酸枣仁汤对氯苯丙氨酸诱导失眠大鼠模型治疗机制的研究

doi:10.3976/j.issn.1002-4026.2021.06.005

摘要/Abstract

摘要：

为探究经典名方酸枣仁汤对氯苯丙氨酸(PCPA)诱导失眠大鼠的作用机制,采用8周龄的雄雌性SD大鼠各20只,每10只大鼠(雌雄各半)为一组,将40只大鼠随机分成酸枣仁汤组、地西泮组、模型组和空白组,利用腹腔注射PCPA混悬液的方法构建失眠大鼠模型,通过旷场实验和高架十字迷宫实验评价造模后大鼠睡眠行为、情绪的改变。给药两周后,通过血清学检测单胺类神经递质、氨基酸类神经递质、炎性细胞的变化。高架十字迷宫实验结果显示,与空白组相比,PCPA诱导的失眠大鼠开臂停留时间百分比(t_oc)和进入开臂次数百分比(N_oc)均下降,其中t_oc显著下降,表明模型构建成功。与模型组相比,酸枣仁汤组5-羟色胺(5-HT)、γ-氨基丁酸(GABA)含量显著升高,而去甲肾上腺素(NE)含量升高不明显,多巴胺(DA)、谷氨酸(Glu)含量显著降低;酸枣仁汤组促炎性细胞因子白介素1β(IL-1β)含量显著降低,抗炎性细胞因子白介素10(IL-10)含量显著升高。与地西泮组相比,酸枣仁汤组DA含量降低更加显著,地西泮组肿瘤坏死因子(TNF-α)的含量比酸枣仁组降低更加显著,而5-HT、GABA、NE、Glu、IL-1β、IL-10含量变化趋势相同,且无显著差异。研究认为酸枣仁汤通过增加抑制性神经递质,降低兴奋性神经递质,恢复促炎性和抗炎性细胞因子水平,从而缓解失眠症状,为酸枣仁汤治疗失眠的功效机制及遣药组方提供了科学依据。

关键词: 酸枣仁汤, 失眠, 大鼠模型, 旷场实验, 高架十字迷宫实验, 神经递质, 炎性细胞因子

Abstract:

The therapeutic effects and mechanism of action of Suanzaoren decoction on p-chlorophenylalanine (PCPA)-induced insomnia in rats were investigated. We randomly divided 20 male and 20 female 8-week-old Sprague-Dawley rats into four groups of ten rats (five male and five female rats) each as follows: Suanzaoren decoction group, diazepam group, model group, and blank group. We established a rat model of insomnia by the intraperitoneal injection of PCPA suspension and evaluated the changes in the sleep behavior and mood of rats after an open-field experiment and an elevated plus-maze experiment. After two weeks of treatment, rat serum samples were collected to analyze the changes in the levels of monoamine neurotransmitters, amino acid neurotransmitters, and inflammatory factors. According to the results of the elevated plus-maze experiment, compared with the blank group, rats with PCPA-induced insomnia had a significant decrease in the percentage of open arm residence time (t_oc) and a decrease in the percentage of open arm entry time (N_oc), which indicated that the model was constructed successfully. Compared to the model group, the levels of 5-hydroxytryptamine (5-HT) and γ-aminobutyric acid (GABA) in the Suanzaoren decoction group increased significantly, whereas the increase in norepinephrine (NE) level was not significant. In contrast, dopamine (DA) and glutamic acid (Glu) levels decreased significantly. Compared to the model group, pro-inflammatory cytokine interleukin (IL)-1β level was significantly decreased, and anti-inflammatory cytokine IL-10 level was significantly increased in Suanzaoren decoction group. Compared to the diazepam group, the decrease in DA level in Suanzaoren decoction group was more significant, whereas the level of tumor necrosis factor α (TNF-α) in diazepam group reduced more significantly than that in Suanzaoren group. However, the levels of 5-HT, GABA, NE, Glu, IL-1β, and IL-10 in these two groups showed similar trends with no statistically significant differences. The study suggested that Suanzaoren decoction relieved insomnia by increasing the levels of inhibitory neurotransmitters, reducing those of excitatory neurotransmitters, and restoring those of pro- and anti-inflammatory cytokines. This study will serve as a scientific basis for demonstrating the efficacy and validating the prescription of Suanzaoren decoction in treating insomnia.

Key words: Suanzaoren decoction, insomnia, rat model, open field experiment, elevated plus-maze experiment, neurotransmitter, inflammatory cytokines

中图分类号:

R285.5

刘昊, 韩昌胜, 迟显苏, 杨振霄, 王祥宇, 王新, 马柯. 酸枣仁汤对氯苯丙氨酸诱导失眠大鼠模型治疗机制的研究[J]. 山东科学, 2021, 34(6): 32-40.

LIU Hao, HAN Chang-sheng, CHI Xian-su, YANG Zhen-xiao, WANG Xiang-yu, WANG Xin, MA Ke. Therapeutic effects and mechanism of action of Suanzaoren decoction, a classical prescription, on p-chlorophenylalanine-induced insomnia in rats[J]. Shandong Science, 2021, 34(6): 32-40.

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组别	总距离/cm	中央区进入次数/次	中央区时间/s
空白组	1 923.36±266.16	3.00±1.41	527.37±256.42
模型组	1 260.30±136.15^*	1.67±0.35^***	381.61±83.01^***

组别	开臂停留时间百分比(t_oc)	进入开臂次数百分比(N_oc)
空白组	0.076±0.038	0.119±0.018
模型组	0.056±0.012^***	0.106±0.024^*

组别	ρ(5-HT)/(ng·L^-1)	ρ(DA)/(pg·mL^-1)	ρ(NE)/(ng·L^-1)
空白组	325.02±58.42	65.77±6.76	101.47±11.54
模型组	243.85±52.72^*	74.34±8.55^*	145.79±20.19^*
地西泮组	270.74±11.79^※	81.88±3.90^※	157.71±22.18^※
酸枣仁汤组	330.68±44.60^※	61.31±3.82^※※	153.81±22.77^※※※

组别	c(Glu)/(μmol·L^-1)	c(GABA)/(μmol·L^-1)
空白组	10.13±1.55	1 879.46±215.85
模型组	12.55±1.05^*	1 302.80±318.80^**
地西泮组	7.63±1.17^※※	1 932.75±170.51^※※
酸枣仁汤组	9.31±1.81^※※	1 721.13±181.81^※※

组别	ρ(TNF-α)/(ng·L^-1)	ρ(IL-1β)/(ng·L^-1)	ρ(IL-10)/(ng·L^-1)
空白组	224.65±27.72	26.83±2.89	101.60±11.99
模型组	302.32±67.31^**	33.04±3.31^**	59.62±10.28^**
地西泮组	230.00±32.07^※※	26.15±5.59^※※	99.35±15.16^※※
酸枣仁汤组	316.72±30.41^※※※	24.67±2.32^※※	90.13±16.13^※※