α7烟碱型乙酰胆碱受体激动剂对TGF-β1介导的血管外膜成纤维细胞表型转化影响的体外研究
Effect of α7 nicotinic acetylcholine receptor activation on transforming growth factor β1-induced phenotypic transformation of adventitia fibroblasts studied in vitro
Received date: 2018-09-20
Online published: 2019-02-25
关键词: α7烟碱型乙酰胆碱受体; 外膜成纤维细胞; 细胞表型转化; 机制
魏坚, 高平进, 韩卫青 . α7烟碱型乙酰胆碱受体激动剂对TGF-β1介导的血管外膜成纤维细胞表型转化影响的体外研究[J]. 诊断学理论与实践, 2019 , 18(1) : 56 -60 . DOI: 10.16150/j.1671-2870.2019.01.011
Objective: To investigate the correlation between α7 nicotinic acetylcholine receptor (α7 nAchR) and phenotypic transformation of adventitial fibroblasts (AF). Methods: AF were isolated and cultured by using thoracic aorta of male Sprague-Dawley (SD) rats. For evaluating the effect of transforming growth factor-β1 (TGF-β1) on phenotypic transformation of AF, cultured AF were divided into three groups: blank control group A, group with 24 h TGF-β1 treatment, and group with 48 h TGF-β1 treatment. After treatment, real-time quantitative PCR and western blotting were used to detect the mRNA level and protein level of α7 nAchR, and the expression of a smooth muscle actin (α-SMA) and collagen Ⅰ. To further explore whether the activation of α7 nAchR can inhibit TGF-β1-induced phenotypic transformation and the underlying molecular mechanism, AF cultured in vitro were divided into three groups: blank control group B, group with 48 h TGF-β1 treatment, and group with 48 h TGF-β1 +PNU-282987 (an α7 nAchR activator) treatment. After 48 h treatment, the expression of α-SMA and collagen Ⅰ and the phosphorylation of extracellular regulated protein kinase 1/2 (Erk1/2) were detected by immunoblotting. Results: Compared with the blank control group A, TGF-β1 significantly decreased the mRNA level and protein level of α7 nAchR (P<0.05), whereas TGF-β1 induced significant increase of a-SMA and collagen Ⅰ (P<0.05). PNU-282987, an α7 nAchR agonist, inhibited TGF-β1-induced expression of α-SMA and collagen typeⅠ. Meanwhile, PNU-282987 also significantly inhibited TGF-β1-induced Erk1/2 phosphorylation. Conclusions: The inhibition of α7 nAchR is involved in TGF-β1-induced phenotypic transformation in AF, and the mechanism may be related to Erk1/2 phosphorylation.
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