title: Changes in amount of ADM mRNA and RAMP2 mRNA in calcified vascular smooth muscle cells.
abstract: This work was aimed to explore the changes and significance of adrenomedullin (ADM) mRNA and receptor activity modifying protein 2 (RAMP2) mRNA in calcified vascular smooth muscle cells (VSMCs). Calcification of cultured rat VSMCs was produced by incubation with beta-glycerophosphate. Content of ADM released by VSMCs was measured by radioimmunoassay (RIA). The amount of ADM mRNA and RAMP2 mRNA was determined by competitive quantitative RT-PCR. The intracellular calcium content, alkaline phosphatases activity and cellular (45)Ca(2+)-uptake were determined. The results showed that the content of calcium, (45)Ca(2+)-uptake and alkaline phosphatases activity in calcified VSMCs were increased by 118%, 174% and seven-fold (all P<0.01), respectively, compared with control VSMCs. Content of ADM in medium was increased by 99% (P<0.01). Furthermore, it was found that the amount of ADM mRNA and RAMP2 mRNA in calcified cells was elevated by 78 and 56% (all P<0.05), respectively, compared with control. The elevated levels of RAMP2 mRNA were in positive correlation with ADM mRNA (r=0.76, P<0.05) in calcified VSMCs. In conclusion, calcified VSMCs generated an increased amount of ADM, and up-regulated gene expressions of ADM and RAMP2.
authors.affiliation: Institute of Cardiovascular Diseases, Peking University First Hospital, 100034, PR, Beijing, China.
authors.forename: Yong Fen
authors.initials: YF
authors.lastname: Qi
authors.forename: Shu Heng
authors.initials: SH
authors.lastname: Wang
authors.forename: Bao Hong
authors.initials: BH
authors.lastname: Zhang
authors.forename: Ding Fang
authors.initials: DF
authors.lastname: Bu
authors.forename: Tang Chao
authors.initials: TC
authors.lastname: Shu
authors.forename: Jun Bao
authors.initials: JB
authors.lastname: Du
mesh_terms: D053607:Adrenomedullin; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002122:Calcium Chloride / Q000494:pharmacology; D002478:Cells, Cultured; D005994:Glycerophosphates / Q000737:chemistry; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000235:genetics*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D010455:Peptides / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats; D017208:Rats, Wistar; D058263:Receptor Activity-Modifying Protein 2; D058261:Receptor Activity-Modifying Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
chemical_list: D005994:Glycerophosphates; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D010455:Peptides; D012333:RNA, Messenger; C545509:Ramp2 protein, rat; D058263:Receptor Activity-Modifying Protein 2; D058261:Receptor Activity-Modifying Proteins; D053607:Adrenomedullin; D000469:Alkaline Phosphatase; D002122:Calcium Chloride; D002118:Calcium; C031463:beta-glycerophosphoric acid