title: Enhanced effects of novel oridonin analog CYD0682 for hepatic fibrosis.
abstract: BACKGROUND
  Activated hepatic stellate cells (HSCs) are responsible for excess extracellular matrix (ECM) protein deposition in liver fibrosis. Previously, our group reported that the natural compound oridonin induces apoptosis, inhibits cell proliferation, and downregulates ECM proteins in activated HSC. In this study, the antifibrogenic effects of oridonin derivative CYD0682 on the activated human LX-2 and rat HSC-T6 stellate cell lines were investigated.
  
  
  METHODS
  Cell proliferation was measured by alamarBlue assay. Apoptosis was detected by Cell Death ELISA and staining of Yo-Pro-1 and propidium iodide. Cell cycle was determined by flow cytometry. Immunoblot and immunofluorescence staining were performed for cellular protein expression.
  
  
  RESULTS
  CYD0682 treatment significantly inhibited LX-2 cell proliferation in a dose- and time-dependent manner with an IC50 value of 0.49 μM for 48 h, ∼10-fold greater potency than oridonin. Similar results were observed in HSC-T6 cells. In contrast, 2.5 μM of CYD0682 treatment had no significant effects on proliferation of the human hepatocyte cell line C3A. CYD0682 treatment induced LX-2 cell apoptosis and S-phase cell cycle arrest and was associated with activation of p53, p21, and cleaved caspase-3. The myofibroblast marker protein α-smooth muscle actin and major ECM proteins type I collagen and fibronectin were markedly suppressed in a time- and dose-dependent fashion by CYD0682. Furthermore, pretreatment with CYD0682 blocked transforming growth factor-β-induced type I collagen and fibronectin production.
  
  
  CONCLUSIONS
  In comparison with oridonin, its novel derivative CYD0682 may act as a more potent antihepatic fibrosis agent.
authors.affiliation: Department of Surgery, The University of Texas Medical Branch, Galveston, Texas.
authors.forename: Fredrick J
authors.initials: FJ
authors.lastname: Bohanon
authors.affiliation: Department of Surgery, The University of Texas Medical Branch, Galveston, Texas.
authors.forename: Xiaofu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Surgery, The University of Texas Medical Branch, Galveston, Texas.
authors.forename: Brittany M
authors.initials: BM
authors.lastname: Graham
authors.affiliation: Chemical Biology Program, Department of Pharmacology and Toxicology, The University of Texas Medical Branch, Galveston, Texas.
authors.forename: Chunyong
authors.initials: C
authors.lastname: Ding
authors.affiliation: Chemical Biology Program, Department of Pharmacology and Toxicology, The University of Texas Medical Branch, Galveston, Texas.
authors.forename: Ye
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Department of Pediatrics, The University of Texas Medical Branch, Galveston, Texas.
authors.forename: Geetha L
authors.initials: GL
authors.lastname: Radhakrishnan
authors.affiliation: Department of Surgery, The University of Texas Medical Branch, Galveston, Texas.
authors.forename: Cristiana
authors.initials: C
authors.lastname: Rastellini
authors.affiliation: Chemical Biology Program, Department of Pharmacology and Toxicology, The University of Texas Medical Branch, Galveston, Texas. Electronic address: jizhou@utmb.edu.
authors.forename: Jia
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Department of Surgery, The University of Texas Medical Branch, Galveston, Texas; Department of Pediatrics, The University of Texas Medical Branch, Galveston, Texas. Electronic address: rsradhak@utmb.edu.
authors.forename: Ravi S
authors.initials: RS
authors.lastname: Radhakrishnan
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D045786:Diterpenes, Kaurane / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use*; D015536:Down-Regulation / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D055166:Hepatic Stellate Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008103:Liver Cirrhosis / Q000188:drug therapy*; D051381:Rats; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: C000605270:CYD0682; D045786:Diterpenes, Kaurane; D016326:Extracellular Matrix Proteins; D016212:Transforming Growth Factor beta; C011959:oridonin
keywords: Derivatives; Fibrosis; Liver; Oridonin; Stellate cells