Effects of fluoxetine on TLR2/NF-κB signaling pathway and inflammatory factors in human conjunctival epithelial cells

doi:10.12092/j.issn.1009-2501.2019.11.003

Abstract

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

CLC Number:

R965.2
R777

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.

References

［1］Ling L, Qian G, Flanagan J, et al. Genetic factors and molecular mechanisms in dry eye disease［J］. Ocul Surf, 2018, 16(2): 206-217.
［2］Kocer E, Kocer A, Ozsutcu M, et al. Dry eye related to commonly used new antidepressants［J］. J Clin Psychopharmacol, 2015, 35(4): 411-413.
［3］Wen W, Wu Y, Chen Y, et al. Dry eye disease in patients with depressive and anxiety disorders in Shanghai［J］. Cornea, 2012, 31(6): 686-692.
［4］Paduch R, Matysikwozniak A, Maciejewski R, et al. Paracrine interactions between the conjunctival and corneal epithelial cells regulate microenvironmental homeostasis during artificially induced inflammation［J］. Curr Eye Res, 2018, 43(5): 611-620.
［5］Yang Y, Huang C, Lin X, et al. 0.005% Preservative-free latanoprost induces dry eye-like ocular surface damage via promotion of inflammation in mice［J］. Invest Ophthalmol Vis Sci, 2018, 59(8): 3375-3384.
［6］Brignole F, Pisella PJ, Goldschild M, et al. Flow cytometric analysis of inflammatory markers in conjunctival epithelial cells of patients with dry eyes［J］. Invest Ophthalmol Vis Sci, 2014, 41(6): 1356-1363.
［7］Ma SQ, Wei HL, Zhang X. TLR2 regulates allergic airway inflammation through NF-kappaB and MAPK signaling pathways in asthmatic mice［J］. Eur Rev Med Pharmacol Sci, 2018, 22(10): 3138-3146.
［8］Yu Y,Chen Y,Wang Y,et al.TLR2/MyD88/NF-kappaB signaling pathway regulates IL-1beta production in DF-1cells exposed to mycoplasma gallisepticum LAMPs［J］.Microb Pathog,2018,117(1):225-231.
［9］Chang H, Lai P, Weng J, et al. Toll-like receptor 2-mediated NF-κB inflammatory responses in dry eye associated with cGVHD［J］. Mol Vis, 2011, 17(1): 2605-2611.
［10］Bertolin M, Breda C, Ferrari S, et al. Optimized protocol for regeneration of the conjunctival epithelium using the cell suspension technique［J］. Cornea, 2019, 38(4): 469-479.
［11］Hunt D, Drake LA, Drake JR. Murine macrophage TLR2-FcγR synergy via FcγR licensing of IL-6 cytokine mRNA ribosome binding and translation［J］. Plos One, 2018, 13(7): e0200764.
［12］Gemmel M, Bogi E, Ragan C, et al. Perinatal selective serotonin reuptake inhibitor medication (SSRI) effects on social behaviors, neurodevelopment and the epigenome［J］. Neurosci Biobehav Rev, 2018, 85(1): 102-116.
［13］Mandrioli R,Protti M,Mercolini L.New-generation,non-ssri antidepressants:therapeutic drug monitoring and pharmacological interactions.Part 1:SNRIs,SMSs,SARIs［J］.Curr Med Chem,2018,25(7):772-792.
［14］Mrugacz M, Ostrowska L, Bryl A, et al. Pro-inflammatory cytokines associated with clinical severity of dry eye disease of patients with depression［J］. Adv Med Sci, 2017, 62(2): 338-344.
［15］Rhee MK, Mah FS. Inflammation in dry eye disease: how do we break the cycle ［J］? Ophthalmology, 2017, 124(11s): 14-19.
［16］Ma S, Yu Z, Feng S, et al. Corneal autophagy and ocular surface inflammation: A new perspective in dry eye［J］. Exp Eye Res, 2019, 184(1): 126-134.
［17］Zhang C, Xi L, Zhao S, et al. Interleukin-1beta and tumour necrosis factor-alpha levels in conjunctiva of diabetic patients with symptomatic moderate dry eye: case-control study［J］. BMJ Open, 2016, 6(8): e010979.
［18］Lee SY, Han SJ, Nam SM, et al. Analysis of tear cytokines and clinical correlations in Sjogren syndrome dry eye patients and non-Sjogren syndrome dry eye patients［J］. Am J Ophthalmol, 2013, 156(2): 247-253, e241.
［19］Li K, Zhang C, Yang Z, et al. Evaluation of a novel dry eye model induced by oral administration of finasteride［J］. Mol Med Rep, 2017, 16(6): 8763-8770.
［20］Du J, Chi Y, Song Z, et al. Crocin reduces Aspergillus fumigatus-induced airway inflammation and NF-kappaB signal activation［J］. J Cell Biochem, 2018, 119(2): 1746-1754.
［21］Park JY, Chung TW, Jeong YJ, et al. Ascofuranone inhibits lipopolysaccharide-induced inflammatory response via NF-kappaB and AP-1, p-ERK, TNF-α, IL-6 and IL-1β in RAW 264.7 macrophages［J］. Plos One, 2017, 12(2): e0171322.
［22］Chen KY, Wang LC. Stimulation of IL-1β and IL-6 through NF-κB and sonic hedgehog-dependent pathways in mouse astrocytes by excretory/secretory products of fifth-stage larval Angiostrongylus cantonensis［J］. Parasit Vectors, 2017, 10(1): 445-452.
［23］Gao H,Liu L,Zhao Y,et al.Human IL-6,IL-17,IL-1β,and TNF-α differently regulate the expression of pro-inflammatory related genes,tissue factor,and swine leukocyte antigen class I in porcine aortic endothelial cells［J］.Xenotransplantation,2017,24(2):e12291.
［24］Yang JM, Rui BB, Chen C, et al. Acetylsalicylic acid enhances the anti-inflammatory effect of fluoxetine through inhibition of NF-kappaB, p38-MAPK and ERK1/2 activation in lipopolysaccharide-induced BV-2 microglia cells［J］. Neuroscience, 2014, 275(1): 296-304.
［25］Song T, Li H, Tian Z, et al. Disruption of NF-kappaB signaling by fluoxetine attenuates MGMT expression in glioma cells［J］. Onco Targets Ther, 2015, 8(1): 2199-2208.

