目的研究心脏能量代谢脂肪酸β氧化过程中关键脱氢酶之一的短链脂酰辅酶A脱氢酶(short-chain acly-coen-zyme A dehydrogenase,SCAD)与心肌肥大之间的关系,探索治疗心血管疾病的新靶点。方法利用自发性高血压大鼠(spontaneous hypertens...目的研究心脏能量代谢脂肪酸β氧化过程中关键脱氢酶之一的短链脂酰辅酶A脱氢酶(short-chain acly-coen-zyme A dehydrogenase,SCAD)与心肌肥大之间的关系,探索治疗心血管疾病的新靶点。方法利用自发性高血压大鼠(spontaneous hypertension rats,SHR)和异丙肾上腺素皮下注射两种心肌肥大动物模型,以及苯肾上腺素(PE)刺激的心肌细胞肥大模型,检测SCAD在蛋白水平和mRNA水平上表达量的改变;并采用siRNA对SCAD进行干扰,观察对肥大分子指标的影响。结果以上动物模型和细胞模型中SCAD的蛋白表达水平都有明显的下降,mRNA水平表达量也有一定程度的下降。针对SCAD的siRNA干扰后可导致心肌细胞SCAD的表达明显下降,而肥大标记因子ANF、BNP的表达明显上调,进一步明确了SCAD与心肌细胞肥大的关系。结论心肌肥大时SCAD的蛋白水平和mRNA水平均明显下降,干扰SCAD后心肌细胞肥大指标上升,说明SCAD的下降可能参与心肌肥大的病理过程,上调SCAD有可能成为干预心肌肥大的重要环节之一。展开更多
NADP(H) dependent retinol dehydrogenase/reductase (NRDR) was an important retinoic acid synthase, which was first purified from rabbit liver in 1997. In order to study the function of the NRDR gene,the full length cDN...NADP(H) dependent retinol dehydrogenase/reductase (NRDR) was an important retinoic acid synthase, which was first purified from rabbit liver in 1997. In order to study the function of the NRDR gene,the full length cDNA of bovine NRDR was cloned. According to the conserved sequences of human, mouse and rabbit NRDR cDNA, a pair of primers was designed to amplify a 294 bp DNA fragment of bovine liver NRDR, and then the full length of NRDR cDNA (AF487454) was cloned by using 3′ RACE and 5′ RACE. All the cloned NRDR proteins consist of 260 amino acid residues and showed high identity among them. The tri peptide of human, mouse and rabbit NRDR C end was SRL and that of bovine NRDR C end was SHL, but both were considered to be peroxisomal target signal 1 (PTS1). RT PCR demonstrated that NRDR gene was expressed in liver, heart, lung, kidney, stomach and intestine, and was not found in pancreas, muscle, artery and skin. The full length bovine NRDR cDNA has been successfully cloned and the sequence was analyzed. It provided a reliable foundation to investigate the biological function of this protein.展开更多
家蚕质型多角体病毒(Bombyx mori cytoplasmic polyhedrosis virus,BmCPV)是家蚕的重要病毒病原之一,往往给养蚕业生产造成极大危害。我们以前的研究运用基因芯片技术在感染质型多角体病毒的家蚕中肠中鉴定出一个差异表达的3-羟酰辅酶A...家蚕质型多角体病毒(Bombyx mori cytoplasmic polyhedrosis virus,BmCPV)是家蚕的重要病毒病原之一,往往给养蚕业生产造成极大危害。我们以前的研究运用基因芯片技术在感染质型多角体病毒的家蚕中肠中鉴定出一个差异表达的3-羟酰辅酶A脱氢酶蛋白基因(Bombyx mori3-hydroxyacyl-CoA dehyrogenase protein gene-Bm3HAD)。本研究利用cDNA末端快速扩增技术(RACE)克隆了该基因,其全长cDNA序列为1168bp,包含一个83bp5'端非翻译区序列(5'-UTR)、一个930bp的开放阅读框(ORF)和一个155bp的3'端非翻译区序列(3'-UTR);基因结构分析发现该基因由5个外显子和4个内含子组成。RT-PCR结果显示该基因在家蚕中肠、脂肪体、血液、丝腺及生殖体中均有表达。荧光定量PCR结果表明该基因在BmCPV感染初期为上调表达,随着病毒感染的进展,该基因的表达水平逐渐降低,并转变为下调表达。研究结果为进一步研究BmCPV对家蚕致病的分子机制提供了有益的信息。展开更多
文摘NADP(H) dependent retinol dehydrogenase/reductase (NRDR) was an important retinoic acid synthase, which was first purified from rabbit liver in 1997. In order to study the function of the NRDR gene,the full length cDNA of bovine NRDR was cloned. According to the conserved sequences of human, mouse and rabbit NRDR cDNA, a pair of primers was designed to amplify a 294 bp DNA fragment of bovine liver NRDR, and then the full length of NRDR cDNA (AF487454) was cloned by using 3′ RACE and 5′ RACE. All the cloned NRDR proteins consist of 260 amino acid residues and showed high identity among them. The tri peptide of human, mouse and rabbit NRDR C end was SRL and that of bovine NRDR C end was SHL, but both were considered to be peroxisomal target signal 1 (PTS1). RT PCR demonstrated that NRDR gene was expressed in liver, heart, lung, kidney, stomach and intestine, and was not found in pancreas, muscle, artery and skin. The full length bovine NRDR cDNA has been successfully cloned and the sequence was analyzed. It provided a reliable foundation to investigate the biological function of this protein.
文摘家蚕质型多角体病毒(Bombyx mori cytoplasmic polyhedrosis virus,BmCPV)是家蚕的重要病毒病原之一,往往给养蚕业生产造成极大危害。我们以前的研究运用基因芯片技术在感染质型多角体病毒的家蚕中肠中鉴定出一个差异表达的3-羟酰辅酶A脱氢酶蛋白基因(Bombyx mori3-hydroxyacyl-CoA dehyrogenase protein gene-Bm3HAD)。本研究利用cDNA末端快速扩增技术(RACE)克隆了该基因,其全长cDNA序列为1168bp,包含一个83bp5'端非翻译区序列(5'-UTR)、一个930bp的开放阅读框(ORF)和一个155bp的3'端非翻译区序列(3'-UTR);基因结构分析发现该基因由5个外显子和4个内含子组成。RT-PCR结果显示该基因在家蚕中肠、脂肪体、血液、丝腺及生殖体中均有表达。荧光定量PCR结果表明该基因在BmCPV感染初期为上调表达,随着病毒感染的进展,该基因的表达水平逐渐降低,并转变为下调表达。研究结果为进一步研究BmCPV对家蚕致病的分子机制提供了有益的信息。
