| 庄同田,易秀莉,亢 盼,陈佳希,李舒丽.病毒感染对角质形成细胞CCL20表达的影响及其机制[J].,2020,(10):1840-1845 |
| 病毒感染对角质形成细胞CCL20表达的影响及其机制 |
| Impacts and Mechanism of Virus Infection on Expression of CCL20 in Keratinocytes |
| 投稿时间：2019-10-28 修订日期：2019-11-24 |
| DOI：10.13241/j.cnki.pmb.2020.10.009 |
| 中文关键词: 病毒 Poly(I:C) 角质形成细胞 CCL20 |
| 英文关键词: Virus Poly(I:C) Keratinocyte CCL20 |
| 基金项目:国家自然科学基金项目（81803122） |
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| 中文摘要: |
| 摘要 目的：既往研究发现，趋化因子CCL20在银屑病、白癜风等在内的多种自身免疫性皮肤疾病的病理过程中扮演了重要的角色，同时病毒感染也被认为是自身免疫性疾病的重要参与者。皮肤组织是机体抵御外界病原体的第一道屏障，其中角质形成细胞被认为在启动免疫中发挥了关键的作用。视黄酸诱导基因蛋白I（RIG-I ）是固有免疫模式识别受体家族的重要成员，其能够被病毒复制的中间产物激活。然而，病毒感染是否会通过RIG-I信号通路影响角质形成细胞中CCL20的表达，进而参与自身免疫性皮肤疾病的病理过程仍不清楚。本文使用聚肌胞苷酸(Poly(I:C))来体外模拟病毒感染，探究病毒感染对皮肤角质形成细胞CCL20表达的影响，并且通过小干扰RNA沉默关键分子来探究相应的分子机制。方法：首先，体外细胞实验使用Poly(I:C)刺激角质形成细胞系HaCaT，通过Western-blot实验和qRT-PCR实验探究Poly(I:C)对HaCat中RIG-I表达的影响；接下来，通过实时荧光定量PCR(qRT-PCR)以及酶联免疫吸附测定实验（ELISA）检测Poly(I:C)对角质形成细胞CCL20分泌的影响；线粒体抗病毒信号蛋白(MAVS)在RIG-I的下游发挥着重要作用，我们通过小干扰RNA（si-RNA）阻断RIG-I-MAVS-NF-κB信号通路关键分子，探究Poly(I:C)诱导角质形成细胞CCL20表达升高的分子机制。结果：Poly(I:C)能够明显促进角质形成细胞中RIG-I的表达及CCL20的表达和分泌；Poly(I:C)诱导角质形成细胞CCL20分泌是由RIG-I-MAVS-NF-κB信号通路介导的。结论：Poly(I:C)模拟病毒感染能够通过RIG-I-MAVS-NF-κB信号通路介导CCL20表达增加，进而参与自身免疫性皮肤疾病的病理过程。 |
| 英文摘要: |
| ABSTRACT Objective: Previous studies have validated that CCL20 played a key role in the pathogenesis of many autoimmune skin diseases including psoriasis and vitiligo. Meanwhile, virus infection also is considered as an important participant factor of autoimmune diseases. Skin tissue, as the outermost layer in organism, contributes critically to the pathogens defense and elimination. Particularly, keratinocyte, accounting for the absolutely percentage in epidermis, is regarded as the important executor in the process of initiating immunology. RIG-I （Retinoic acid-inducible gene I, RIG-I）is an innate pathogen recognition receptor (PRR) and could be activated by the virus replication intermediate products. However, whether virus infection could influence the secretion of CCL20 in keratinocytes via RIG-I signaling pathway and forwardly to aggravate pathogenesis of autoimmune skin diseases is still elusive. In the present study, using Poly(I:C) to imitate virus infection in vitro, we explored the impact of virus infection on the expression of CCL20 in keratinocytes. In addition, using the corresponding small interference RNA (si-RNA) to block the key molecule, we explored the underlying mechanism. Methods: Firstly, using Poly(I:C) to stimulate keratinocyte cell line HaCat, we explored the effect of Poly(I:C) on the expression of RIG-I by Western-blot assay and qRT-PCR assay. Sequentially, using qRT-PCR and ELISA to validate the impacts of Poly(I:C) on the expression of CCL20 in HaCaT cells in vitro. In addition, MAVS（Mitochondria anti-virus signaling protein, MAVS）functions in the downstream of RIG-I, using small interference RNA (si-RNA) to impede key signaling components in RIG-I-MAVS-NF-κB signaling pathway to determine the specific mechanism of how Poly(I:C) induces the expression of CCL20. Results: Poly(I:C) could induce the expression of RIG-I and the secretion of CCL20 in keratinocytes. The RIG-I-MAVS-NF-κB signaling pathway contributes significantly to the secretion of CCL20 in response to Poly(I:C). Conclusion: Virus invasion to skin could induce the secretion of CCL20 via RIG- I-MAVS-NF-κB signaling pathway in keratinocytes and forwardly to participate in the pathogenesis of autoimmune skin diseases. |
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