茯苓多糖通过NF-<i>κ</i>B和NLRP3信号通路调节脂多糖引起的焦虑和抑郁样行为

史云静; 李玉霞

doi:10.13386/j.issn1002-0306.2022080276

茯苓多糖通过NF-κB和NLRP3信号通路调节脂多糖引起的焦虑和抑郁样行为

Poria cocos Polysaccharides Regulate Anxiety and Depression-like Behaviors Induced by Lipopolysaccharide through NF-κB and NLRP3 Signaling Pathways

摘要

摘要: 目的：探讨茯苓多糖（Poria cocos polysaccharide，PPS）对脂多糖（LPS）引起的焦虑和抑郁样行为的影响，并分析其机制。方法：BV-2细胞分成对照组、LPS组、LPS+氟西汀组、LPS+PPS（PPS分别为4、8、16 μmol/L），处理24 h，二氯二氢荧光素二氢乙酸酯（DCFH-DA）荧光探针检测细胞内活性氧（ROS）水平，Griess法检测培养上清液NO水平，酶联免疫吸附试验（ELISA）检测培养上清液中TNF-α、IL-1β水平，Western blot实验检测CD206、CD16/32、NF-κB p65水平。动物实验将小鼠分为对照组、模型组、LPS+氟西汀组、LPS+PPS低剂量组（20 mg/kg）、LPS+PPS高剂量组（80 mg/kg），每组10只，测试抑郁样行为，检测海马组织TNF-α、IL-1β、IL-18的浓度，分析海马组织CD206、CD16/32、NF-κB p65、NLRP3、ASC、Cleaved caspase-1蛋白水平。结果：与LPS组比较，4、8、16 μmol/L PPS能极显著降低ROS荧光相对强度（P<0.01），下调NO、TNF-α、IL-1β水平，降低CD16/32、NF-κB p65水平，增加CD206水平（P<0.05）；与模型组比较，LPS+PPS低剂量组、LPS+PPS高剂量组蔗糖偏爱率极显著增加（P<0.01），旷场穿越次数、修饰次数极显著上调（P<0.01），悬尾不动时间、强迫游泳不动时间极显著降低（P<0.01）；与模型组比较，LPS+PPS低剂量组、LPS+PPS高剂量组IL-1β、IL-18、TNF-α水平极显著降低（P<0.01），CD16/32、NF-κB p65水平显著下调（P<0.05），CD206显著上调（P<0.05），NLRP3、ASC、Cleaved caspase-1水平极显著降低（P<0.01）。结论：茯苓多糖能通过抑制NF-κB和NLRP3信号通路减轻LPS诱导的焦虑和抑郁样行为。

Abstract: Objective: To explore the effect of Poria cocos polysaccharides (Poria cocos polysaccharide, PPS) on anxiety and depression-like behaviors induced by lipopolysaccharide (LPS) and analyze its mechanism. Methods: BV-2 cells were divided into control group, LPS group, LPS+fluoxetine group and LPS+PPS (PPS of 4, 8, 16 μmol/L, respectively). After 24 h treatment, the level of intracellular reactive oxygen species (ROS) was detected by dichlorodihydrofluorescein dihydroacetate (DCFH-DA) fluorescence probe, the level of NO in culture supernatant was detected by griess method, and the levels of TNF-α and IL-1β in cell supernatant were detected by enzyme linked immunosorbent assay (ELISA). The levels of CD206, CD16/32 and NF-κB p65 were detected by western blot. In animal experiment, mice were divided into control group, model group, LPS+fluoxetine group, LPS+PPS low dose group (20 mg/kg) and LPS+PPS high dose group (80 mg/kg). Depression-like behavior test was performed, and the concentration of TNF-α, IL-1β and IL-18 in hippocampus was detected. The protein levels of CD206, CD16/32, NF-κB p65, NLRP3, ASC and Cleaved caspase-1 in hippocampus were analyzed. Results: Compared with LPS group, 4,8,16 μmol/L PPS significantly decreased the relative fluorescence intensity of ROS (P<0.01), decreased the levels of NO, TNF-α and IL-1β (P<0.01), decreased the levels of CD16/32 and NF-κB p65 (P<0.05), and increased the level of CD206 (P<0.05). Compared with the model group, the preference rate, crossing times and modification times in LPS+PPS low dose group and high dose LPS+PPS group significantly increased (P<0.01), while the tail suspension time and forced swimming immobility time significantly decreased (P<0.01). Compared with the model group, the levels of IL-1β, IL-18 and TNF-α in LPS+PPS low dose group and LPS+PPS high dose group significantly decreased (P<0.01), CD16/32 and NF-κB p65 level were significantly down-regulated, CD206 was significantly up-regulated, and the levels of NLRP3, ASC and Cleaved caspase-1 significantly decreased. Conclusion: Poria cocos polysaccharide can alleviate LPS-induced anxiety and depression-like behavior by inhibiting NF-κB and NLRP3 signal pathways.

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