阿托伐他汀抑制心脏瓣膜成肌纤维细胞向成骨细胞样表型转化

发布时间：2018-12-28 16:20

【摘要】：目的:通过观察猪主动脉瓣成肌纤维细胞在受氧化型低密度脂蛋白(ox-LDL)作用时,阿托伐他汀(Ator)对细胞凋亡的影响以及是否表达骨相关蛋白,来探讨阿托伐他汀对心脏瓣膜钙化的防治机制。方法:以原代培养的猪主动脉瓣成肌纤维细胞作为研究对象,用ox-LDL处理细胞。将瓣膜间质细胞随机分为4组:对照组(Ctrl),ox-LDL组(25 mg·L-1ox-LDL),oxLDL+Ator组(25 mg·L-1ox-LDL+50 mg·L-1Ator),Ator组(50 mg·L-1Ator)。采用流式细胞学技术测定各组细胞在24,48,72 h的凋亡变化,利用Western印迹法和Real-time PCR检测72 h各组细胞的α-平滑肌蛋白(α-smooth muscle actin,α-SMA)活性,以及骨形成蛋白2(bone morphogenetic protein 2,BMP2),碱性磷酸酶(alkaline phosphatase,ALP)的基因表达。结果:ox-LDL分别处理成肌纤维细胞24,48,72 h后,细胞显示出时间依赖性的凋亡加速(P0.05),加入Ator后,细胞的凋亡速度明显减低(P0.05)。ox-LDL组α-SMA的表达明显增加(P0.05),成为活性细胞,加用Ator并未对α-SMA蛋白产生影响(P0.05)。骨相关蛋白BMP2、ALP和mRNA在对照组和Ator组仅有低水平表达;而在ox-LDL组表达显著增加(P0.01),加入Ator后,BMP2、ALP的表达明显下降(P0.05)。结论:ox-LDL促使心脏瓣膜成肌纤维细胞分化、凋亡加速以及向成骨细胞样表型转化;Ator可以抑制ox-LDL所诱导的细胞凋亡,并对细胞的激活,以及向成骨样细胞表型分化可以起到一定的抑制作用,对心脏瓣膜钙化有防治作用。
[Abstract]:Aim: to investigate the effect of Atto vastatin (Ator) on apoptosis and the expression of bone related protein in porcine aortic valve myofibroblasts treated with oxidized low density lipoprotein (ox-LDL). To explore the prevention and treatment mechanism of Atto vastatin on cardiac valve calcification. Methods: primary cultured porcine aortic valve myofibroblasts were treated with ox-LDL. The interstitial cells were randomly divided into 4 groups: control group (Ctrl), ox-LDL group) (25 mg L-1ox-LDL), oxLDL Ator group) (25 mg L-1ox-LDL 50 mg L-1Ator), Ator group (50 mg L-1Ator). Flow cytometry was used to detect the apoptosis of the cells in each group at 24: 48 and 72 h. The activity of 伪 -smooth muscle protein (伪 -smooth muscle protein) (伪 -smooth muscle protein) was detected by Western blotting and Real-time PCR. And the gene expression of bone morphogenetic protein 2 (bone morphogenetic protein 2 and alkaline phosphatase (alkaline phosphatase,ALP). Results: after treatment of myofibroblasts with ox-LDL for 24 ~ 48 h for 72 h, the apoptosis of myofibroblasts was accelerated in a time-dependent manner (P0.05), and Ator was added to the cells. The rate of apoptosis was significantly decreased (P0.05). The expression of 伪-SMA in ox-LDL group was significantly increased (P0.05), and the expression of 伪-SMA protein was not affected by the addition of Ator (P0.05). The expression of bone related protein BMP2,ALP and mRNA was only low in the control group and Ator group, but increased significantly in the ox-LDL group (P0.01). The expression of BMP2,ALP decreased significantly after adding Ator (P0.05). Conclusion: ox-LDL can induce cardiac valve myofibroblasts to differentiate, accelerate apoptosis and transform to osteoblast like phenotype. Ator can inhibit apoptosis induced by ox-LDL, activate cells and differentiate into osteoblast-like cells, and can prevent and cure cardiac valve calcification.
【作者单位】：华中科技大学同济医学院附属同济医院急诊科/综合医疗科;
【基金】：国家自然科学基金项目(编号:81070190) 湖北省自然科学基金项目(编号:2014CFB962)
【分类号】：R965

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