title: Effects of endogenous substance P expression on degranulation in RBL-2H3 cells.
abstract: OBJECTIVE
  To determine whether RBL-2H3 cells have endogenous substance P (SP) expression under immunoglobulin E (IgE)-activated and inactivated conditions, and to ascertain the function of endogenous SP in the antigen-induced degranulation of RBL-2H3 cells.
  
  
  METHODS
  SP mRNA and protein expression in both inactivated and 2,4-dinitrophenol (DNP)-specific IgE-activated RBL-2H3 cells were assessed by real-time PCR and immunofluorescence, respectively. Following activation with DNP-specific IgE, the degranulation of RBL-2H3 cells in response to DNP-bovine serum albumin (BSA), with and without endogenous SP expression, was assessed by monitoring the release of the granular enzyme β-hexosaminidase.
  
  
  RESULTS
  Endogenous SP mRNA and peptide expression increased in activated RBL-2H3 cells, compared with inactivated RBL-2H3 cells. The small hairpin RNA (shRNA)-mediated knockdown of endogenous SP reduced the degranulation ability of RBL-2H3 cells.
  
  
  CONCLUSIONS
  Activated RBL-2H3 cells express endogenous SP which plays a role in antigen-induced degranulation.
authors.affiliation: Department of ENT, Huadong Hospital, Shanghai Medical School, Fudan University, Shanghai, 200032, People's Republic of China.
authors.forename: Hua
authors.initials: H
authors.lastname: Hu
authors.forename: Ruxin
authors.initials: R
authors.lastname: Zhang
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Fang
authors.forename: Min
authors.initials: M
authors.lastname: Yu
authors.forename: Shaoqing
authors.initials: S
authors.lastname: Yu
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
mesh_terms: D000818:Animals; D015550:Cell Degranulation / Q000187:drug effects; D045744:Cell Line, Tumor; D002540:Cerebral Cortex / Q000276:immunology; D004140:Dinitrophenols / Q000494:pharmacology; D055785:Gene Knockdown Techniques; D007073:Immunoglobulin E / Q000276:immunology; D008407:Mast Cells / Q000187:drug effects / Q000276:immunology*; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013373:Substance P / Q000235:genetics / Q000276:immunology*
chemical_list: D004140:Dinitrophenols; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C034638:dinitrophenyl-bovine serum albumin; D012710:Serum Albumin, Bovine; D013373:Substance P; D007073:Immunoglobulin E