微小RNA-22通过抑制去乙酰化转移酶6的表达在主动脉瓣钙化的发生发展中的作用  

The role of microRNA-22 in the occurrence and development of aortic valve calcification by inhibiting the expression of sirtuin 6

作　　者：谢晓勇[1] 冯旭[1] 贺洲 朱晓莉 温大帅 Xie Xiaoyong;Feng Xu;He Zhou;Zhu Xiaoli;Weng Dashuai(Department of Cardiac Surgery,the First Affiliated Hospital of Guangxi Medical University,Nanning 530000,China)

机构地区：[1]广西医科大学第一附属医院心脏外科,南宁530000

出　　处：《中华实验外科杂志》2022年第8期1476-1479,共4页Chinese Journal of Experimental Surgery

基　　金：广西自然科学基金项目(桂科记字[2019]185号)。

摘　　要：目的探讨微小RNA-22(miR-22)对瓣膜间质细胞成骨分化的作用及其在主动脉瓣钙化发生发展中的作用。方法以主动脉瓣置换术中切除的钙化性主动脉瓣疾病15例, 其中钙化主动脉瓣组织作为实验组, 正常主动脉瓣组织为对照组;苏木精-伊红(HE)染色、VonKossa法观察瓣膜钙化程度;实时定量反转录聚合酶链反应(RT-qPCR)、免疫荧光试验检测瓣膜组织中miR-22的表达变化和蛋白表达水平。组间比较采用方差分析。结果 HE染色以及Von Kossa钙沉积染色提示实验组中有大量羟基磷灰石沉积;免疫组织化学染色结果显示实验组Runx相关转录因子2(Runx2)、骨钙素(OCN)的阳性表达。相较对照组, 实验组miR-22基因表达明显升高(P<0.01), 实验组成骨相关基因Runx2高于对照组(1.39±0.02比1.09±0.01, F=4.00, P<0.01);实验组成骨相关基因OCN高于对照组(0.92±0.03比0.58±0.02, F=2.250, P<0.01), 实验组去乙酰化转移酶6(HDAC6)低于对照组(1.09±0.05比1.52±0.04, F=1.563, P<0.01)。双荧光素报告基因检测和Western blot检测结果显示miR-22过表达可以显著增加成骨相关基因的表达;过表达miR-22显著减低HDAC6的表达。结论 miR-22通过抑制HDAC6的表达促进Runx2、OCN的表达和活性, 瓣膜间质细胞成骨分化, 参与主动脉瓣膜钙化进程。Objective To investigate the effect of microRNA-22(miR-22)on the osteogenic differentiation of valve interstitial cells and its role in the occurrence and development of aortic valve calcification.Methods Totally,15 patients with calcified aortic valve disease resected during aortic valve replacement were selected,of which calcified aortic valve tissue was used as the experimental group and normal aortic valve tissue as the control group.Hematoxylin and eosin(HE)staining and VonKossa method were used to observe the degree of valve calcification.Real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)and immunofluorescence assays were used to detect the expression changes and protein expression levels of miR-22 in valve tissue.Results HE staining and Von Kossa calcium deposition staining suggested that there was a large amount of hydroxyapatite deposition in the experimental group.The immunohistochemical staining results showed positive expression of Runx2 and OCN in the experimental group.The miR-22 gene expression in the experimental group was significantly higher than in the control group(P<0.01).Runx2,a bone related gene in the experimental group,was higher than that in the control group(1.39±0.02 vs.1.09±0.01,F=4.00,P<0.01).The expression of the bone related gene OCN was higher in the experimental group than in the control group(0.92±0.03 vs.0.58±0.02,F=2.250,P<0.01),and histone deacetylase 6(HDAC6)was lower in the experimental group than in the control group(1.09±0.05 vs.1.52±0.04,F=1.563,P<0.01).The results of dual luciferin reporter assay and Western blotting assay showed that miR-22 overexpression could significantly increase the expression of osteogenesis related genes.Overexpression of miR-22 significantly attenuated HDAC6 expression.Conclusion MiR-22 promotes the expression and activity of Runx2 and OCN by inhibiting the expression of HDAC6,and the osteogenic differentiation of valve interstitial cells is involved in the process of aortic valve calcification.

关 键 词：微小RNA-22 主动脉瓣钙化 去乙酰化转移酶6 基因表达 

分 类 号：R542.52[医药卫生—心血管疾病]