title: [Euphorbia fischeriana extract reactivates latent HIV through nuclear factor-κB pathway].
abstract: OBJECTIVE
  To investigate the effect of Euphorbia fischeriana extract on latent HIV reactivation and the pathway involved in this process and discuss the value of Euphorbia fischeriana extract in eliminating HIV.
  
  
  METHODS
  Fresh tissues of Euphorbia fischeriana root were crushed into powder after quick freezing with liquid nitrogen and extracted with acetone followed by a three-day vacuum freeze-drying for dehydration of the extract. The extract (EFE) was separated using RP-C18 column with high-performance liquid chromatography (HPLC) and identified with mass spectrometry (MS). The activity of reactivated latent HIV was analyzed by fluorescence-activated cell sorting in a J-Lat 10.6 cell model treated with EFE (50 µg/mL) for 24 h, using TNF-α (10 ng/mL) as the positive control. The effect of a NF-κB pathway inhibitor (Bay 11-7082) on EFE activity was tested. The changes in P65 expression in the cell nuclei within 2 h and HIV protein p24 expression within 24 h were analyzed by Western blotting in cells treated with EFE.
  
  
  RESULTS
  EFE was obtained by one-step acetone extraction, and the concentration of prostratin in the extract was around 0.53 mmol/L. About 50% of the cells showed HIV reactivation after treatment with 50 µg/mL EFE for 24 h accompanied by a significantly increased p24 expression. The activity of EFE in reactivating latent HIV was inhibited by Bay 11-7082 in a concentration-dependent manner, and p65 accumulation was detected in the cell nuclei within 2 h.
  
  
  CONCLUSIONS
  EFE we obtained contains the active compounds of prostratin and its analogues and shows a strong capacity to reactivate latent HIV through classical NF-κB pathway.
authors.affiliation: School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China.E-mail: xiaoyanpan1201@163.com.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Pan
authors.forename: Mingjiao
authors.initials: M
authors.lastname: Zhang
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Zeng
authors.forename: Jian
authors.initials: J
authors.lastname: Lin
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.forename: Minmin
authors.initials: M
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
mesh_terms: D028481:Euphorbia / Q000737:chemistry*; D005434:Flow Cytometry; D006678:HIV / Q000187:drug effects*; D015658:HIV Infections; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D009570:Nitriles; D010703:Phorbol Esters / Q000737:chemistry; D010936:Plant Extracts / Q000494:pharmacology*; D015398:Signal Transduction; D013450:Sulfones; D014409:Tumor Necrosis Factor-alpha; D017735:Virus Latency / Q000187:drug effects*
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D016328:NF-kappa B; D009570:Nitriles; D010703:Phorbol Esters; D010936:Plant Extracts; D013450:Sulfones; C439524:TNF protein, human; D014409:Tumor Necrosis Factor-alpha; C070999:prostratin