,Similar effects on rat renal mesangial cells by expressing different fragments of adrenomedullin ge

来源 :中国药理学报(英文版) | 被引量 : 0次 | 上传用户:ylg2008asp
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
Aim: To construct pEGFP-N3 recombinant vectors carrying adrenomedullin (AM)or fragments of the AM gene, and to express AM or fragments of AM from the pEGFP-N3 recombinant vectors (pEGFP-N3-AM 1-2 and pEGFP-N3-AM 1-3) and study their biological properties on cultured rat renal mesangial cells (RMC).Methods: Total RNA of rat kidney was obtained using TriZol reagent.The cDNA was synthesized by reverse transcriptase using oligo-deoxythymidine as primer.The fragments of AM gene were then amplified by polymerase chain reaction (PCR) with specific upstream and downstream oligonucleotides.The PCR products were digested with EcoRI and BamHI and subcloned into the plasmid pEGFPN3.Facilitated by cationic liposomes, RMC were transfected with pEGFP-N3-AM1-2 or pEGFP-N3-AM1-3.After 24 h, green fluorescent protein (GFP) fluorescent images were examined with a fluorescence microscope.After 48 h, the proliferation of RMC was detected using the MTT assay, and the mRNA expression of transforming growthfactor-β1 (TGF-β1) was measured by semiquantitative PCR.Results: DNA sequence reports verified that pEGFP-N3-AM1-2, which carried the full length AM gene translation fragment (preproadrenomedullin preproAM1-185),and pEGFP-N3-AM1-3, which carried the translation fragment of preproAM [without adrenotensin (ADT, preproAM150-185)], were constructed successfully.After 24 h, green fluorescence was observed in RMC into which either pEGFP-N3-AM1-2 or pEGFP-N3-AM 1-3 was transfected, while in the control cells no fluorescence was observed.Either pEGFP-N3-AM1-2 or pEGFP-N3-AM1-3 delivery inhibited the proliferation of RMC (P<0.01) and decreased the mRNA transcription level of TGF-β1 in RMC (P<0.05).However, no significant difference was observed between the effects of pEGFP-N3-AM1-2 and pEGFP-N3-AM1-3.Conclusion: pEGFP-N3-AM1-2 and pEGFP-N3-AM1-3 were constructed successfully and were functionally expressed in RMC.Expressing the fragment of AM without ADT has similar inhibitory biological effects on RMS proliferation and TGF-β1 transcription with full length preproAM.
其他文献
在新闻标题中,有一种数学式的类型。它力避拖沓枯燥的概念语言,而代之以新颖有特点的数学式,给读者以新鲜之感,能诱发人们的兴味。这种标题有以下几种形式: 一是数字的数学
AIM: To investigate the effect of tanshinone (Tan) on the neuropathological changes induced by amyloid β-peptide1-40 (Aβ-40) injection in hippocampus in rats.
关于新闻的思维,近年来新闻界有这样一个观点,认为新闻的思维是直观思维,这种思维是区别于逻辑思维和形象思维的又一种反映客观世界的思维方式。这种大胆探索新闻规律的精神
安徽省是农业大省,又是全国农村改革的先行者,没有广大农民的小康和富裕,就没有全省的崛起和繁荣.天长市是农业大市,也是全国优质商品粮生产基地,2004年全市粮食总产已达6.3
《少年先锋》报,是少共川陕省委机关报。它是一九三三年七月六日在今达县地区通江新场坝召开了川陕省第一次团员代表大会,正式成立了少共川陕省委后,决定创办的。川陕革命根
高新技术应用于农业的研究近来在国际上异常活跃,成果累累。1992年农业生物技术研究领域取得的重要新进展主要有以下10个方面: Recently, the application of high technol
AIM: To prepare neutralizing monoclonal antibody (mAb) against extracellular immunoglobulin (Ig)-like domainⅢ of vascular endothelial growth factor receptor KD
写在课前:rn为了加强教师之间的课堂教学交流,我们组织了一次区内教研联组的“同课异构”活动,以新课标小学数学实验教材二年级下册“1000以内数的认识”作为研究内容,由三位
小豆是一种营养丰富的杂粮作物且具有较高的药用和保健价值,但小豆产量低而不稳且品质差成为目前小豆发展的瓶颈问题。日照长度对小豆产量和籽粒品质的影响也是栽培领域值得研
Aim: To investigate the ultrastructural localization of atrial natriuretic peptide(ANP)-synthesizing cells and the relationship between ANP-synthesizing cells a