Connective tissue growth factor in hepatocytes is elevated by carbon tetrachloride via STAT3 activation

Chen,Wenhsu; Li,Yingxiao; Hsu,Chao‑Tien; Niu,Chiang‑Shan; Pen,Wen‑Huang; Cheng,Kai‑Chun; Niu,Ho‑Shan

doi:10.3892/mmr.2020.10916

Connective tissue growth factor in hepatocytes is elevated by carbon tetrachloride via STAT3 activation

Authors:
- Wenhsu Chen
- Yingxiao Li
- Chao‑Tien Hsu
- Chiang‑Shan Niu
- Wen‑Huang Pen
- Kai‑Chun Cheng
- Ho‑Shan Niu
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Affiliations: Department of Internal Medicine, E‑Da Hospital, I‑Shou University, Kaohsiung 82401, Taiwan, R.O.C., Department of Psychosomatic Internal Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima 890‑8520, Japan, Department of Pathology, E‑Da Hospital, I‑Shou University, Kaohsiung 82401, Taiwan, R.O.C., Department of Nursing, Tzu Chi University of Science and Technology, Hualien 97005, Taiwan, R.O.C., Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, College of Chinese Medicine, China Medical University, Taichung 40402, Taiwan, R.O.C., Pharmacological Department of Herbal Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima 890‑8520, Japan
Published online on: January 8, 2020 https://doi.org/10.3892/mmr.2020.10916
Pages: 1390-1398

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Abstract

Carbon tetrachloride (CCl4) is widely used to induce hepatic fibrosis. Therapeutic agents alleviate hepatic fibrosis by inhibiting signal transducer and activator of transcription 3 (STAT3) activation. To understand the direct effects of CCl4 on STAT3 expression in the liver, the present study incubated cultured hepatocytes expressing connective tissue growth factor (CTGF) with CCl4. Rats exposed to CCl4 for 8 weeks exhibited hepatic fibrosis, which was confirmed through the assessment of plasma biomarkers. Isolated liver samples were used to determine the protein levels of CTGF and STAT3 using western blotting. In addition, STAT3 expression was silenced in α mouse liver 12 (AML‑12) cells using small interfering RNA transfection. In addition, a pharmacological inhibitor, stattic, was used to inhibit STAT3 expression. The incubation of AML‑12 cells with CCl4 induced a dose‑dependent increase in CTGF expression and STAT3 activation. Notably, silymarin, an extract from milk thistle, inhibited these changes in AML‑12 cells and the antioxidant tiron produced similar effects. Silencing of STAT3 reduced the CTGF expression promoted by CCl4 in the hepatocytes. Additionally, similar to tiron, stattic inhibited CTGF expression induced by CCl4. In conclusion, CCl4 may activate STAT3 through oxidative stress to promote CTGF expression, which is one of the main factors contributing to the risk of hepatic fibrosis.

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