Open and Key laboratory of Hepatobiliary & Pancreatic Surgery and Digestive Organ Transplantation at Henan Universities, Zhengzhou, Henan, People's Republic of China.
ZhengZhou Key Laboratory of Hepatobiliary & Pancreatic Diseases and Organ Transplantation, Zhengzhou, Henan, People's Republic of China.
Mol Biol Rep. 2021 Feb;48(2):1687-1695. doi: 10.1007/s11033-020-06090-2. Epub 2021 Jan 23.
Ischemia-reperfusion injury (IRI) is inevitable during liver surgery, and it is an important factor affecting the prognosis of patients. IL-6 rs1800796 single nucleotide polymorphisms (SNPs) can promote synthesis and secretion of IL-6 and protect hepatocytes from IRI. In this study, we investigated the mechanisms by which IL-6 alleviates hepatic IRI. We transfected lentivirus which carries IL-6 rs1800796 to L02 cells and constructed the cell line (L02-IL6) with a high expression of IL-6. The biological function of IL-6 SNPs was explored through a cell model of hypoxia-reoxygenation (H/R). Cell viability was evaluated by CCK8 and Real-Time Cell Analysis (RTCA), and found that the viability of the L02-IL6 cells was higher than that of the control group (P < 0.01). Flow cytometry assay showed that the rate of apoptosis was significantly decreased in L02-IL6 cells. Furthermore, in comparison with the control group, the level of cleaved-caspase3, which is an important marker of apoptosis, was dramatically decreased. These differences showed that the sequence variants at rs1800796 of the IL-6 gene could improve the resistance against H/R. Moreover, the levels of autophagy-related proteins, such as LC3 and Beclin-1, were upregulated in L02-IL6 group on H/R injury, which means IL-6 could alleviate apoptosis via activating the autophagy pathway. And we also found that the STAT3 signal pathway was activated. Next, we investigated whether the exogenous treatment with IL-6 affect hepatocytes and thus play a protective role. We pre-treated the L02 cells with recombinant human IL-6 for 12 h and then made H/R treatment. We found the treatment with 100 ng/ml IL-6 alleviated the damage of L02 cells and inhibited the apoptosis. And the further study revealed the pre-treatment with IL-6 activated the STAT3 signaling pathway in the L02 cells and then caused the activation of autophagy and apoptosis inhibition. IL-6 might play a critical role in alleviating hepatic IRI, through its modulation of the STAT3 signaling pathway, and activation of autophagy. Recombinant human IL-6 might be a potential therapeutic target in hepatic IRI.
缺血再灌注损伤（IRI）是肝外科手术中不可避免的，是影响患者预后的重要因素。IL-6 rs1800796 单核苷酸多态性（SNP）可促进 IL-6 的合成和分泌，保护肝细胞免受 IRI。在本研究中，我们研究了 IL-6 减轻肝 IRI 的机制。我们转染携带 IL-6 rs1800796 的慢病毒，构建了 IL-6 高表达的细胞系（L02-IL6）。通过缺氧复氧（H/R）细胞模型探讨了 IL-6 SNPs 的生物学功能。通过 CCK8 和实时细胞分析（RTCA）评估细胞活力，发现 L02-IL6 细胞的活力高于对照组（P<0.01）。流式细胞术检测显示，L02-IL6 细胞凋亡率明显降低。此外，与对照组相比，凋亡的重要标志物 cleaved-caspase3 的水平显著降低。这些差异表明，IL-6 基因 rs1800796 序列变异可提高对 H/R 的抵抗力。此外，在 H/R 损伤时，L02-IL6 组自噬相关蛋白 LC3 和 Beclin-1 的水平上调，表明 IL-6 可通过激活自噬途径减轻凋亡。并且我们还发现 STAT3 信号通路被激活。接下来，我们研究了外源性 IL-6 处理是否会影响肝细胞并发挥保护作用。我们用重组人 IL-6 预处理 L02 细胞 12 h，然后进行 H/R 处理。我们发现，用 100 ng/ml IL-6 处理可减轻 L02 细胞的损伤并抑制凋亡。进一步的研究表明，IL-6 预处理激活了 L02 细胞中的 STAT3 信号通路，进而激活自噬并抑制凋亡。IL-6 可能通过调节 STAT3 信号通路和激活自噬在减轻肝 IRI 中发挥关键作用。重组人 IL-6 可能是肝 IRI 的潜在治疗靶点。
