CAJ | 학술논문

目的：构建并鉴定心肌组织特异性高表达核仁素（ Ncl ）的转基因小鼠，为从整体水平上研究核仁素对心肌功能的影响及其发挥心肌保护作用的机制提供动物模型。方法：采用基因重组法将小鼠核仁素基因的全长cDNA插入含心肌细胞特异性启动子的Alpha-MyHC clone 26载体，构建Alpha-MyHC clone 26-Ncl重组载体，通过双酶切及基因测序鉴定其正确性；采用PCR鉴定阳性小鼠并用Western blotting法观察转基因小鼠中核仁素的表达情况，采用组织切片及HE染色法观察转基因小鼠心肌组织形态；测量心脏重量指数和左室压最大上升速率，观察核仁素在心肌组织高表达后对小鼠心脏形态功能是否有影响。结果： PCR法观察显示获得4只阳性小鼠（51、52、56和86，其中52和86系繁殖良好），Western blotting结果表明转基因小鼠的核仁素表达量明显高于野生鼠，HE染色等结果表明核仁素对心肌形态学、心脏/体重比值及心功能无明显影响。结论：成功制备了心肌组织特异性高表达核仁素的转基因小鼠，与野生型小鼠相比，转基因小鼠心肌形态及心功能无明显差异。
목적：구건병감정심기조직특이성고표체핵인소（ Ncl ）적전기인소서，위종정체수평상연구핵인소대심기공능적영향급기발휘심기보호작용적궤제제공동물모형。방법：채용기인중조법장소서핵인소기인적전장cDNA삽입함심기세포특이성계동자적Alpha-MyHC clone 26재체，구건Alpha-MyHC clone 26-Ncl중조재체，통과쌍매절급기인측서감정기정학성；채용PCR감정양성소서병용Western blotting법관찰전기인소서중핵인소적표체정황，채용조직절편급HE염색법관찰전기인소서심기조직형태；측량심장중량지수화좌실압최대상승속솔，관찰핵인소재심기조직고표체후대소서심장형태공능시부유영향。결과： PCR법관찰현시획득4지양성소서（51、52、56화86，기중52화86계번식량호），Western blotting결과표명전기인소서적핵인소표체량명현고우야생서，HE염색등결과표명핵인소대심기형태학、심장/체중비치급심공능무명현영향。결론：성공제비료심기조직특이성고표체핵인소적전기인소서，여야생형소서상비，전기인소서심기형태급심공능무명현차이。
AIM:To construct myocardium-specific nucleolin ( Ncl) transgenic mice and to provide an animal model for the studies of the myocardial protection of nucleolin .METHODS:To create nucleolin transgenic mice , a myo-cardium-specific expression plasmid of nucleolin ( Alpha-MyHC clone 26-Ncl) was constructed .The gene type of transgenic mice was identified by PCR and the nucleolin protein level was tested by Western blotting .The myocardium morphology , heart weight index (HWI) and left ventricular pressure maximum rise rate were observed in nucleolin transgenic (TG) mice and wild-type ( WT) mice.RESULTS:We gained 4 transgenic mice (51, 52, 56 and 86 lines, only 52 line and 86 line were eugonic) by PCR.Western blotting analysis showed the expression of nucleolin up-regulated specifically in the myocardium .However , the myocardium morphology , HWI and left ventricular pressure maximum rise rate in the nucleolin transgenic mice were similar to those in the wild-type mice.CONCLUSION:We constructed myocardium-specific nucleo-lin transgenic mice successfully .