乌灵菌粉通过减弱小鼠海马内质网应激缓解抑郁样行为

doi:10.5246/jcps.2024.09.058

摘要/Abstract

摘要：

: 内质网(endoplasmic reticulum, ER)应激反应在人类抑郁症病例和啮齿类动物抑郁样模型中均有报道。乌灵菌粉来源于珍贵真菌Xylaria Nigripes (Kl.) Sacc, 已有研究证明其在减轻抑郁症状方面的功效。本研究的目的是探讨乌灵菌粉的抗抑郁特性及其分子机制, 特别是对内质网应激的调节作用。社交挫败应激模型是通过将一只小鼠放入有敌对小鼠的陌生环境的笼子中, 每天持续5–10分钟, 维持10天。随后, 小鼠接受口服乌灵菌粉治疗2周。采用社会趋避实验评估抑郁样行为。通过蛋白质印迹分析和实时荧光定量PCR分析社交挫败引发的内质网应激相关的蛋白和基因表达。行为学测试表明, 乌灵菌粉改善了由于社交挫败应激导致的抑郁样行为, 显著降低了社交挫败应激引起的78-kDa GRP78和PDI水平的增加。此外乌灵菌粉还降低了IRE1α和sXBP1的蛋白和mRNA的表达水平。同时乌灵菌粉显著降低了P-eIF2α、ATF4和CHOP的水平, 提高了Bcl-2与Bax的比值。以上结果提示, 乌灵菌粉可通过抑制蛋白翻译合成, 减轻海马内质网应激, 抑制细胞凋亡, 从而改善小鼠抑郁样行为。

关键词: 抑郁症, 乌灵菌粉, 内质网应激, 未折叠蛋白反应

Abstract:

The response to endoplasmic reticulum (ER) stress has been noted in both human depression cases and depression models in rodents. Wuling powder, derived from the mycelium of the esteemed fungus Xylaria Nigripes (Kl.) Sacc, has demonstrated efficacy in alleviating depressive symptoms. The purpose of this research was to explore the antidepressant properties of Wuling powder and its basic molecular effects, particularly regarding alterations in ER stress. A model of social defeat stress was created by introducing a mouse to the cage of an unfamiliar, hostile mouse for intervals of 5–10 min daily over a span of 10 d. Subsequently, the mice received oral doses of Wuling powder for 2 weeks. The social approach-avoidance assay was employed to evaluate signs of depression-like behaviors. Moreover, protein and gene expressions linked to ER stress triggered by social defeat were analyzed through Western blotting analysis and quantitative real-time PCR. The behavioral tests indicated that Wuling powder ameliorated behaviors associated with depression due to social defeat stress. Treatment with Wuling powder markedly reduced the increased levels of the 78-kDa glucose-regulated protein and protein disulfide isomerase caused by social defeat stress. It also diminished the expression of inositol-requiring enzyme 1α (IRE1α) and spliced X box-binding protein-1 (sXBP1) at the protein and mRNA levels. Furthermore, Wuling treatment notably decreased the levels of phosphorylated eukaryotic initiation factor 2 alpha kinase (P-eIF2α), activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP), simultaneously enhancing the ratio of B-cell lymphoma 2 (Bcl-2) to Bcl-2-associated X protein (Bax). These results suggested that Wuling powder could alleviate ER stress and inhibit cell apoptosis in the hippocampus by inhibiting protein translation and synthesis, thereby attenuating depressive-like behavior.

Key words: Depression, Wuling powder, Endoplasmic reticulum stress, Unfolded protein response

Supporting:

冯璐, 黄薇, 郑吉, 李冬梅, 王明洋, 刘俊雅, 范姝婕, 纪超, 杨楠, 刘雁勇. 乌灵菌粉通过减弱小鼠海马内质网应激缓解抑郁样行为[J]. 中国药学(英文版), 2024, 33(9): 783-794.

Lu Feng, Wei Huang, Ji Zheng, Dongmei Li, Mingyang Wang, Junya Liu, Shujie Fan, Chao Ji, Nan Yang, Yanyong Liu. Wuling powder alleviates depressive-like behavior by attenuating endoplasmic reticulum stress in the mouse hippocampus[J]. Journal of Chinese Pharmaceutical Sciences, 2024, 33(9): 783-794.

图/表 6

Figure 1. The diagram of social approach-avoidance test.

Figure 2. Effects of Wuling powder on social avoidance test in SDS model. Time spent in the interaction zone (A) or corner zone (B) with or without an aggressor among different groups. Data are expressed as mean ± SD and analyzed by one-way ANOVA followed by Least Significant Difference post hoc test (n = 12 per group; *P < 0.001 vs. Control group; #P < 0.001 vs. Model group).

Figure 3. Effect of Wuling powder treatment on protein GRP78 (A), PDI (B) expression in the hippocampus of SDS model mice. The intensity of each band was normalized to β-actin. All the values represented the mean ± SD and were analyzed by one-way ANOVA followed by Least Significant Difference post hoc test (n = 6 per group, *P < 0.05, ***P < 0.001 vs. Control group; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. Model group).

Figure 4. Effect of Wuling powder treatment on protein IRE1α and gene sXBP-1 expression in the hippocampus of SDS model mice. The intensity of each band was normalized to β-actin. All the values represented the mean ± SD and were analyzed by one-way ANOVA followed by the Least Significant Difference post hoc test (n = 6 per group, *P < 0.05, **P < 0.01, vs. Control group; #P < 0.05, vs. Model group).

Figure 5. Effects of Wuling powder on hippocampal protein expression level of P-eIF2α and gene expression level of ATF4 in stressed mice. Data are expressed as mean ± SD and analyzed by one-way ANOVA followed by Least Significant Difference post hoc test (n = 4; *P < 0.05 vs. Control group; #P < 0.05, ##P < 0.01 vs. Model group).

Figure 6. Effect of Wuling powder on the hippocampal gene expression level of CHOP and protein expression level of Bcl-2, Bax in stressed mice. Data are expressed as mean ± SD and analyzed by one-way ANOVA followed by Least Significant Difference post hoc test (n = 4; *P < 0.05, ***P < 0.001 vs. Control group; #P < 0.05, ##P < 0.01 vs. Model group).

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