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  

Atorvastatin can prevent transformation of aortic valvular myofibroblast to an osteoblast-like phenotype

作　　者：沈迎念[1] 张莉伟[1] 辜刚健 李永胜[1] 

机构地区：[1]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,湖北武汉430030

出　　处：《中国医院药学杂志》2015年第12期1057-1061,共5页Chinese Journal of Hospital Pharmacy

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)

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(P<0.05),加入Ator后,细胞的凋亡速度明显减低(P<0.05)。ox-LDL组α-SMA的表达明显增加(P<0.05),成为活性细胞,加用Ator并未对α-SMA蛋白产生影响(P>0.05)。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(P<0.01),加入Ator后,BMP2、ALP的表达明显下降(P<0.05)。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。OBJECTIVE To study the effectsrole of atorvastatin on apoptosis and bone related proteins expression of oxidized lowdensity lipoprotein（ ox-LDL） treated aortic valvular myofibroblast,and explore its potential therapeutic mechanisms on preventing the progression of aortic valve calcification. METHODS Valvular myofibroblastes isolated from porcine aortic valvulare leaflets waere treated with ox-LDL（25 mg·L^-1） in the presence or absence of atorvastatin（50 mg·L^-1） and PBS or Atorvastatin（50 mg·L^-1 alone as controls respectively. Cells were then harvested at different time points（24,48,72 h）. Cell apoptosis was checked by flow cytometry.Protein and gene expression levels of α-smooth muscle actin（ α-SMA） and bone related proteins,bone morphogenetic protein 2（ BMP2）and alkaline phosphatase（ ALP） were analyzed by Western-blot and real-time PCR. RESULTS In comparisoned with the control group,ox-LDL treatment could time-dependently activate aortic valvular myofibroblasts by promoting its apoptosis and in the meanwhile inducing its expression of bone related proteins,BMP2 and ALP rather than,but not α-SMA,while Atorvastatin treatment couldan suppress cell apoptosis and its expression of BMP2 and ALP. CONCLUSION ox-LDL can cause aortic valve calcification by promoting the differentiation,proliferation,apoptosis and osteoblast-like phenotype transformation of aortic valvular myofibroblasts,whereas Atorvastatin can into a certain extent reverse this process,suggesting its potential therapeutic effects against for aortic valve calcification.

关 键 词：阿托伐他汀 OX-LDL 成肌纤维细胞 凋亡 瓣膜钙化 

分 类 号：R972.6[医药卫生—药品]