Interleukin‑10 promotes primary rat hepatic stellate cell senescence by upregulating the expression levels of p53 and p21

Huang,Yue‑Hong; Chen,Ming‑Hua; Guo,Qi‑Lan; Chen,Yun‑Xin; Zhang,Li‑Juan; Chen,Zhi‑Xin; Wang,Xiao‑Zhong

doi:10.3892/mmr.2018.8592

Interleukin‑10 promotes primary rat hepatic stellate cell senescence by upregulating the expression levels of p53 and p21

Authors:
- Yue‑Hong Huang
- Ming‑Hua Chen
- Qi‑Lan Guo
- Yun‑Xin Chen
- Li‑Juan Zhang
- Zhi‑Xin Chen
- Xiao‑Zhong Wang
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Affiliations: Department of Gastroenterology, Fujian Medical University Union Hospital, Fuzhou, Fujian 350001, P.R. China
Published online on: February 12, 2018 https://doi.org/10.3892/mmr.2018.8592
Pages: 5700-5707
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Liver fibrosis is characterized by the excessive deposition of extracellular matrix (ECM) components, and activated hepatic stellate cells (HSCs) are a primary source of ECM. Several studies have revealed that the induction of HSC senescence may reduce liver fibrosis. The effect of interleukin‑10 (IL‑10) on the senescence of activated HSCs is not fully understood. Therefore, the present study examined its effects and potential mechanisms in activated primary rat HSCs. Collagenase perfusion and density gradient centrifugation methods were used to isolate rat HSCs. HSCs were identified by autofluorescence, Oil Red O staining and immunocytochemical analysis. Activated HSCs were treated with 0, 10, 20 or 40 ng/ml IL‑10 for 24 h. Senescence‑associated β‑galactosidase (SA‑β‑Gal) staining, flow cytometry analysis and a cell counting kit‑8 assay were performed to detect the senescence, apoptosis and viability of rat HSCs, respectively. Reverse transcription‑quantitative polymerase chain reaction, western blot analysis and enzyme linked immunosorbent assays were used to detect the expression of senescence‑associated proteins and cytokines. Freshly isolated rat HSCs exhibited a striking blue‑green autofluorescence and HSC retinoid droplets were stained bright red by Oil Red O. Immunocytochemical analysis demonstrated the cytoplasmic expression of HSC markers desmin and α‑smooth muscle actin. The number of SA‑β‑Gal positive HSCs, the apoptotic rate and the expression levels of p53, p21 and tumor necrosis factor‑α were significantly increased following IL‑10 treatment. HSC viability and IL‑6 and IL‑8 expression levels were significantly decreased compared with the control group. In summary, primary rat HSCs were successfully isolated and IL‑10 was demonstrated to promote the senescence of activated primary rat HSCs through the upregulation of p53 and p21 expression.

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