miR-34在小鼠心脏组织中的表达和功能研究  

Function and expression pattern of miR-34 in mice heart

作　　者：陈锡峰[1] 陈飞[1] 肖波[1,2] 黄安庆 

机构地区：[1]苏州大学基础医学与生物科学学院,江苏苏州215123 [2]盐城工学院环境科学与工程学院,江苏盐城224051

出　　处：《西部医学》2014年第10期1264-1267,共4页Medical Journal of West China

基　　金：江苏省博士后科研资助计划第一批资助项目(1301054C)

摘　　要：目的探讨miR-34家族中miR-34a、miR-34b、miR-34c在心脏组织的表达情况,确定这个家族中哪一个microRNA是主要表达形式;并比较miR-34家族在年轻小鼠心脏和老年小鼠心脏的表达差异,检测其中哪一个成员对由衰老引起的心脏功能退化具有敏感性,可作为心脏功能的检测标志。方法提取小鼠心脏总RNA,对miRNA采用加尾法,反转成cDNA。使用RT-PCR的方式检测miR-34a、miR-34b、miR-34c引物的特异性。随后使用实时荧光定量PCR的方法检测miR-34a、miR-34b、miR-34c三个基因在心脏细胞中的表达差异。同时比较年轻小鼠和老年小鼠心脏组织中的miR-34a、miR-34b、miR-34c表达量。结果提取的总RNA经检测符合要求,反转成cDNA后分别用miR-34a、miR-34b、miR-34c的荧光定量PCR引物检测,确认引物特异性极好,只有单一条带,可以进行下游荧光定量实验。在小鼠心脏组织中miR-34a的表达量较miR-34b和miR-34c高,说明miR-34a在miR-34家族中起主导作用。在老年小鼠心脏中miR-34a的表达量高于年轻小鼠(2.5倍)。miR-34b在老年小鼠心脏中也有部分上调(1.4倍),而miR-34c在年轻小鼠和老年小鼠心脏中无明显变化。结论在miR-34家族中,miR-34a的表达起主导地位,表达量高于miR-34b、miR-34c。随着年龄增加,miR-34a、miR-34b在老年小鼠心脏的表达量比年轻小鼠心脏要高,其中miR-34a2.5倍的增幅明显大于miR-34b1.4倍的增幅。故可将miR-34a作为心脏由衰老引起的功能退化的标志基因,也可作诊断心脏功能的重要指标。Objective To investigate the expression level of miR-34 family in heart to confirm which one is dominant and compare miR-34 in young and old mice heart to detect which member is sensitive to heart aging.Methods The total RNA and add the poly A tail to mature miRNA were extracted.The miRNA was reverse-transcribed into cDNA and detected the specificity of q-PCR primer.The expression levels of miR-34a,miR-34b and miR-34c were detected by realtime quantitive PCR in heart tissue.Meanwhile,we also compared the expression levels of miR-34a、miR-34b、miR-34c in the young mice with the old mice.Results After miRNA was reverse-transcribed into cDNA,we confirm the specificity of q-PCR primer.miR-34a of mice heart was dominant in miR-34 family.miR-34a was nearly 2.5 times higher in the young mice heart than it in the old mice heart and miR-34b was also up-regulated 1.4 times.However there was no significant difference of miR-34c in young and old mice.Conclusion miR-34a,the dominant in miR-34 family,is higher than miR-34b and miR-34c.miR-34a and miR-34b are both increased in old mice.But miR-34a was increased more than miR-34b in old mice heart compared with young mice.miR-34a can be regarded as a gene marker of heart aging for detecting the function of heart.

关 键 词：miR-34 心脏 衰老 

分 类 号：R393[医药卫生—基础医学]