越鞠丸治抑郁，原因竟是......

文献来源

Juanjuan Tang, Wenda Xue, Baomei Xia, et al. Involvement of normalized NMDA receptor and mTOR-related signaling in rapid antidepressant effects of Yueju and ketamine on chronically stressed mice. Sci Rep, 2015, doi: 10.1038/srep13573. (IF: 5.228)

前言

       中医经典方药越鞠丸（Yueju）出自《丹溪心法》，其为朱丹溪于800年前创制的解郁名方。越鞠丸由香附（Cyperus rotundus）、川芎（Ligusticum Chuanxiong）、栀子（Gardeian jasminoides）、苍术（Atractylodes lancea）和神曲（Massa Fermentata）五味中药组配而成。越鞠丸具有理气解郁，宽中除满之功效，常用于胸脘痞闷，腹中胀满，饮食停滞，嗳气吞酸。然而，南京中医药大学转化系统生物学与神经科学研究中心教授陈刚领衔的研究小组，于2015年首次发现越鞠丸具有独特的快速持久抗抑郁作用，并揭示了其改善抑郁脑区失常的蛋白激酶B（Akt）信号通路和持久提升脑源性神经营养因子（BDNF）的机制。

摘要

      中医经典方药越鞠丸（Yueju）具有与快速抗抑郁原型药氯胺酮（Ketamine）相似的快速持久的抗抑郁效应。本研究旨在评价越鞠丸与氯胺酮在慢性应激大鼠模型中的快速持久抗抑郁效力以及其效力发挥与前额叶N-甲基-D-天门冬氨酸（N-methyl-D-aspartate, NMDA）受体和哺乳动物的雷帕霉素靶蛋白（mammalian target of rapamycin, mTOR）相关活性变化之间的关系。慢性轻度应激（chronic mild stress, CMS）将导致糖水偏好实验（sucrose preference test, SPT）、强迫游泳实验、悬尾实验及新环境进食抑制实验的缺失，然而这些缺失可通过越鞠丸和氯胺酮的急性给药而得到不同程度的改善。越鞠丸和氯胺酮给药2小时后，SPT就得以改善，然而给予越鞠丸后其改善效果持续了6天。对于给药6天后的体重恢复，越鞠丸优于氯胺酮。CMS降低了mTOR效应体4E-BP1和p70S6K、其上游的调节子细胞外信号调节激酶（ERK）和蛋白激酶B（Akt）及下游靶点（包括突触蛋白GluR1）的磷酸化。越鞠丸或氯胺酮给药2天后，上述变化得以逆转，然而仅越鞠丸在给药6天后仍可逆转磷酸化Akt。CMS选择性、持久性的增加了NMDA受体亚单位NR1的表达，然而这一结局在氯胺酮或越鞠丸给药2天后得以逆转，然而仅越鞠丸给药6天后上述结局仍旧得以逆转。上述结果表明，NR1和Akt/mTOR信号通路是抑郁重要的治疗靶点。

ABSTRACT

      Yueju, a Traditional Chinese Medicine formula, exhibited fast-onset antidepressant responses similar to ketamine. This study focused on assessing the rapid and persistent antidepressant efficacy of Yueju and ketamine in chronically stressed mice and its association with alternations in prefrontal N-methyl-D-aspartate (NMDA) receptor and mammalian target of rapamycin (mTOR)-related activity. Chronic mild stress (CMS) led to deficits in sucrose preference test (SPT), forced swim test, tail suspension test, and novelty suppressed feeding test, which were improved differently by acute Yueju or ketamine administration. The improvement in SPT started as soon as 2 hours post Yueju and ketamine but lasted for 6 days only by Yueju. Body weight was regained by Yueju more than ketamine at post-drug administration day (PAD) 6. CMS decreased phosphorylation of the mTOR effectors 4E-BP1 and p70S6K, their upstream regulators ERK and Akt, and downstream targets including synaptic protein GluR1. Yueju or ketamine reversed these changes at PAD 2, but only Yueju reversed phosphor-Akt at PAD 6. CMS selectively and lastingly increased NMDA receptor subunit NR1 expression, which was reversed by ketamine or Yueju at PAD 2 but only by Yueju at PAD 6. These findings suggest that NR1 and Akt/mTOR signaling are important therapeutic targets for depression.

