title: [Preparation of polyclonal antibody of recombinant human thioredoxin-1 and its protective effects on neonatal rats with endotoxemia].
abstract: OBJECTIVE
  To clone the gene human thioredoxin 1 (hTrx-1) expressing its protein in the E.coli expression system and to obtain its polyclonal antibody, and to study the protective effects of hTrx-1 on neonatal rats with endotoxemia.
  
  
  METHODS
  DNA encoding hTrx-1 from fetal liver cells was isolated by RT-PCR. The hTrx-1 was cloned to the prokaryotic expression plasmid PET-28a to induce its protein expression in the E.coli expression system. The purified hTrx-1 was injected into rats to prepare polyclonal antibody. Newborn Sprague-Dawley rats were randomly assigned to three groups: control, lipopolysaccharide (LPS) and hTrx-1 (n=12 each). The control and the LPS groups were intraperitoneally injected with normal saline and LPS (5 mg/kg), respectively. The hTrx-1 group received an intraperitoneal injection of hTrx-1 (10 mg/kg) 30 minutes before LPS injection. The mortality rate 24 hrs after injection was compared between the three groups.
  
  
  RESULTS
  The prokaryotic expression plasmid PET-28a-hTrx-1 was constructed. The hTrx-1 protein was expressed and purified. The polyclonal antibody of hTrx-1 with the titer of 1∶51200 was prepared. The mortality rate of the control, LPS and hTrx-1 groups was 0, 67% and 17%, respectively (χ2=14.400, P<0.01).
  
  
  CONCLUSIONS
  The polyclonal antibody of hTrx-1 is prepared successfully. The hTrx-1 protein has protective effects on neonatal rats with endotoxiamia.
authors: Department of Neonatology, Nanfang Hospital of Southern Medical University, Guangzhou, China.
authors: Hai-Yan
authors: HY
authors: Zhao
authors: Hong-Wu
authors: HW
authors: Chen
authors: Xin-Hua
authors: XH
authors: Qian
authors: Xu-Chu
authors: XC
authors: Hu
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000906:Antibodies / Q000032:analysis*; D015153:Blotting, Western; D019446:Endotoxemia / Q000517:prevention & control*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000276:immunology; D013879:Thioredoxins / Q000235:genetics / Q000276:immunology* / Q000627:therapeutic use*
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins; C491203:TXN protein, human; D013879:Thioredoxins