title: Acid sphingomyelinase as target of Lycium Chinense: promising new action for cell health.
abstract: BACKGROUND
  Sphingomyelin plays very important roles in cell function under physiological and pathological conditions. Physical and chemical stimuli produce reactive oxygen species that stimulate acid sphingomyelinase to induce apoptosis. Antioxidant plants of the traditional Chinese Pharmacopoeia, such as Lycium Barbarum and Lycium Chinense, have become increasingly popular in Western countries. We investigated the effects of Lycium Chinense on acid sphingomyelinase and sphingomyelin species in relation to gene expression.
  
  
  METHODS
  We prepared Lycium Chinense berry extracts and evaluated their antioxidant properties. Increasing amount of extracts was used to test cytotoxic and genotoxic effect on HepG2 cells. Gene expression, protein amount and enzyme activity of acid sphingomyelinase were tested by RT-PCR, immunoblotting and enzymatic activity assay, respectively. Sphingomyelin species were analyzed by UFLC MS/MS. A panel of 96 genes involved in oxidative stress, proliferation, apoptosis and cancer was used to test the effect of LC on gene expression. GLRX2, RNF7, and PTGS1 proteins were analyzed by immunoblotting.
  
  
  RESULTS
  We showed that Lycium Chinense berries have high antioxidant properties, have an IC50value of 9.55 mg/mL, do not induce genotoxic effect and maintain high level of cell viability. The berry extracts inhibit acid sphingomyelinase activity and increase both very long fatty acid sphingomyelin species and unsaturated fatty acid sphingomyelin species. Among 96 genes, Lycium Chinense berries up-regulate Glutaredoxin 2 and Ring Finger Protein 7 genes and proteins, able to protect cells from apoptosis. Intrigantly, Lycium Chinense berries down-regulates Prostaglandin H synthase 1 gene but the protein is not expressed in HepG2 cells.
  
  
  CONCLUSIONS
  The results identify acid sphingomyelinase as a novel target of Lycium Chinense berries to decrease saturated/unsaturated fatty acid sphingomyelin ratio, known to be useful for cell health. Consistent with these data, the berries regulate specifically gene expression to protect cells from apoptosis.
authors.affiliation: Department of Pharmaceutical Science, University of Perugia, Via Fabretti 48, 06122, Perugia, Italy.
authors.forename: Maria Rachele
authors.initials: MR
authors.lastname: Ceccarini
authors.affiliation: Department of Pharmaceutical Science, University of Perugia, Via Fabretti 48, 06122, Perugia, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Codini
authors.affiliation: Department of Pharmaceutical Science, University of Perugia, Via Fabretti 48, 06122, Perugia, Italy.
authors.forename: Samuela
authors.initials: S
authors.lastname: Cataldi
authors.affiliation: Department of Pharmaceutical Science, University of Perugia, Via Fabretti 48, 06122, Perugia, Italy.
authors.forename: Samuele
authors.initials: S
authors.lastname: Vannini
authors.affiliation: Laboratory of Nuclear Lipid BioPathology, CRABiON, Perugia, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Lazzarini
authors.affiliation: Laboratory of Nuclear Lipid BioPathology, CRABiON, Perugia, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Floridi
authors.affiliation: Department of Pharmaceutical Science, University of Perugia, Via Fabretti 48, 06122, Perugia, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Moretti
authors.affiliation: Department of Pharmaceutical Science, University of Perugia, Via Fabretti 48, 06122, Perugia, Italy.
authors.forename: Milena
authors.initials: M
authors.lastname: Villarini
authors.affiliation: Department of Chemistry, Biology and Biotechnology, University of Perugia, Via Elce di Sotto 8, 06123, Perugia, Italy.
authors.forename: Bernard
authors.initials: B
authors.lastname: Fioretti
authors.affiliation: Department of Pharmaceutical Science, University of Perugia, Via Fabretti 48, 06122, Perugia, Italy.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Beccari
authors.affiliation: Laboratory of Nuclear Lipid BioPathology, CRABiON, Perugia, Italy. elisabetta.albi@gmail.com.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Albi
mesh_terms: D000975:Antioxidants / Q000008:administration & dosage / Q000737:chemistry; D002470:Cell Survival / Q000187:drug effects*; D005638:Fruit / Q000737:chemistry; D005786:Gene Expression Regulation / Q000187:drug effects; D056945:Hep G2 Cells; D006801:Humans; D032305:Lycium / Q000737:chemistry; D008516:Medicine, Chinese Traditional; D010936:Plant Extracts / Q000008:administration & dosage / Q000737:chemistry; D013108:Sphingomyelin Phosphodiesterase / Q000096:biosynthesis*; D013109:Sphingomyelins / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D010936:Plant Extracts; D013109:Sphingomyelins; C112980:acid sphingomyelinase-1; D013108:Sphingomyelin Phosphodiesterase
keywords: Antioxidant; Fatty acids; Lycium chinense; Sphingomyelin; Sphingomyelinase