抑制LncRNA PANDA表达对人心脏瓣膜间质细胞钙化的改善作用及其机制  

Effect of inhibition of LncRNA PANDA expression on calcification of human heart valve stromal cells and its mechanism

作　　者：凌秋洋 赵苗茁 王广涛 杨志宏 刘叶红 叶挺 宗刚军 LING Qiuyang;ZHAO Miaozhuo;WANG Guangtao;YANG Zhihong;LIU Yehong;YE Ting;ZONG Gangjun(Department of Geriatric Rehabilitation,Qingdao Special Servicemen Recuperation Center of PLA Navy,Qingdao 266000,China;不详)

机构地区：[1]中国人民解放军海军青岛特勤疗养中心老年病康复科,山东青岛266000 [2]中国人民解放军海军第九七一医院心内科 [3]中国人民解放军联勤保障部队第九〇四医院心内科

出　　处：《山东医药》2023年第23期43-46,共4页Shandong Medical Journal

基　　金：江苏省自然科学基金资助项目(BK20201139)。

摘　　要：目的观察抑制长链非编码RNA PANDA(LncRNA PANDA)表达对人心脏瓣膜间质细胞(hVICs)钙化的影响,并探讨其机制是否与核酸转录因子YA(NF-YA)、凋亡相关蛋白Bax、锚定蛋白2(Notch2)、骨形态发生蛋白2(BMP-2)有关。方法将体外培养的hVICs分为三组,正常对照组加入间质细胞培养基+10%胎牛血清,培养7 d;钙化组加入钙化诱导液,培养7 d。PANDA-siRNA组加入钙化诱导液培养3 d后,采用Lipofectamine^(TM)3000转染试剂盒转染PANDA-siRNA,再培养4 d。取各组分组培养7 d的细胞,采用实时荧光定量PCR法检测LncRNA PANDA相对表达量,光学显微镜观察细胞形态,Western blotting法检测NF-YA、Bax、Notch2、BMP-2蛋白相对表达量,ELISA法检测细胞上清液BMP-2水平。结果PANDA-siRNA组、正常对照组、钙化组细胞LncRNA PANDA相对表达量及上清液BMP-2蛋白水平均依次升高,组间两两比较P均<0.05。PANDA-siRNA组细胞结构清晰,呈条索状生长,生长稍缓慢;钙化组细胞成骨样改变,细胞聚集,呈网状结构;正常对照组细胞贴壁生长,生长迅速。与正常对照组比较,钙化组细胞NF-YA、Bax、Notch2、BMP-2蛋白相对表达量均升高,PANDA-siRNA组均降低(P均<0.05)。结论诱导钙化后的hVICs LncRNA PANDA表达升高,抑制LncRNA PANDA表达可减轻其成骨样改变,其机制可能与降低NF-YA、Bax、Notch2、BMP-2表达有关。Objective To observe the effect of inhibition of long non-coding RNA PANDA(LncRNA PANDA)ex⁃pression on the calcification of human heart valve stromal cells(hVICs)and to investigate whether its mechanism is related to the the expression of nucleic acid transcription factor YA(NF-YA),apoptosis-related protein(Bax),anchor protein 2(Notch2),and bone morphogenic protein-2(BMP-2).The hVICs cultured in vitro were divided into three groupsin Methods The hVICs cultured in vitro were divided into three groups:the normal control group was cultured with intersti⁃tial cell medium+10%FBS for 7 d;the calcified group was supplemented with the calcification induction solution and was cultured for 7 d;the PANDA-siRNA group was incubated for 3 d with the calcification induction solution,and then was transfected with PANDA-siRNA by Lipofectamine 3000 transfection kit and was cultured for another 4 d.Cells cultured in groups for 7 d were selected.The relative expression of LncRNA PANDA was measured by real-time fluorescent quantita⁃tive PCR,cell morphology was observed by light microscope,the relative expression levels of NF-YA,Bax,Notch2,and BMP-2 proteins were detected by Western blotting,and BMP-2 level in cell supernatant was detected by ELISA.Results The relative expression levels of LncRNA PANDA and BMP-2 protein in the PANDA-siRNA group,normal control group,and calcified group increased successively,with statistically significant difference between groups(all P<0.05).The cells in the PANDA-siRNA group had clear cell structure with cord-like growth and slightly slow growth,the cells in the calci⁃fied group had osteogenic changes with cell aggregation and network structure,and the cells in the normal control group grew adhering to the wall and grew rapidly.Compared with the normal control group,the relative protein expression levels of NF-YA,Bax,Notch2 and BMP-2 all increased in the calcified group,and decreased in the PANDA-siRNA group(all P<0.05).Conclusion LncRNA PANDA expression increases in hVICs after induction of

关 键 词：长链非编码RNA PANDA 核酸转录因子YA 凋亡相关蛋白 骨形态发生蛋白2 人心脏瓣膜间质细胞 瓣膜钙化 心脏瓣膜病 

分 类 号：R542.5[医药卫生—心血管疾病]