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目的通过肾上腺素能受体激动剂和拮抗剂干预体外培养的人心房成纤维细胞,研究β受体在成纤维细胞增殖分泌中的作用及机制。方法:取体外循环患者右心耳组织行成纤维细胞分离培养、鉴定。实验分4组,对照组(CON组)、异丙肾上腺素组(ISO组)、ISO+阿替洛尔组(ISO+ATE组)、ISO+卡维地洛组(ISO+CAR组);干预后检测上清液中羟脯氨酸含量、细胞增殖、β受体mRNA及pPKA R2蛋白的表达。结果:1,ISO+CAR组和ISO组的细胞增殖率分别为11.3%和33.8%,P<0.01;其上清液中羟脯氨酸含量分别为6.32±1.30μg/ml和8.68±1.80μg/ml,P<0.01;2,β受体亚型mRNA的表达:β2AR/GAPDH在ISO+CAR和ISO组分别为0.52和0.66,P<0.01;而ATE+ISO组和ISO组分别为:0.63和0.66,P>0.05。3,pPKA蛋白表达:ISO+CAR组同ISO组相比,pPKA R2/β-actin分别是0.60和0.81,P<0.01。结论:ISO能刺激体外培养的人心房成纤维细胞增殖和分泌胶原;非选择性β受体拮抗剂卡维地洛可抑制成纤维细胞增殖及分泌,其机制与β2AR-pPKA R2途径有关。
OBJECTIVE: To investigate the effect and mechanism of β receptor on the proliferation and secretion of fibroblasts by adrenergic receptor agonist and antagonist intervention in human atrial fibroblasts cultured in vitro. Methods: Right atrial appendage of patients undergoing cardiopulmonary bypass was isolated and cultured in vitro. The experimental group was divided into 4 groups: control group (CON group), isoproterenol group (ISO group), ISO + atenolol group (ISO + ATE group) and ISO + carvedilol group The content of hydroxyproline in the supernatant, cell proliferation, β receptor mRNA and pPKA R2 protein expression were detected. The cell proliferation rates in ISO + CAR group and ISO group were 11.3% and 33.8%, respectively, P <0.01. The contents of hydroxyproline in the supernatant were 6.32 ± 1.30μg / ml and 8.68 ± 1.80μg / ml, P <0.01; 2, mRNA expression of beta receptor subtype: β2AR / GAPDH was 0.52 and 0.66 respectively in ISO + CAR and ISO groups, P <0.01; while ATE + ISO and ISO groups were 0.63 And 0.66, P> 0.05. 3, pPKA protein expression: Compared with ISO group, pPKA R2 / β-actin in ISO + CAR group was 0.60 and 0.81, P <0.01. CONCLUSION: ISO can stimulate the proliferation and secretion of collagen in human atrial fibroblasts cultured in vitro. Carvedilol, a non-selective β-receptor antagonist, can inhibit the proliferation and secretion of fibroblasts. The mechanism is related to β2AR-pPKA R2 pathway.