电针对慢性不可预知温和应激小鼠模型抑郁样行为及肠道菌群的调节作用
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国家自然科学基金项目(82101594);陕西省自然科学基金(2022JQ-972)


Effect of electroacupuncture on behavior and the gut microbiome in chronic unpredictable mild stress mouse model
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    摘要:

    目的 观察电针对慢性不可预知温和应激(CUMS)模型小鼠抑郁样行为及肠道菌群组 成的影响。方法 将 40 只雄性 C57BL/6 小鼠随机分为对照组、电针组、模型组(CUMS 组)和治疗组 (CUMS+ 电针组),每组 10 只。CUMS 组和 CUMS+ 电针组均接受 CUMS 造模,造模结束后,给予对照组和 CUMS 组小鼠假刺激 7 d,给予电针组和 CUMS+ 电针组小鼠 2/15 Hz、1.0 mA 的电针刺激 7 d。最后一次 干预结束 24 h 后,收集粪便保存在 -80℃条件下并对小鼠进行糖水偏好实验、旷场实验和强迫游泳实 验。对 4 组小鼠的粪便进行 16S rDNA 测序,采用操作分类单元(OTU)信息进行 α 多样性和 β 多样性 分析、线性判别分析效应量分析(LEfSe)。采用 Spearman 相关性分析小鼠行为学指标与差异菌群富集 水平之间的相关性。结果 CUMS 组小鼠的旷场中心区域探索时间短于对照组[(22.058±4.148)s 比 (37.864±4.407)s],糖水偏好率低于对照组[(53.427±14.550)% 比(76.514±15.701)%],强迫游泳不动 时间长于对照组[(82.599±32.369)s 比(47.606±15.344)s],差异有统计学意义(P< 0.01)。CUMS+ 电针 组小鼠的旷场中心区域探索时间长于 CUMS 组[(30.604±6.060)s 比(22.058±4.148)s],糖水偏好率高于 CUMS组[(72.731±13.933)%比(53.427±14.550)%],强迫游泳不动时间短于CUMS组[(53.633±17.933)s 比(82.599±32.369)s],差异有统计学意义(P< 0.05)。CUMS 组小鼠肠道菌群的 OTU 数量、ACE 指数和 Chao 指数低于对照组,差异有统计学意义(P< 0.05);但 CUMS+ 电针组与 CUMS 组比较,差异无统计学 意义(P> 0.05)。Ruminococcaceae_UCG_002 属、蓝绿藻菌属(Lachnoclostridium)和 Rikenellaceae_RC9_ gut_group 属在 CUMS 组中富集,且与抑郁样行为呈正相关(P< 0.05);放线菌门和理研菌属、杜氏杆菌属、 Ileibacterium 属、双歧杆菌属和异杆菌属在电针和 CUMS+ 电针组中富集,且均与抑郁样行为呈负相关 (P< 0.05)。结论 电针干预可以缓解小鼠的抑郁样行为,且对其肠道菌群组成有调节作用。

    Abstract:

    Objective To investigate the impact of electroacupuncture on behavior and the gut microbiome in chronic unpredictable mild stress (CUMS) mouse model. Methods A total of 40 male C57BL/6 mice were randomly distributed into control group, electroacupuncture group, CUMS group and treatment group (CUMS + electroacupuncture), with 10 mice in each group. CUMS group and CUMS + electroacupuncture group accepted CUMS modeling. After the completion of modeling, mice in the control group and CUMS group were given sham stimulation for 7 days, and mice in the electroacupuncture group and CUMS + electroacupuncture group were given 2/15 Hz, 1.0 mA electroacupuncture stimulation for 7 days. 24 hours after the last intervention, feces were collected and stored at -80℃. Sugar preference test, open field test and forced swimming test were conducted on mice. 16S rDNA sequencing was performed on the feces of four groups of mice. α diversity,β diversity analysis and Linear Discriminant analysis Effect Size (LEfSe) were carried out by Operational Taxonomic Units (OTU) information. Spearman correlation analysis of the correlation between the behavioral indicators of mice and the enrichment levels of different bacterial populations. Results The time of exploring the open field center area of CUMS group mice was shorter than that of the control group [(22.058±4.148) s vs (37.864±4.407) s], the sugar preference rate was lower than that of the control group [(53.427±14.550)% vs (76.514±15.701)%], and the forced swimming immobility time was longer than that of the control group [(82.599±32.369) s vs (47.606±15.344) s], the difference was statistically significant (P< 0.01). The time of exploring the open field center area of CUMS + electroacupuncture group mice was longer than that of the CUMS group [(30.604±6.060) s vs (22.058±4.148) s], the sugar preference rate was higher than that of the CUMS group [(72.731±13.933)% vs (53.427±14.550)%], and the forced swimming immobility time was shorter than that of the CUMS group [(53.633±17.933) s vs (82.599±32.369) s], the difference was statistically significant (P < 0.05). The number of OTU, ACE index and Chao index of intestinal flora in CUMS group were lower than those in the control group, and the difference was statistically significant (P < 0.05), but there was no statistical significance in the difference between CUMS + electroacupuncture group and CUMS group (P>0.05). Ruminococcaceae_UCG-002, Lachnoclostridium and Rikenella ceae_RC9_gut_group were abundant in the CUMS group, which were positively correlated with depressive-like behavior (P< 0.05). Phylum Actinobacteria and genus Rikenella, Dubosiella, Ileibacterium, Bifidobacterium and Allobaculum were enriched in the electroacupuncture group and CUMS + electroacupuncture groups, all of which were negatively correlated with depressive-like behaviors. Conclusions Electroacupuncture treatment can ameliorate depressive-like behavior and regulate the composition of gut microbiota in CUMS-treated mice.

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白洁,薛姗姗,周翠红,薛芬,石青青,何宏.电针对慢性不可预知温和应激小鼠模型抑郁样行为及肠道菌群的调节作用[J].神经疾病与精神卫生,2022,22(11):
DOI :10.3969/j. issn.1009-6574.2022.11.008.

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  • 在线发布日期: 2022-12-22