State Key Laboratory of Analytical Chemistry for Life Science, School of Life Sciences, Nanjing University, Nanjing, China.
State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, China.
Inflamm Bowel Dis. 2021 Oct 20;27(11):1821-1831. doi: 10.1093/ibd/izab072.
Interleukin-10 (IL-10) is a potent immunoregulatory cytokine that plays a pivotal role in maintaining mucosal immune homeostasis. As a novel synthetic inhibitor of salt-inducible kinases (SIKs), HG-9-91-01 can effectively enhance IL-10 secretion at the cellular level, but its in vivo immunoregulatory effects remain unclear. In this study, we investigated the effects and underlying mechanism of HG-9-91-01 in murine colitis models.
The anti-inflammatory effects of HG-9-91-01 were evaluated on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-, dextran sulfate sodium-induced colitis mice, and IL-10 knockout chronic colitis mice. The in vivo effector cell of HG-9-91-01 was identified by fluorescence-activated cell sorting and quantitative real-time polymerase chain reaction. The underlying mechanism of HG-9-91-01 was investigated via overexpressing SIKs in ANA-1 macrophages and TNBS colitis mice.
Treatment with HG-9-91-01 showed favorable anticolitis effects in both TNBS- and DSS-treated mice through significantly promoting IL-10 expression in colonic macrophages but failed to protect against IL-10 KO murine colitis. Further study indicated that HG-9-91-01 markedly enhanced the nuclear level of cAMP response element-binding protein (CREB)-regulated transcription coactivator 3 (CRTC3), whereas treatment with lentiviruses encoding SIK protein markedly decreased the nuclear CRTC3 level in HG-9-91-01-treated ANA-1 macrophages. In addition, intracolonic administration with lentiviruses encoding SIK protein significantly decreased the nuclear CRTC3 level in the lamina propria mononuclear cells and ended the anti-inflammatory activities of HG-9-91-01.
We found that HG-9-91-01 promoted the IL-10 expression of colonic macrophages and exhibited its anticolitis activity through the SIK/CRTC3 axis, and thus it may represent a promising strategy for inflammatory bowel disease therapy.
白细胞介素-10（IL-10）是一种有效的免疫调节细胞因子，在维持黏膜免疫稳态中发挥关键作用。作为新型盐诱导激酶（SIKs）的合成抑制剂，HG-9-91-01 可以在细胞水平上有效增强 IL-10 的分泌，但它在体内的免疫调节作用尚不清楚。在这项研究中，我们研究了 HG-9-91-01 在小鼠结肠炎模型中的作用及其潜在机制。
用 2,4,6-三硝基苯磺酸（TNBS）、葡聚糖硫酸钠诱导的结肠炎小鼠和 IL-10 基因敲除慢性结肠炎小鼠评估 HG-9-91-01 的抗炎作用。通过荧光激活细胞分选和实时定量聚合酶链反应鉴定 HG-9-91-01 的体内效应细胞。通过在 ANA-1 巨噬细胞和 TNBS 结肠炎小鼠中转染 SIKs 来研究 HG-9-91-01 的潜在机制。
HG-9-91-01 治疗在 TNBS 和 DSS 处理的小鼠中均显示出良好的抗结肠炎作用，通过显著促进结肠巨噬细胞中 IL-10 的表达，但未能预防 IL-10 KO 小鼠的结肠炎。进一步的研究表明，HG-9-91-01 显著增强了环磷酸腺苷反应元件结合蛋白（CREB）调节转录共激活因子 3（CRTC3）的核水平，而用编码 SIK 蛋白的慢病毒处理则显著降低了 HG-9-91-01 处理的 ANA-1 巨噬细胞中的核 CRTC3 水平。此外，经腔内给予编码 SIK 蛋白的慢病毒可显著降低固有层单核细胞中的核 CRTC3 水平，并终止 HG-9-91-01 的抗炎作用。
我们发现 HG-9-91-01 通过 SIK/CRTC3 轴促进结肠巨噬细胞中 IL-10 的表达，并表现出其抗结肠炎活性，因此它可能代表一种有前途的炎症性肠病治疗策略。
