氟西汀调节TLR4/NF-κB/NLRP3炎症体信号通路改善CUMS大鼠抑郁样行为

doi:10.11958/20220674

天津医药 ›› 2023, Vol. 51 ›› Issue (2): 160-165.doi: 10.11958/20220674

氟西汀调节TLR4/NF-κB/NLRP3炎症体信号通路改善CUMS大鼠抑郁样行为

吕霞(), 黄丽, 张美琳, 樊珺婷, 马泽微, 刘欢^△()

天津医科大学公共卫生学院营养与食品卫生学系，天津市环境营养与人群健康重点实验室（邮编300070）

收稿日期:2022-05-30 修回日期:2022-08-20 出版日期:2023-02-15 发布日期:2023-02-24
通讯作者: ^△E-mail：liuhuan@tmu.edu.cn
作者简介:吕霞（1995），女，硕士在读，主要从事营养与神经科学方面研究。E-mail：2655646457@qq.com
基金资助:
国家自然科学基金资助项目(82173516)

Fluoxetine improves depression-like behavior in CUMS rats by regulating TLR4/NF-κB/NLRP3 inflammasome signaling pathway

LYU Xia(), HUANG Li, ZHANG Meilin, FAN Junting, MA Zewei, LIU Huan^△()

Department of Nutrition and Food Science, School of Public Health, Tianjin Medical University; Tianjin Key Laboratory of Environment, Nutrition, and Public Health, Center for International Collaborative Research on Environment, Nutrition and Public Health, Tianjin 300070, China

Received:2022-05-30 Revised:2022-08-20 Published:2023-02-15 Online:2023-02-24
Contact: ^△E-mail：liuhuan@tmu.edu.cn

吕霞, 黄丽, 张美琳, 樊珺婷, 马泽微, 刘欢. 氟西汀调节TLR4/NF-κB/NLRP3炎症体信号通路改善CUMS大鼠抑郁样行为[J]. 天津医药, 2023, 51(2): 160-165.

LYU Xia, HUANG Li, ZHANG Meilin, FAN Junting, MA Zewei, LIU Huan. Fluoxetine improves depression-like behavior in CUMS rats by regulating TLR4/NF-κB/NLRP3 inflammasome signaling pathway[J]. Tianjin Medical Journal, 2023, 51(2): 160-165.

摘要/Abstract

摘要：

目的基于Toll样受体4（TLR4）/核因子κB（NF-κB）/NOD样受体热蛋白结构域相关蛋白3（NLRP3）炎症体信号通路探究氟西汀对慢性不可预知性轻度应激（CUMS）模型大鼠抑郁样行为的作用。方法 18只SD大鼠随机分为对照组、模型组和氟西汀组。模型组和氟西汀组大鼠随机给予不可预知性轻度刺激11周，制备抑郁症模型。氟西汀组于第7~11周灌胃氟西汀（10 mg·kg^-1·d^-1），其余组大鼠灌胃1 mL生理盐水。干预结束后进行行为学检测，酶联免疫吸附试验检测脑组织中白细胞介素（IL）-1β和IL-18的含量，免疫荧光染色观察海马CA3区和皮质区中NLRP3、凋亡相关斑点样蛋白（ASC）和胱天蛋白酶1（Caspase-1）的表达情况。Western blot测定脑组织TLR4、NF-κB、NLRP3、Caspase-1和活化的Caspase-1（cleaved Caspase-1）蛋白的表达水平。结果与模型组比较，氟西汀组大鼠在旷场的运动距离及站立次数显著增多，在高架十字迷宫的运动距离增加，且在闭臂的停留时间减少，大鼠脑组织中IL-1β和IL-18含量显著降低，TLR4、NF-κB、NLRP3、ASC、Caspase-1和cleaved Caspase-1蛋白的表达降低（P<0.05）。结论氟西汀可能通过抑制TLR4/NF-κB/NLRP3炎症体信号通路，降低脑组织中炎性因子IL-1β和IL-18的水平，从而改善CUMS大鼠的抑郁样行为。

关键词: 抑郁症, 氟西汀, Toll样受体4, NF-κB, NLR家族, 热蛋白结构域包含蛋白3, 白细胞介素1β, 白细胞介素18, 半胱氨酸天冬氨酸蛋白酶1, ASC

Abstract:

