坎地沙坦抑制血管紧张素Ⅱ诱导的大鼠胸主动脉平滑肌A7r5细胞增殖和迁移及机制

Candesartan inhibits the angiotensin Ⅱ-induced proliferation and migration of rat thoracic aortic smooth muscle A7r5 cells and its mechanism

目的探讨坎地沙坦对血管紧张素Ⅱ(AngⅡ)诱导的血管平滑肌细胞增殖和迁移及对连接子蛋白43(Cx43)的影响和机制。方法体外培养A7r5大鼠胸主动脉平滑肌细胞,随机分为对照组、 AngⅡ组及AngⅡ联合坎地沙坦组。CCK-8法检测A7r5细胞的增殖, Transwell^(TM)小室实验和划痕实验检测A7r5细胞的迁移能力;免疫荧光细胞化学技术检测A7r5细胞Cx43的表达和分布, Western blot法检测A7r5细胞Cx43、骨桥蛋白(OPN)、增殖细胞核抗原(PCNA)、磷酸化的丝裂原激活胞外调节激酶1/2(p-MEK1/2)及磷酸化的胞外信号调节激酶1/2(p-ERK1/2)的蛋白水平。结果与对照组相比, AngⅡ组细胞增殖和迁移能力增强;与单用AngⅡ组相比, AngⅡ组联合坎地沙坦组的细胞增殖和迁移能力随坎地沙坦浓度增加而降低; Cx43表达在A7r5细胞膜和核膜,与对照组相比, AngⅡ组Cx43表达增强, Cx43、 OPN、 PCNA、 p-MEK1/2及p-ERK1/2蛋白水平均较AngⅡ组明显增加, AngⅡ联合坎地沙坦组Cx43表达及各蛋白水平均明显降低。结论坎地沙坦可降低AngⅡ诱导平滑肌细胞的增殖和迁移,可能与抑制Cx43及MEK/ERK表达有关。

Objective To investigate the effect of candesartan on angiotensin Ⅱ(Ang Ⅱ)-induced proliferation and migration of vascular smooth muscle cells and its effect on connexin 43(Cx43). Methods A7r5 cells were cultured in vitro and randomly divided into control group, AngⅡ group and AngⅡ combined with candesartan group. Cell viability was detected by CCK-8 assay;the migration and invasion ability of A7r5 cells were measured by wound-healing and TranswellTM assay;the expression and distribution of Cx43 on A7r5 cells were detected by immunofluorescence assay. Cx43, osteopontin(OPN), proliferating cell nuclear antigen(PCNA), p-MEK1/2 and p-ERK1/2 protein levels of A7r5 cells were detected by Western blot analysis. Results Compared with the control group, the AngⅡ group had higher cell proliferation and migration ability, and the AngⅡ combined with candesartan group had a lower concentration than the AngⅡ group. Cx43 was expressed in the A7r5 cell membrane and nuclear membrane. The expression of Cx43 were enhanced in the AngⅡ group than in the control group, and the expressions of Cx43, OPN, PCNA, p-MEK1/2 and p-ERK1/2 significantly increased compared with AngⅡ. The expression of protein significantly decreased in AngⅡ combined with candesartan group. Conclusion Candesartan can reduce the proliferation and migration of smooth muscle cells induced by AngⅡ, and its mechanism may be related to Cx43 and MEK/ERK signaling pathways.