结果

图1. 氯胺酮与越鞠丸治疗CMS大鼠不同时间点的行为效应。对照大鼠给予赋形剂治疗，暴露于CMS的大鼠接受单剂量赋形剂、越鞠丸或氯胺酮治疗。(A) 给药2小时、2天及6天后糖水偏好实验结果。(B, C) 给药2小时、2天及6天后体重变化情况。(D) 给药1天及5天后悬尾实验的静止时间。(E) 给药1天及5天后强迫游泳实验的静止时间。(F) 给药2天及6天后新环境进食抑制实验期间的进食的延迟情况。(G) 给药2天及6天后新环境进食抑制实验期间总的进食量。

Figure 1. Behavioral effects at different time points after a single ketamine and Yueju treatment in CMS mice. Control animals (Con) received vehicle treatment, and animals exposed to CMS received a single administration of vehicle (Veh), Yueju (YJ) or ketamine (Ket). (A) Sucrose preference test at 2 hours, 2 days and 6 days after drug administration, respectively. (B,C) Body weight changes at 2 days and 6 days post drug administration. (D) The immobility time in the TST at 1 day and 5 days post drug administration. (E) The immobility time in the FST 1 day and 5 days after drug administration. (F) Latency to feed during the NSF test at 2 days and 6 days post drug administration. (G) Total amount of food consumed during the NSF test at 2 days and 6 days post drug administration.

图2. CMS大鼠接受氯胺酮和越鞠丸治疗2天后mTOR下游效应体活性的变化情况。(A) 4E-BP1的磷酸化水平。(B)以总的p70S6K为基准标化后磷酸化p70S6K。(C) 以总的mTOR为基准标化后的磷酸化mTOR。

Figure 2. Alternations of activation of downstream effectors of mTOR signaling in PFC of CMS mice at 2 days post ketamine and Yueju administration. (A) Phosphorylation of 4E-BP1 level. (B) Phosphorylated p70S6K (pp70S6K) normalized to total p70S6K. (C) phosphorylated mTOR (p-mTOR) normalized to total mTOR.

图3. CMS大鼠接受氯胺酮和越鞠丸治疗2天后ERK1/2和Akt的变化情况。(A) 以总的Akt为基准标化后的磷酸化Akt的光密度分析。(B) 以总的ERK1/2为基准标化后的磷酸化ERK1/2。

Figure 3. Alternations of activation of ERK1/2 and Akt in the PFC of CMS mice at 2 days post ketamine and Yueju administration. (A) Densitometric analysis of phosphorylated Akt normalized to total Akt. (B) phosphorylated ERK1/2 normalized to total ERK1/2.

图4. CMS接受氯胺酮和越鞠丸治疗后突触蛋白的变化情况。越鞠丸或氯胺酮单剂量给药后对PFC中蛋白表达水平的影响通过蛋白印迹法加以测定。(A) NR1. (B) NR2A. (C) NR2B. (D) GluR-1. (E) PSD-95. (F) synapsin-1.

Figure 4. Alternations of synaptic proteins in the PFC of mice by CMS as well as ketamine and Yueju treatment. The treatment effects on protein expression levels in the PFC were determined with western blotting at 2 days post a single administration of Yueju or ketamine. (A) NR1. (B) NR2A. (C) NR2B. (D) GluR-1. (E) PSD-95. (F) synapsin-1.

图5. CMS大鼠接受氯胺酮和越鞠丸治疗6天后mTOR信号通路及突触蛋白的变化情况。越鞠丸或氯胺酮单剂量给药后对PFC中蛋白表达水平的影响通过蛋白印迹法加以测定。 (A) NR1. (B) GluR-1. (C) 4E-BP1磷酸化水平. (D)以总的Akt为基准标化后的磷酸化Akt.

Figure 5. Alternations of synaptic proteins and mTOR signaling in the PFC of CMS mice at 6 days post ketamine and Yueju administration. The treatment effects on protein expression levels in the PFC were determined with western blotting. (A) NR1. (B) GluR-1. (C) Phosphorylation of 4E-BP1 level. (D) phosphorylated Akt normalized to total Akt.

图6. 快速抗抑郁对大鼠抑郁模型PFC中mTOR相关活性及AMPA/NMDA受体表达影响的作用模式。

Figure 6. A working model of the effects of fast-acting antidepressants on mTOR related activity and AMPA/NMDA receptor expressions in the PFC of a rodent model of depression.