Objective To investigate the effects of fluoxetine on depression-like behavior in chronic unpredictable mild stress (CUMS) model rats based on Toll-like receptor 4 (TLR4) /nuclear factor κB (NF-κB)/NOD-like receptor pyrin domain-associated protein 3 (NLRP3) inflammasome signaling pathway. Methods Eighteen SD rats were randomly divided into the control group, the model group and the fluoxetine group. Rats in the model group and the fluoxetine group were randomly given CUMS for 11 weeks to establish depression model. The fluoxetine group was treated with fluoxetine (10 mg·kg^-1·d^-1) through gastric perfusion from the 7th to the 11th week, and the other groups were given 1 mL of normal saline by gavage. Behavioral tests were conducted after the intervention. The levels of interleukin (IL)-1β and IL-18 in brain tissue were tested by enzyme-linked immunosorbent assay (ELISA). The expression of NLRP3 protein, apoptosis-associated speck-like protein (ASC) and cysteine aspartate protease 1 (Caspase-1) protein in hippocampal CA3 area and cortical area regions were observed by immunofluorescence staining. Western blot assay was used to detect the protein expression levels of TLR4, NF-κB, NLRP3, Caspase-1 and activated cysteine aspartate protease 1 (cleaved Caspase-1) in brain tissue. Results Compared with the model group, the movement distance and standing times of rats in open field test were significantly increased, the movement distance in the elevated plus maze test was increased, and the dwell time in closed arms was decreased in the fluoxetine group. The contents of IL-1β and IL-18 in brain tissue were significantly decreased, and expression levels of TLR4, NF-κB, NLRP3, ASC, Caspase-1 and cleaved Caspase-1 proteins were also down-regulated in the fluoxetine group than those of the model group (P<0.05). Conclusion Fluoxetine may improve depression-like behavior in CUMS rats by inhibiting TLR4/NF-κB/NLRP3 inflammasome signaling pathway and reducing the levels of inflammatory factors IL-1β and IL-18 in brain tissue.

Key words: depressive disorder, fluoxetine, Toll-like receptor 4, NF-kappa B, NLR family, pyrin domain-containing 3 protein, interleukin-1beta, interleukin-18, caspase 1, ASC

中图分类号:

R749.42

图/表 6

表1 Depression-like behavior changes of rats in each group

Tab. 1 各组大鼠抑郁样行为变化（n=6，$\bar{x}±s$）

组别	旷场实验		高架十字迷宫实验
组别	总运动距离（m）	站立次数	总运动距离（m）	闭臂停留时间（s）
对照组	12.14±3.89	10.67±4.84	8.24±1.79	237.98±7.32
模型组	3.75±2.25^a	3.17±3.12^a	2.21±1.18^a	284.32±7.99^a
氟西汀组	10.66±2.12^b	10.67±3.39^b	7.96±1.62^b	208.38±27.41^b
F	14.555^＊＊	7.550^＊＊	28.763^＊＊	30.342^＊＊

图1 各组CUMS大鼠脑组织IL-1β和IL-18含量比较 A：各组大鼠脑组织IL-1β含量的比较；B：各组大鼠脑组织IL-18含量的比较；a与对照组比较，b与模型组比较，P<0.05。

Fig. 1 Comparison of IL-1β and IL-18 in brain tissue of CUMS rats between the three groups

图2 各组大鼠NLRP3、ASC和Caspase-1蛋白在海马CA3区及皮质区的表达（免疫荧光染色，×200） A：NLRP3蛋白在海马CA3区及皮质区的表达；B：ASC蛋白在海马CA3区及皮质区的表达；C：Caspase-1蛋白在海马CA3区及皮质区的表达。

Fig.2 Expression levels of NLRP3, ASC and Caspase-1 proteins in hippocampal CA3 area and cortex of rats in each group (immunofluorescence staining, ?×200)

图3 各组大鼠NLRP3、ASC和Caspase-1蛋白相对表达水平的比较 A：各组大鼠脑组织中NLRP3蛋白相对表达水平的比较；B：各组大鼠脑组织中ASC蛋白相对表达水平的比较；C：各组大鼠脑组织中Caspase-1蛋白相对表达水平的比较；a与对照组比较，b与模型组比较，P<0.05。

Fig.3 Comparison of relative expression levels of NLRP3, ASC and Caspase-1 in brain tissue of rats between the three groups

图4 氟西汀对CUMS大鼠TLR4/NF-κB/NLRP3炎症体信号通路的影响 A：Western blot检测TLR4、NF-κB和cleaved Caspase-1蛋白条带图；B：Western blot检测NLRP3和Caspase-1蛋白条带图。

Fig.4 Effects of fluoxetine on TLR4/NF-κB/NLRP3 inflammasome signal pathway of CUMS rats

表2 Comparison of expression levels of TLR4, NF-κB, cleaved Caspase-1, NLRP3 and Caspase-1 between the three groups

Tab.2 各组大鼠TLR4、NF-κB、cleaved Caspase-1、NLRP3和Caspase-1蛋白表达水平比较（n=6，$\bar{x}±s$）

组别	TLR4	NF-κB	cleaved Caspase-1	NLRP3	Caspase-1
对照组	1.00±0.00	1.00±0.00	1.00±0.00	1.00±0.00	1.00±0.00
模型组	1.42±0.27^a	1.44±0.33^a	1.36±0.14^a	1.28±0.28^a	1.42±0.14^a
氟西汀组	1.12±0.21^b	0.97±0.19^b	1.06±0.17^b	0.97±0.25^b	1.07±0.17^b
F	16.689^＊＊	16.095^＊＊	28.289^＊＊	16.172^＊＊	34.212^＊＊

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氟西汀调节TLR4/NF-κB/NLRP3炎症体信号通路改善CUMS大鼠抑郁样行为

Fluoxetine improves depression-like behavior in CUMS rats by regulating TLR4/NF-κB/NLRP3 inflammasome signaling pathway

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