Enhanced anti-fibrogenic effects of novel oridonin derivative CYD0692 in hepatic stellate cells

Fredrick J. Bohanon, Xiaofu Wang, Brittany M. Graham, Anesh Prasai, Sadhashiva J. Vasudevan, Chunyong Ding, Ye Ding, Geetha Radhakrishnan, Cristiana Rastellini, Jia Zhou, Ravi Radhakrishnan

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)293-300
Number of pages8
JournalMolecular and Cellular Biochemistry
Volume410
Issue number1-2
DOIs
StatePublished - Sep 7 2015

Fingerprint

Hepatic Stellate Cells
Transforming Growth Factors
Cells
Derivatives
Fibronectins
Isodon
Apoptosis
Cyclin B1
Military electronic countermeasures
Flow cytometry
Liver
Cell proliferation
Cell growth
Cell death
Collagen Type I
Cell Cycle Checkpoints
S Phase
Inhibitory Concentration 50
Smooth Muscle
Muscle

Keywords

  • Apoptosis
  • ECM
  • Liver fibrosis
  • Oridonin
  • Stellate cells

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Enhanced anti-fibrogenic effects of novel oridonin derivative CYD0692 in hepatic stellate cells. / Bohanon, Fredrick J.; Wang, Xiaofu; Graham, Brittany M.; Prasai, Anesh; Vasudevan, Sadhashiva J.; Ding, Chunyong; Ding, Ye; Radhakrishnan, Geetha; Rastellini, Cristiana; Zhou, Jia; Radhakrishnan, Ravi.

In: Molecular and Cellular Biochemistry, Vol. 410, No. 1-2, 07.09.2015, p. 293-300.

Research output: Contribution to journalArticle

Bohanon, Fredrick J. ; Wang, Xiaofu ; Graham, Brittany M. ; Prasai, Anesh ; Vasudevan, Sadhashiva J. ; Ding, Chunyong ; Ding, Ye ; Radhakrishnan, Geetha ; Rastellini, Cristiana ; Zhou, Jia ; Radhakrishnan, Ravi. / Enhanced anti-fibrogenic effects of novel oridonin derivative CYD0692 in hepatic stellate cells. In: Molecular and Cellular Biochemistry. 2015 ; Vol. 410, No. 1-2. pp. 293-300.
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AU - Prasai, Anesh

AU - Vasudevan, Sadhashiva J.

AU - Ding, Chunyong

AU - Ding, Ye

AU - Radhakrishnan, Geetha

AU - Rastellini, Cristiana

AU - Zhou, Jia

AU - Radhakrishnan, Ravi

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AB - 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.

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