caa a22 4500 606235841 CHVBK 20210128101244.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s11010-015-2562-4 doi (NATIONALLICENCE)springer-10.1007/s11010-015-2562-4 Enhanced anti-fibrogenic effects of novel oridonin derivative CYD0692 in hepatic stellate cells [Elektronische Daten] [Fredrick Bohanon, Xiaofu Wang, Brittany Graham, Anesh Prasai, Sadhashiva Vasudevan, Chunyong Ding, Ye Ding, Geetha Radhakrishnan, Cristiana Rastellini, Jia Zhou, Ravi Radhakrishnan] Oridonin, isolated from Rabdosia rubescens, has been proven to possess various anti-neoplastic and anti-inflammatory properties. Previously, we reported the anti-fibrogenic effects of oridonin for liver in vitro. In the present study, we investigated the effects of a newly designed analog CYD0692 in vitro. Cell viability was measured by Alamar Blue assay. Cell apoptosis was assessed by Cell Death ELISA and Yo-Pro-1 staining. Western blots were performed for cellular proteins. Flow cytometry was used to measure cell cycle regulation. CYD0692 significantly inhibited LX-2 cells proliferation in a dose- and time-dependent manner with an IC50 value of ~0.7μM for 48h, ~tenfold greater potency than oridonin. Similar results were observed in HSC-T6 cells. In contrast, on the human hepatocyte cell line C3A, only 12% of the cell growth was inhibited with 5μM of CYD0692 treatment for 48h, while 30% inhibited at 10μM. After CYD0692 treatment on LX-2 cells, apoptosis and S-phase cell cycle arrest were induced; cleaved-PARP, p21, and p53 were activated while cyclin-B1 levels declined. In addition, α-smooth muscle actin, type I Collagen, and fibronectin (FN) were markedly down regulated. Transforming growth factor β1 (TGF β1) has been identified as a dominant stimulator for ECM production in HSC. Our results indicated that pretreatment with CYD0692 blocked TGF β1-induced FN expression, thereby decreasing the downstream factors of TGF β1 signaling, such as Phospho-Smad2/3 and phospho-ERK. In comparison with oridonin, its novel derivative CYD0692 has demonstrated to be a more potent and potentially safer anti-fibrogenic agent for the treatment of hepatic fibrosis. Springer Science+Business Media New York, 2015 Oridonin nationallicence Liver fibrosis nationallicence Stellate cells nationallicence Apoptosis nationallicence ECM nationallicence Bohanon Fredrick Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Wang Xiaofu Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Graham Brittany Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Prasai Anesh Department of Cell Biology and Neuroscience, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Vasudevan Sadhashiva Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Ding Chunyong Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Ding Ye Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Radhakrishnan Geetha Department of Pediatrics, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Rastellini Cristiana Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Zhou Jia Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Radhakrishnan Ravi Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 410/1-2(2015-12-01), 293-300 0300-8177 410:1-2<293 2015 410 11010 https://doi.org/10.1007/s11010-015-2562-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-015-2562-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Bohanon Fredrick Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Wang Xiaofu Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Graham Brittany Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Prasai Anesh Department of Cell Biology and Neuroscience, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Vasudevan Sadhashiva Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Ding Chunyong Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Ding Ye Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Radhakrishnan Geetha Department of Pediatrics, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Rastellini Cristiana Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Zhou Jia Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 700 1- Radhakrishnan Ravi Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, 77555, Galveston, TX, USA aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 410/1-2(2015-12-01), 293-300 0300-8177 410:1-2<293 2015 410 11010