title: Antiangiogenic effects of oridonin.
abstract: BACKGROUND
  Oridonin, the major terpene found in Rabdosia rubescens (Henmsl.) Hara, is widely used as a dietary supplement and therapeutic drug. Oridonin has been proven to possess good anti-tumour activity, but little is known about its effect on angiogenesis. The aim of this study was to investigate the antiangiogenic effects of oridonin in vivo and in vitro and prove that oridonin anti-tumour activity is based on suppressing angiogenesis.
  
  
  METHODS
  In vitro, the antiangiogenesis effect was studied by proliferation, apoptosis, migration, invasion, and tube formation experiments on human umbilical vascular endothelial cells (HUVECs). In vivo, using the Tg (fli1: GFP) zebrafish model, the embryonic vasculogenesis and postnatal regeneration were evaluated. The vascular endothelial growth factor (VEGF) signalling pathway gene expressions were assessed by reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, the inhibition effects on tumour growth and metastasis were observed using a xenograft zebrafish tumour model and xenograft nude mouse tumour model. Angiogenesis was assayed by immunostaining with cluster of differentiation 31. Importantly, the proteins were identified as being differentially expressed in an in vivo model by two-dimensional electrophoresis-mass spectrometry (2D-MS) and western blot (WB).
  
  
  RESULTS
  The results indicated that oridonin inhibited HUVEC proliferation, migration, invasion, and tube formation and induced cell apoptosis. Oridonin inhibited zebrafish angiogenesis during embryonic development and tail fin regeneration. RT-PCR showed that oridonin decreased the VEGFA, VEGFR2, and VEGFR3 expressions in zebrafish, while the TP53 expression increased. Moreover, oridonin had strong effects on tumour growth and metastasis in vivo. 2D-MS identified a total of 50 proteins differentially expressed (17 up-expressed, 28 down-expressed). Lastly, WB showed that Claudin 1, Claudin 4, and Claudin 7 were closely related to tumour growth and metastasis.
  
  
  CONCLUSIONS
  This study demonstrated that oridonin could inhibit tumour growth and metastasis, which mainly based on oridonin antiangiogenic effects. Claudin 1, Claudin 4, and Claudin 7 were the main contributors to the mechanism.
authors.affiliation: Traditional Chinese medicine pharmacy, Zhejiang Hospital, No. 12 Lingyin Road, Xihu District, Zhejiang, Hangzhou, China.|School of Pharmacy, Shanghai University of Traditional Chinese Medicine, No. 1200 Cailun Road, Pudong New District, Shanghai, 201203, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Tian
authors.affiliation: Anesthesia Department, Zhejiang Cancer Hospital, No. 38 Guangji Road, Gongshu District, Zhejiang, Hangzhou, 310002, China.
authors.forename: Kangjie
authors.initials: K
authors.lastname: Xie
authors.affiliation: Institute of Developmental and Regenerative Biology, Hangzhou Normal University, No. 16 Xuelin Street, Jianggan District, Zhejiang, Hangzhou, 310036, China.
authors.forename: Donglai
authors.initials: D
authors.lastname: Sheng
authors.affiliation: Traditional Chinese medicine pharmacy, Zhejiang Hospital, No. 12 Lingyin Road, Xihu District, Zhejiang, Hangzhou, China.
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Wan
authors.affiliation: School of Pharmacy, Shanghai University of Traditional Chinese Medicine, No. 1200 Cailun Road, Pudong New District, Shanghai, 201203, China. zgfzyy2015@sina.com.
authors.forename: Guofu
authors.initials: G
authors.lastname: Zhu
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology; D045786:Diterpenes, Kaurane / Q000494:pharmacology*; D004354:Drug Screening Assays, Antitumor; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D037602:Isodon / Q000737:chemistry*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D015027:Zebrafish
chemical_list: D020533:Angiogenesis Inhibitors; D000972:Antineoplastic Agents, Phytogenic; D045786:Diterpenes, Kaurane; C011959:oridonin
keywords: Angiogenesis; HUVECs; Mouse; Oridonin; Zebrafish