The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, China.
Department of Oral Radiology, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
J Oral Pathol Med. 2018 Oct;47(9):847-855. doi: 10.1111/jop.12765. Epub 2018 Jul 27.
The human salivary gland (HSG) cell line has so far been used as in vitro models for study of the influence of cytokines and pharmacologic agents on salivary glands, as well as a model system for inflammation in Sjögren's syndrome (SS). This study aimed to determine the effect of IL-17 on IL-6 production and the underlying molecular mechanism regulated by the HSG cell line.
Immunofluorescence analyses, RT-PCR, and Western blot were conducted to evaluate the IL-17 receptor (IL-17R) expression in cultured HSG cells. Real-time PCR and ELISA were then utilized to establish the mRNA and protein levels of IL-6 in IL-17-stimulated HSG cells. Western blot, flow cytometry, immunofluorescence, and inhibitor analyses were conducted to elucidate the involved signaling pathways.
The HSG cells reliably expressed the IL-17R mRNA and its encoded surface-bound protein. The expression of IL-6 mRNA and protein was upregulated by stimulation of HSG cells with IL-17; this effect was impeded by IL-17- or IL-17R-neutralizing antibodies. IL-17 stimulation ended up with the fast phosphorylation of p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK), Akt, and translocation of nuclear factor-kappaB (NF-κB) in the HSG cells. p38 MAPK, Akt, and NF-κB inhibitors significantly subdued IL-6 generation in HSG cells stimulated by IL-17. PD98059, an ERK inhibitor, decreased IL-6 generation under low dose of IL-17 but not with high dose.
The HSG cells expressed IL-17R and reacted to IL-17 to generate IL-6 via the stimulation of ERK, p38 MAPK, Akt, and NF-κB signaling pathways.
人唾液腺（HSG）细胞系已被广泛应用于研究细胞因子和药物对唾液腺的影响，以及作为干燥综合征（SS）炎症模型的体外模型。本研究旨在确定 IL-17 对 HSG 细胞系中 IL-6 产生的影响及其潜在的分子调控机制。
通过免疫荧光分析、RT-PCR 和 Western blot 评估培养的 HSG 细胞中 IL-17 受体（IL-17R）的表达。然后利用实时 PCR 和 ELISA 检测 IL-17 刺激 HSG 细胞后 IL-6 的 mRNA 和蛋白水平。通过 Western blot、流式细胞术、免疫荧光和抑制剂分析来阐明涉及的信号通路。
HSG 细胞可靠地表达了 IL-17R mRNA 及其编码的表面结合蛋白。IL-17 刺激 HSG 细胞可上调 IL-6 mRNA 和蛋白的表达；这种作用可被 IL-17 或 IL-17R 中和抗体所抑制。IL-17 刺激导致 HSG 细胞中 p38 丝裂原活化蛋白激酶（MAPK）、细胞外信号调节激酶（ERK）、Akt 和核因子-κB（NF-κB）的快速磷酸化。p38 MAPK、Akt 和 NF-κB 抑制剂可显著抑制 IL-17 刺激的 HSG 细胞中 IL-6 的产生。ERK 抑制剂 PD98059 在低剂量 IL-17 下降低 IL-6 的产生，但在高剂量下则无此作用。
HSG 细胞表达 IL-17R，并通过 ERK、p38 MAPK、Akt 和 NF-κB 信号通路对 IL-17 刺激产生 IL-6。
