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

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.

Cellular Interplay as a Consequence of Inflammatory Signals Leading to Liver Fibrosis Development

“…IL-10 is involved in downregulating the pro-inflammatory processes in liver fibrosis [7]. In vivo studies showed that IL-10 inhibits the expression of aHSCs markers demonstrating its anti-fibrotic effects [97,98]. In a more recent study, IL-10 was shown to promote cellular death of aHSCs by senescence and upregulation of p53 and p21 expression [98].…”

“…In vivo studies showed that IL-10 inhibits the expression of aHSCs markers demonstrating its anti-fibrotic effects [97,98]. In a more recent study, IL-10 was shown to promote cellular death of aHSCs by senescence and upregulation of p53 and p21 expression [98]. Another anti-inflammatory cytokine is IL-22 that belongs to the IL-10 family and it is produced by innate immune system cells [99][100][101].…”

Silencing p53 inhibits interleukin 10-induced activated hepatic stellate cell senescence and fibrotic degradation in vivo

“…Recently, we found that IL-10 promoted primary rat HSCs senescence by up-regulating the expression of p53 and p21 in vitro. 20 Moreover, we demonstrated that induction of HSCs senescence by IL-10 may be associated with p53-mediated signaling pathway in vitro . 21 As p53 plays a vital role in the senescence of activated HSCs mediated by IL-10 and then affect the progression of fibrosis, we sought to determine whether p53 knockdown could affect the senescence of activated HSCs and degradation of fibrosis mediated by IL-10.…”

Pro- and Anti-fibrogenic Functions of Gram-Negative Bacterial Lipopolysaccharide in the Liver

“…Elimination of the injury stimulus causes aHSCs to undergo apoptosis (72), senescence (73), or reversal to quiescent or the so-called "inhibited phenotype" (iHSC) leading to regression of fibrosis (8,13,(74)(75)(76). IL10 and IL22 can be critically involved in the fibrosis reversal process as evidenced by IL10-induced inhibition of the expression of the activation markers in aHSCs (77)(78)(79), and IL10-and IL22-induced aHSC death by senescence (80,81). It is important to note that iHSCs can be rapidly reactivated upon return of the injury stimulus causing accelerated development of fibrosis (75).…”