Interleukin‑10 promotes primary rat hepatic stellate cell senescence by upregulating the expression levels of p53 and p21.
Interleukin‑10 promotes primary rat hepatic stellate cell senescence by upregulating the expression levels of p53 and p21.
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Interleukinα10 通过上调 p53 和 p21 的表达水平促进原代大鼠肝星状细胞衰老。
DOI：
10.3892/mmr.2018.8592
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发表时间：
2018-04
影响因子：
3.4
通讯作者：
Wang XZ
中科院分区：
文献类型：
--
作者：
Huang YH;Chen MH;Guo QL;Chen YX;Zhang LJ;Chen ZX;Wang XZ
肝纤维化的特征是细胞外基质（ECM）成分的过度沉积，而活化的肝星状细胞（HSC）是ECM的主要来源。一些研究表明，诱导HSC衰老可以减轻肝纤维化。白细胞介素-10（IL-10）对活化的HSC衰老的影响尚未完全了解。因此，本研究探讨了其在活化的原代大鼠HSC中的作用和潜在机制。胶原酶灌注法和密度梯度离心法分离大鼠HSC。采用自体荧光、油红O染色和免疫细胞化学方法鉴定HSC。用0、10、20或40 ng/ml IL-10处理活化的HSC 24 h。采用衰老相关β-半乳糖苷酶（SA-β-Gal）染色、流式细胞仪分析和细胞计数kit-8法分别检测大鼠HSC的衰老、凋亡和存活率。采用逆转录-定量聚合酶链反应（RT-PCR）、免疫印迹（Western blot）和酶联免疫吸附试验（ELISA）检测衰老相关蛋白和细胞因子的表达。新鲜分离的大鼠肝星状细胞显示出明显的蓝绿色自发荧光，视黄酸滴被油红O染成鲜红色。免疫细胞化学分析显示HSC标志物结蛋白和α-平滑肌肌动蛋白在细胞质中表达。IL-10处理后，SA-β-Gal阳性的HSC数量、凋亡率及p53、p21和TNF-α的表达水平均明显增加。与对照组相比，HSC活力和IL-6和IL-8表达水平显著降低。总之，成功地分离了原代大鼠HSC，并证明IL-10通过上调p53和p21表达促进活化的原代大鼠HSC的衰老。
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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影响因子：
4.6
作者：
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DOI：
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发表时间：
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影响因子：
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影响因子：
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作者：
Jin, Huanhuan;Lian, Naqi;Zheng, Shizhong
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