title: Evaluating ancient Egyptian prescriptions today: Anti-inflammatory activity of Ziziphus spina-christi.
abstract: BACKGROUND
  Ziziphus spina-christi (L.) Desf. (Christ's Thorn Jujube) is a wild tree today found in Jordan, Israel, Egypt, and some parts of Africa, which was already in use as a medicinal plant in Ancient Egypt. In ancient Egyptian prescriptions, it was used in remedies against swellings, pain, and heat, and thus should have anti-inflammatory effects. Nowadays, Z. spina-christi, is used in Egypt (by Bedouins, and Nubians), the Arabian Peninsula, Jordan, Iraq, and Morocco against a wide range of illnesses, most of them associated with inflammation. Pharmacological research undertaken to date suggests that it possesses anti-inflammatory, hypoglycemic, hypotensive and anti-microbial effects. The transcription factor NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) is critical in inflammation, proliferation and involved in various types of cancer. Identification of new anti-inflammatory compounds might be an effective strategy to target inflammatory disorders and cancer. Therefore, extracts from Z. spina-christi are investigated in terms of their anti-inflammatory effects. Our intention is to evaluate the effects of Z. spina-christi described in ancient Egyptian papyri, and to show whether the effects can be proven with modern pharmacological methods. Furthermore, we determine the active ingredients in crude extracts for their inhibitory activity toward NF-κB pathway.
  
  
  METHODS
  To determine the active ingredients of Z. spina-christi, we fractionated the extracts for bioassays and identified the active compounds. Epigallocatechin, gallocatechin, spinosin, 6''' feruloylspinosin and 6''' sinapoylspinosin and crude extracts of seed, leaf, root or stem were analyzed for their effect on NF-κB DNA binding by electromobility shift assay (EMSA) and nuclear translocation of NF-κB-p65 by Western blot analysis. The binding mode of the compounds to NF-κB pathway proteins was compared with the known inhibitor, MG-132, by in silico molecular docking calculations. Log10IC50 values of gallocatechin and epigallocatechin as two main compounds of the plant were correlated to the microarray-based mRNA expression of 79 inflammation-related genes in cell lines of the National Cancer Institute (NCI, USA) as determined. The expression of 17 genes significantly correlated to the log10IC50 values for gallocatechin or epigallocatechin.
  
  
  RESULTS
  Nuclear p65 protein level decreased upon treatment with each extract and compound. Root and seed extracts inhibited NF-κB-DNA binding as shown by EMSA. The compounds showed comparable binding energies and similar docking poses as MG-132 on the target proteins.
  
  
  CONCLUSIONS
  Z. spina-christi might possess anti-inflammatory activity as assumed by ancient Egyptian prescriptions. Five compounds contributed to this bioactivity, i.e. epigallocatechin, gallocatechin, spinosin, 6''' feruloylspinosin and 6''' sinapoylspinosin as shown in vitro and in silico.
authors.affiliation: Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Staudinger Weg 5, 55128 Mainz, Germany.
authors.forename: Onat
authors.initials: O
authors.lastname: Kadioglu
authors.affiliation: Institut für Biotechnologie und Wirkstoff Forschung gGmbH, Erwin-Schrödinger-Straße 56, 67663 Kaiserslautern, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Jacob
authors.affiliation: Institut für Biotechnologie und Wirkstoff Forschung gGmbH, Erwin-Schrödinger-Straße 56, 67663 Kaiserslautern, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Bohnert
authors.affiliation: Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Staudinger Weg 5, 55128 Mainz, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Naß
authors.affiliation: Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Staudinger Weg 5, 55128 Mainz, Germany.
authors.forename: Mohamed E M
authors.initials: ME
authors.lastname: Saeed
authors.affiliation: Department of Pharmacognosy, University of Khartoum, Khartoum, Sudan.
authors.forename: Hassan
authors.initials: H
authors.lastname: Khalid
authors.affiliation: Department of Pharmaceutical Biology and Biotechnology, Institute of Pharmaceutical Sciences, Albert-Ludwigs-University Freiburg, Stefan-Meier-Straße 19, 79104 Freiburg, Germany.
authors.forename: Irmgard
authors.initials: I
authors.lastname: Merfort
authors.affiliation: Institut für Biotechnologie und Wirkstoff Forschung gGmbH, Erwin-Schrödinger-Straße 56, 67663 Kaiserslautern, Germany; Institute of Biotechnology and Drug Research, Johannes Gutenberg University, Duesbergweg 10-14, 55128 Mainz, Germany.
authors.forename: Eckhard
authors.initials: E
authors.lastname: Thines
authors.affiliation: Department of Egyptology, Institute of Ancient Studies, Johannes Gutenberg University, Hegelstraße 59, 55122 Mainz, Germany. Electronic address: tpommere@uni-mainz.de.
authors.forename: Tanja
authors.initials: T
authors.lastname: Pommerening
authors.affiliation: Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Staudinger Weg 5, 55128 Mainz, Germany. Electronic address: efferth@uni-mainz.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Efferth
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D000076144:Egypt, Ancient; D029001:Herbal Medicine / Q000266:history; D049690:History, Ancient; D006801:Humans; D007249:Inflammation / Q000188:drug therapy; D007976:Leupeptins; D062105:Molecular Docking Simulation; D010936:Plant Extracts / Q000494:pharmacology*; D018517:Plant Roots / Q000737:chemistry; D018547:Plant Stems / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry; D051996:Transcription Factor RelA / Q000378:metabolism; D031957:Ziziphus / Q000737:chemistry*
chemical_list: D000893:Anti-Inflammatory Agents; D007976:Leupeptins; D010936:Plant Extracts; C498143:RELA protein, human; D051996:Transcription Factor RelA; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde
keywords: Ancient Egyptian medicinal papyri; Ethnopharmacology; Inflammation; Microarray; Molecular docking; Rhamnaceae