Enhanced anti-fibrogenic effects of novel oridonin derivative CYD0692 in hepatic stellate cells.
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 ~0.7 μM for 48 h, ~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 5 μM of CYD0692 treatment for 48 h, while 30 % inhibited at 10 μM. 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.
Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.|Shriners Hospital for Children-Galveston, 815 Market St., Galveston, TX, 77550, USA.
Fredrick J
FJ
Bohanon
Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.
Xiaofu
X
Wang
Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.|School of Medicine, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.
Brittany M
BM
Graham
Department of Cell Biology and Neuroscience, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.|Shriners Hospital for Children-Galveston, 815 Market St., Galveston, TX, 77550, USA.
Anesh
A
Prasai
Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.
Sadhashiva J
SJ
Vasudevan
Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.
Chunyong
C
Ding
Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.
Ye
Y
Ding
Department of Pediatrics, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.
Geetha L
GL
Radhakrishnan
Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA.
Cristiana
C
Rastellini
Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. jizhou@utmb.edu.
Jia
J
Zhou
Department of Surgery, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. rsradhak@utmb.edu.|Department of Pediatrics, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. rsradhak@utmb.edu.
Ravi S
RS
Radhakrishnan
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D045786:Diterpenes, Kaurane / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D055166:Hepatic Stellate Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D020128:Inhibitory Concentration 50; D008103:Liver Cirrhosis / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D051381:Rats; D059807:S Phase Cell Cycle Checkpoints / Q000187:drug effects; D013997:Time Factors
D051017:Apoptosis Regulatory Proteins; C000605270:CYD0682; D018797:Cell Cycle Proteins; D045786:Diterpenes, Kaurane; C011959:oridonin
Apoptosis; ECM; Liver fibrosis; Oridonin; Stellate cells