Enhanced effects of novel oridonin analog CYD0682 for hepatic fibrosis

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

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Background Activated hepatic stellate cells (HSCs) are responsible for excess extracellular matrix (ECM) protein deposition in liver fibrosis. Previously, our group reported that the natural compound oridonin induces apoptosis, inhibits cell proliferation, and downregulates ECM proteins in activated HSC. In this study, the antifibrogenic effects of oridonin derivative CYD0682 on the activated human LX-2 and rat HSC-T6 stellate cell lines were investigated. Methods Cell proliferation was measured by alamarBlue assay. Apoptosis was detected by Cell Death ELISA and staining of Yo-Pro-1 and propidium iodide. Cell cycle was determined by flow cytometry. Immunoblot and immunofluorescence staining were performed for cellular protein expression. Results CYD0682 treatment significantly inhibited LX-2 cell proliferation in a dose- and time-dependent manner with an IC50 value of 0.49 μM for 48 h, ∼10-fold greater potency than oridonin. Similar results were observed in HSC-T6 cells. In contrast, 2.5 μM of CYD0682 treatment had no significant effects on proliferation of the human hepatocyte cell line C3A. CYD0682 treatment induced LX-2 cell apoptosis and S-phase cell cycle arrest and was associated with activation of p53, p21, and cleaved caspase-3. The myofibroblast marker protein α-smooth muscle actin and major ECM proteins type I collagen and fibronectin were markedly suppressed in a time- and dose-dependent fashion by CYD0682. Furthermore, pretreatment with CYD0682 blocked transforming growth factor-β–induced type I collagen and fibronectin production. Conclusions In comparison with oridonin, its novel derivative CYD0682 may act as a more potent antihepatic fibrosis agent.

Original languageEnglish (US)
Pages (from-to)441-449
Number of pages9
JournalJournal of Surgical Research
Volume199
Issue number2
DOIs
StatePublished - Dec 1 2015

Fingerprint

Fibrosis
Hepatic Stellate Cells
Liver
Extracellular Matrix Proteins
Cell Proliferation
Apoptosis
Collagen Type I
Fibronectins
Staining and Labeling
Cell Line
Myofibroblasts
Propidium
Transforming Growth Factors
Cell Cycle Checkpoints
oridonin
CYD0682
S Phase
Caspase 3
Liver Cirrhosis
Inhibitory Concentration 50

Keywords

  • Derivatives
  • Fibrosis
  • Liver
  • Oridonin
  • Stellate cells

ASJC Scopus subject areas

  • Surgery
  • Medicine(all)

Cite this

Enhanced effects of novel oridonin analog CYD0682 for hepatic fibrosis. / Bohanon, Fredrick J.; Wang, Xiaofu; Graham, Brittany M.; Ding, Chunyong; Ding, Ye; Radhakrishnan, Geetha; Rastellini, Cristiana; Zhou, Jia; Radhakrishnan, Ravi.

In: Journal of Surgical Research, Vol. 199, No. 2, 01.12.2015, p. 441-449.

Research output: Contribution to journalArticle

Bohanon, Fredrick J. ; Wang, Xiaofu ; Graham, Brittany M. ; Ding, Chunyong ; Ding, Ye ; Radhakrishnan, Geetha ; Rastellini, Cristiana ; Zhou, Jia ; Radhakrishnan, Ravi. / Enhanced effects of novel oridonin analog CYD0682 for hepatic fibrosis. In: Journal of Surgical Research. 2015 ; Vol. 199, No. 2. pp. 441-449.
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abstract = "Background Activated hepatic stellate cells (HSCs) are responsible for excess extracellular matrix (ECM) protein deposition in liver fibrosis. Previously, our group reported that the natural compound oridonin induces apoptosis, inhibits cell proliferation, and downregulates ECM proteins in activated HSC. In this study, the antifibrogenic effects of oridonin derivative CYD0682 on the activated human LX-2 and rat HSC-T6 stellate cell lines were investigated. Methods Cell proliferation was measured by alamarBlue assay. Apoptosis was detected by Cell Death ELISA and staining of Yo-Pro-1 and propidium iodide. Cell cycle was determined by flow cytometry. Immunoblot and immunofluorescence staining were performed for cellular protein expression. Results CYD0682 treatment significantly inhibited LX-2 cell proliferation in a dose- and time-dependent manner with an IC50 value of 0.49 μM for 48 h, ∼10-fold greater potency than oridonin. Similar results were observed in HSC-T6 cells. In contrast, 2.5 μM of CYD0682 treatment had no significant effects on proliferation of the human hepatocyte cell line C3A. CYD0682 treatment induced LX-2 cell apoptosis and S-phase cell cycle arrest and was associated with activation of p53, p21, and cleaved caspase-3. The myofibroblast marker protein α-smooth muscle actin and major ECM proteins type I collagen and fibronectin were markedly suppressed in a time- and dose-dependent fashion by CYD0682. Furthermore, pretreatment with CYD0682 blocked transforming growth factor-β–induced type I collagen and fibronectin production. Conclusions In comparison with oridonin, its novel derivative CYD0682 may act as a more potent antihepatic fibrosis agent.",
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T1 - Enhanced effects of novel oridonin analog CYD0682 for hepatic fibrosis

