine and calcineurin of rat hypertrophic cardiomyocytes. METHODS: Thirty Wistar rats were divided into Chinese medicine group (Yiqi Yangyang Huoxue Recipe), Carvedilol group (carvedilol gavage) and blank serum group (distilled water). Drug 2 times for 8 weeks. After the last gavage, the animals were sacrificed, and the serum was separated. The serum was inactivated by a microporous membrane, and the serum was filtered and mixed with DMEM medium to form a 10% concentration. The neonatal rat ventricular myocardium was isolated and digested with 0.25% trypsin, purified by differential speed for 2 h, and inoculated in 10% serum DMEM. The above cells were added to Ang II for 48 h to induce hypertrophy and divided into normal groups (normal myocardium). Cells plus blank serum culture), model group (hypertrophic cardiomyocytes plus blank serum culture), Chinese medicine group (hypertrophic cardiomyocytes were added with Yi-Yang Yang Huoxue Recipe containing serum culture), western medicine group (hypertrophic cardiomyocytes plus carvedilol containing medicine Serum culture). The intracellular free calcium concentration ([Ca2+]i), catecholamine (CA), endothelin-1 (ET-1) nitric oxide (No) content, and activated T cell nuclear factor 3 (NFAT3) nuclear protein were detected in each group. Activity, CaN mRNA, and protein expression levels. RESULTS: Compared with the normal group, the levels of [Ca2+]i, CA, ET-1, CaN mRNA, and NFAT3 nuclear protein activity in the model group were significantly increased, and the NO content was significantly decreased. Conclusion: Yi-Yun-Yang-Yang-Huo-Xue Decoction can significantly reduce the content of [Ca2+]i, CA, ET-1 in cardiomyocytes, increase the content of NO, and inhibit the expression of CaN mRNA and protein and the nuclear protein activity of NFAT3. The mechanism may be related to the CaN signal. Activation of the pathway inhibits sympathetic nerve activity.