title: Establishing a low-expression renalase gene model in cardiac tissue of Sprague-Dawley rats.
abstract: BACKGROUND
  Renalase is a novel secretory amino oxidase expressed in the kidney and heart. To study the protective mechanism of renalase in local heart tissue, we established a low-expression renalase model with lentivirus (LV)-mediated RNA interference technology.
  
  
  METHODS
  Three renalase-targeting oligonucleotides were designed after analyzing the mRNA of renalase. LV particles were prepared with LV expression systems (using the Trono 3 plasmid component system), after which LV-RNLS-shRNAs and LV-NC-shRNA were transfected into H9C2 cells in different cell culture plates. The optimal oligonucleotide was screened by real-time PCR and Western blot. These techniques were also used to detect renalase gene expression in the heart tissue.
  
  
  RESULTS
  In the cell screening experiment, the efficacy of the inhibition of renalase mRNA expression was 93.7 % and that of renalase protein expression was 83.1 % in H9C2 cells. When the oligonucleotide was injected into the pericardial cavities of the SD rats on the 10th day, it inhibited 63.9 % of the expression of renalase protein in the heart tissue.
  
  
  CONCLUSIONS
  LV-RNLS-RNAi (19813-1) can be used to establish an optimal renalase low-expression model for further research on the renalase system.
authors.affiliation: Department of Cardiology, The Third Xiangya Hospital of Central South University, 410013, Changsha, Hunan, China.
authors.forename: X
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Cardiology, The Third Xiangya Hospital of Central South University, 410013, Changsha, Hunan, China.
authors.forename: M
authors.initials: M
authors.lastname: Lin
authors.affiliation: Department of Cardiology, The Third Xiangya Hospital of Central South University, 410013, Changsha, Hunan, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Xie
authors.affiliation: Department of Cardiology, The Third Xiangya Hospital of Central South University, 410013, Changsha, Hunan, China.
authors.forename: R
authors.initials: R
authors.lastname: Huang
authors.affiliation: Department of Cardiology, The Third Xiangya Hospital of Central South University, 410013, Changsha, Hunan, China. afychen@yahoo.com.
authors.forename: A F
authors.initials: AF
authors.lastname: Chen
authors.affiliation: Department of Cardiology, The Third Xiangya Hospital of Central South University, 410013, Changsha, Hunan, China. jiangweihongdoc@163.com.
authors.forename: W
authors.initials: W
authors.lastname: Jiang
mesh_terms: D000818:Animals; D015536:Down-Regulation / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic; D055785:Gene Knockdown Techniques / Q000379:methods*; D016086:Lentivirus / Q000235:genetics*; D008995:Monoamine Oxidase / Q000235:genetics / Q000378:metabolism*; D009206:Myocardium / Q000201:enzymology*; D034622:RNA Interference / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D055647:Rats, Transgenic; D014162:Transfection / Q000379:methods
chemical_list: D008995:Monoamine Oxidase; C521557:renalase
keywords: Heart; Kidney; Lentivirus; RNA interference; Renalase