[1]	DU Caifeng, MAO Yunan, GAO Jia, WANG Juan, LI Xiaoye, TIAN Junsheng, LIU Hongqi. Research on serum metabolic markers of menopausal syndrome based on 1H-NMR metabolomics [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 961-968.
[2]	LIAO Mengling, WANG Yan, LUO Jing, WANG Nuoyan, HUA Ling, ZHANG Yu, DENG Fei, YUAN Yue, ZHOU Jun, ZHOU Hong. Molecular mechanism of artesunate attenuates the release of proinflammatory cytokines from macrophages [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 969-978.
[3]	LIU Lijing, QIAN Hong, MENG Qingxin, ZHANG Xiang, HE Bin, HE Jianbin, WEI Yingmin. Sinomenine inhibits oxidative stress and pulmonary fibrosis by activating the Keap1/Nrf2 signaling pathway [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 979-987.
[4]	YOU Rongli, HUANG Yurong, MAORui, HAI Lina, WANG Yingli, WANG Yan. Mechanism of compound kushen injection in the treatment of lung cancer based on serum metabolomics and network pharmacology [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 988-999.
[5]	JIANG Daoli, CHOU Xiaohua, LIU Zhidong, LI Wei, ZHANG Bo, XU Sang, TAN Defei, FANG Yi, LV Dongmei, WANG Tao. Real-world study of ceftazidime-avibactam in the treatment of multidrug-resistant gram-negative bacterial infections [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 1008-1017.
[6]	MA Yan, TIAN Gaopeng, SHI Xingwen, SUN Ting, XIE Jingjing, ZHEN Donghu. Correlation between 25(OH)D and metabolically related fatty liver in T2DM population [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 1018-1026.
[7]	WU Yujie, ZHAO Chengcheng, XI Qing. Evaluation of tegacycline regimens in treatment of gram-negative bacterial infections with Monte Carlo simulation [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 1027-1033.
[8]	XU Yiqi, WU Qian, LIU Shu, LIU Fan, XING Chunyan, LI Qin, HE Junjun, HE Chunling, ZHAO Yongli, GAO Jialin. Clinical efficacy and anti-inflammation/anti-fibrosis effect of tripterygium glycosides in the treatment of diabetic nephropathy [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 1034-1042.
[9]	XIAO Yue, LI Tangfei, XUE Qianfu. Analysis of the efficacy and safety of Peg-IFN-α in the treatment of essential thrombocythemia [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 1049-1055.
[10]	MAO Jun, RONG Yu, LI Yangen, XIONG Dengxi, ZHAO Peng, ZHA Zhengjiang. Effect and safety of flurbiprofen axetil in the analgesic treatment of patients with craniocerebral injury [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 1056-1060.
[11]	HUA Zhihui, LIU Dong, ZHANG Jiandong, CAI Siyu, GE Chunli, CHEN Nan, QI Qi. Clinical evaluation of the efficacy against helicobacter pylori by amoxicillin from volume-based procurement and potassium amoxicillin clavulanate [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 1061-1066.
[12]	WANG Jie, LI Long, CHEN Feng, LIU Shengfei. Research progress of nuclear factor erythroid 2-related factor 2 in pulmonary fibrosis [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(9): 1067-1072.
[13]	HE Lihua, ZHU Xiuzhi, JIANG Yizhou. Research progress on immunotherapy for triple-negative breast cancer [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(8): 842-853.
[14]	ZHANG Huyunlong, ZHU Xiuzhi, JIN Xi, SHAO Zhimin. Mechanisms of endocrine-resistance and therapeutic breakthroughs in hormone receptor-positive, HER2-negative breast cancer [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(8): 854-865.
[15]	FU Xiaoyu, KONG Deguang, LI Juanjuan. Treatment progress of triple positive breast cancer [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(8): 866-875.