氟西汀对人结膜上皮细胞TLR2/NF-κB信号通路和炎性因子的影响

doi:10.12092/j.issn.1009-2501.2019.11.003

摘要/Abstract

摘要：

目的:观察氟西汀对人结膜上皮细胞Toll样受体2（toll-like receptor 2，TLR2）/核转录因子-κB（NF-κB）信号通路和炎性因子的影响，探讨五羟色胺再摄取抑制剂（selective serotonin reuptake inhibitor，SSRI）引起干眼症的潜在机制。方法:将结膜上皮细胞分为空白对照组和药物组，药物组采用氟西汀进行干预。采用CCK-8法明确氟西汀对人结膜上皮细胞半数抑制率作为后续实验浓度。采用RT-PCR法检测TLR2、NF-κB、白细胞介素（interleukin，IL）-1β、IL-2、IL-6和肿瘤坏死因子（tumor necrosis factor，TNF）-α的mRNA水平；免疫荧光法和Western blot法检测TLR2和NF-κB的蛋白表达水平；ELISA法检测IL-1β、IL-2、IL-6和TNF-α的蛋白表达水平。结果:10-9～10-5mol/L范围内氟西汀对结膜上皮细胞具有增殖抑制作用，且具有浓度依赖性。RT-PCR法检测结果显示，氟西汀组TLR2、NF-κB、IL-1β、IL-2、IL-6和TNF-α mRNA水平均高于空白对照组（P＜0.05）。药物组TLR2和NF-κB的累积光密度（IOD）值明显高于空白对照组（P＜0.05），提示药物组TLR2和NF-κB蛋白表达量高于空白对照组。Western blot法检测结果显示药物组TLR2和NF-κB的蛋白表达水平均高于空白对照组（P＜0.05）。ELISA检测结果显示药物组IL-1β、IL-2、IL-6和TNF-α蛋白表达水平均高于空白对照组（P＜0.05）。结论:本研究发现氟西汀可能通过激活结膜上皮细胞TLR2/NF-κB信号通路，促进炎症因子水平，这可能是氟西汀所致干眼症潜在的生物学机制。

关键词: 干眼症, 氟西汀, 人结膜上皮细胞, Toll样受体2, 核转录因子-κB, 炎症

Abstract:

AIM: To observe the effects of fluoxetine on Toll-like receptor 2 (TLR2)/nuclear transcription factor-kappa B (NF-κB) signaling pathway and inflammatory factors in human conjunctival epithelial cells, and to explore the potential mechanism of SSRI-induced xerophthalmia. METHODS: Conjunctival epithelial cells were divided into blank control group and drug group. Fluoxetine was used to intervene in drug group. CCK-8 method was used to determine the half inhibition rate of fluoxetine on human conjunctival epithelial cells as a follow-up concentration. The levels of TLR2, NF-kappa B, interleukin (IL) -1beta, IL-2, IL-6 and tumor necrosis factor (TNF) -alpha were detected by RT-PCR, the levels of TLR2 and NF-kappa B protein were detected by immunofluorescence and Western-blot, and the levels of IL-1beta, IL-2, IL-6 and TNF-alpha protein were detected by ELISA. RESULTS:Fluoxetine can inhibit the proliferation of conjunctival epithelial cells in a concentration-dependent manner in the range of 10-9 to 10-5 mol/L. RT-PCR results showed that the levels of TLR2, NF-kappa B, IL-1beta, IL-2, IL-6 and TNF-alpha in fluoxetine group were higher than those in blank control group (P＜0.05). The IOD values of TLR2 and NF-kappa B in the drug group were significantly higher than those in the blank control group (P＜0.05), suggesting that the expressions of TLR2 and NF-kappa B in the drug group were higher than those in the blank control group. Western blot analysis showed that the expression levels of TLR2 and NF-kappa B in the drug group were higher than those in the blank control group (P＜0.05). ELISA results showed that the expression levels of IL-1beta, IL-2, IL-6 and TNF-alpha in the drug group were higher than those in the blank control group (P＜0.05). CONCLUSION: Fluoxetine may promote the level of inflammatory factors by activating TLR2/NF- κB signaling pathway in conjunctival epithelial cells. This may be the potential biological mechanism of fluoxetine-induced xerophthalmia.

Key words: xerophthalmia, fluoxetine, human conjunctival epithelial cells, Toll-like receptor 2, nuclear factor-&kappa, B, inflammation

中图分类号:

R965.2
R777

杨青霞，毛怡清，张燕芳，薄旭芳. 氟西汀对人结膜上皮细胞TLR2/NF-κB信号通路和炎性因子的影响[J]. 中国临床药理学与治疗学, 2019, 24(11): 1227-1233.

YANG Qingxia，MAO Yiqing，ZHANG Yanfang，BO Xufang. Effects of fluoxetine on TLR2/NF-κB signaling pathway and inflammatory factors in human conjunctival epithelial cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(11): 1227-1233.

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