AU - Bohanon, Fredrick J.

AU - Wang, Xiaofu

AU - Graham, Brittany M.

AU - Ding, Chunyong

AU - Ding, Ye

AU - Radhakrishnan, Geetha

AU - Rastellini, Cristiana

AU - Zhou, Jia

AU - Radhakrishnan, Ravi

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N2 - Background Activated hepatic stellate cells (HSCs) are responsible for excess extracellular matrix (ECM) protein deposition in liver fibrosis. Previously, our group reported that the natural compound oridonin induces apoptosis, inhibits cell proliferation, and downregulates ECM proteins in activated HSC. In this study, the antifibrogenic effects of oridonin derivative CYD0682 on the activated human LX-2 and rat HSC-T6 stellate cell lines were investigated. Methods Cell proliferation was measured by alamarBlue assay. Apoptosis was detected by Cell Death ELISA and staining of Yo-Pro-1 and propidium iodide. Cell cycle was determined by flow cytometry. Immunoblot and immunofluorescence staining were performed for cellular protein expression. Results CYD0682 treatment significantly inhibited LX-2 cell proliferation in a dose- and time-dependent manner with an IC50 value of 0.49 μM for 48 h, ∼10-fold greater potency than oridonin. Similar results were observed in HSC-T6 cells. In contrast, 2.5 μM of CYD0682 treatment had no significant effects on proliferation of the human hepatocyte cell line C3A. CYD0682 treatment induced LX-2 cell apoptosis and S-phase cell cycle arrest and was associated with activation of p53, p21, and cleaved caspase-3. The myofibroblast marker protein α-smooth muscle actin and major ECM proteins type I collagen and fibronectin were markedly suppressed in a time- and dose-dependent fashion by CYD0682. Furthermore, pretreatment with CYD0682 blocked transforming growth factor-β–induced type I collagen and fibronectin production. Conclusions In comparison with oridonin, its novel derivative CYD0682 may act as a more potent antihepatic fibrosis agent.

AB - Background Activated hepatic stellate cells (HSCs) are responsible for excess extracellular matrix (ECM) protein deposition in liver fibrosis. Previously, our group reported that the natural compound oridonin induces apoptosis, inhibits cell proliferation, and downregulates ECM proteins in activated HSC. In this study, the antifibrogenic effects of oridonin derivative CYD0682 on the activated human LX-2 and rat HSC-T6 stellate cell lines were investigated. Methods Cell proliferation was measured by alamarBlue assay. Apoptosis was detected by Cell Death ELISA and staining of Yo-Pro-1 and propidium iodide. Cell cycle was determined by flow cytometry. Immunoblot and immunofluorescence staining were performed for cellular protein expression. Results CYD0682 treatment significantly inhibited LX-2 cell proliferation in a dose- and time-dependent manner with an IC50 value of 0.49 μM for 48 h, ∼10-fold greater potency than oridonin. Similar results were observed in HSC-T6 cells. In contrast, 2.5 μM of CYD0682 treatment had no significant effects on proliferation of the human hepatocyte cell line C3A. CYD0682 treatment induced LX-2 cell apoptosis and S-phase cell cycle arrest and was associated with activation of p53, p21, and cleaved caspase-3. The myofibroblast marker protein α-smooth muscle actin and major ECM proteins type I collagen and fibronectin were markedly suppressed in a time- and dose-dependent fashion by CYD0682. Furthermore, pretreatment with CYD0682 blocked transforming growth factor-β–induced type I collagen and fibronectin production. Conclusions In comparison with oridonin, its novel derivative CYD0682 may act as a more potent antihepatic fibrosis agent.

KW - Derivatives

KW - Fibrosis

KW - Liver

KW - Oridonin

KW - Stellate cells

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