title: Protective effect of guggulsterone against cardiomyocyte injury induced by doxorubicin in vitro.
abstract: BACKGROUND
  Doxorubicin (DOX) is an effective antineoplastic drug; however, clinical use of DOX is limited by its dose-dependent cardiotoxicity. It is well known that reactive oxygen species (ROS) play a vital role in the pathological process of DOX-induced cardiotoxicity. For this study, we evaluated the protective effects of guggulsterone (GS), a steroid obtained from myrrh, to determine its preliminary mechanisms in defending against DOX-induced cytotoxicity in H9C2 cells.
  
  
  METHODS
  In this study, we used a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release measurements, and Hoechst 33258 staining to evaluate the protective effect of GS against DOX-induced cytotoxicity in H9C2 cells. In addition, we observed the immunofluorescence of intracellular ROS and measured lipid peroxidation, caspase-3 activity, and apoptosis-related proteins by using Western blotting.
  
  
  RESULTS
  The MTT assay and LDH release showed that treatment using GS (1-30 μM) did not cause cytotoxicity. Furthermore, GS inhibited DOX (1 μM)-induced cytotoxicity in a concentration-dependent manner. Hoechst 33258 staining showed that GS significantly reduced DOX-induced apoptosis and cell death. Using GS at a dose of 10-30 μM significantly reduced intracellular ROS and the formation of MDA in the supernatant of DOX-treated H9C2 cells and suppressed caspase-3 activity to reference levels. In immunoblot analysis, pretreatment using GS significantly reversed DOX-induced decrease of PARP, caspase-3 and bcl-2, and increase of bax, cytochrome C release, cleaved-PARP and cleaved-caspase-3. In addition, the properties of DOX-induced cancer cell (DLD-1 cells) death did not interfere when combined GS and DOX.
  
  
  CONCLUSIONS
  These data provide considerable evidence that GS could serve as a novel cardioprotective agent against DOX-induced cardiotoxicity.
authors.affiliation: Department of Surgery, Chi Mei Medical Center, Tainan, Taiwan.
authors.forename: Wen-Ching
authors.initials: WC
authors.lastname: Wang
authors.forename: Yih-Huei
authors.initials: YH
authors.lastname: Uen
authors.forename: Ming-Long
authors.initials: ML
authors.lastname: Chang
authors.forename: Khoot-Peng
authors.initials: KP
authors.lastname: Cheah
authors.forename: Joe-Sharg
authors.initials: JS
authors.lastname: Li
authors.forename: Wen-Yu
authors.initials: WY
authors.lastname: Yu
authors.forename: Kock-Chee
authors.initials: KC
authors.lastname: Lee
authors.forename: Cheuk-Sing
authors.initials: CS
authors.lastname: Choy
authors.forename: Chien-Ming
authors.initials: CM
authors.lastname: Hu
mesh_terms: D000970:Antineoplastic Agents / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004317:Doxorubicin / Q000633:toxicity*; D006801:Humans; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011282:Pregnenediones / Q000494:pharmacology*; D020011:Protective Agents / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013729:Terpenes / Q000737:chemistry*
chemical_list: D000970:Antineoplastic Agents; D011282:Pregnenediones; D020011:Protective Agents; D017382:Reactive Oxygen Species; D013729:Terpenes; D004317:Doxorubicin; C023617:pregna-4,17-diene-3,16-dione; C014119:myrrh oil