Effect of <italic style="font-style: italic">Ganyu Pixu Formula</italic> （肝郁脾虚方）on Regulation of GABAergic Signaling System and Intestinal Flora in Colon Tissue of Depressed and Insomniac Model Rats

CHEN Shupeng; TANG Nana; WANG Simeng; WANG Zhuhua; XU Yuanyuan; MIAO Huihui; HUANG Junshan

doi:10.13288/j.11-2166/r.2023.12.014

您当前的位置：

首页 >

文章列表页 >

Effect of Ganyu Pixu Formula （肝郁脾虚方）on Regulation of GABAergic Signaling System and Intestinal Flora in Colon Tissue of Depressed and Insomniac Model Rats

更新时间：2023-06-19

- Effect of Ganyu Pixu Formula （肝郁脾虚方）on Regulation of GABAergic Signaling System and Intestinal Flora in Colon Tissue of Depressed and Insomniac Model Rats
- Journal of Traditional Chinese Medicine Vol. 64, Issue 12, Pages: 1273-1281(2023)
- 作者机构：
  
  1.江西中医药大学临床医学院，江西省南昌市新建区梅岭大道1688号，330004
  2.江西中医药大学附属医院
  3.福建省中医药科学院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2023.12.014
  CLC：
- Received：04 October 2022，
  
  Revised：2023-02-16，
  
  Published：17 June 2023
- 稿件说明：
移动端阅览
陈抒鹏,唐娜娜,王思梦等.肝郁脾虚方对抑郁失眠模型大鼠结肠组织GABA能信号系统及肠道菌群的影响[J].中医杂志,2023,64(12):1273-1281.

CHEN Shupeng,TANG Nana,WANG Simeng,et al.Effect of Ganyu Pixu Formula （肝郁脾虚方）on Regulation of GABAergic Signaling System and Intestinal Flora in Colon Tissue of Depressed and Insomniac Model Rats[J].Journal of Traditional Chinese Medicine,2023,64(12):1273-1281.
陈抒鹏,唐娜娜,王思梦等.肝郁脾虚方对抑郁失眠模型大鼠结肠组织GABA能信号系统及肠道菌群的影响[J].中医杂志,2023,64(12):1273-1281. DOI： 10.13288/j.11-2166/r.2023.12.014.

CHEN Shupeng,TANG Nana,WANG Simeng,et al.Effect of Ganyu Pixu Formula （肝郁脾虚方）on Regulation of GABAergic Signaling System and Intestinal Flora in Colon Tissue of Depressed and Insomniac Model Rats[J].Journal of Traditional Chinese Medicine,2023,64(12):1273-1281. DOI： 10.13288/j.11-2166/r.2023.12.014.

摘要

目的

探究肝郁脾虚方治疗抑郁共病失眠的可能作用机制。

方法

24只SD大鼠随机分为空白组，模型组及肝郁脾虚方低、高剂量组，每组6只。除空白组外，其余各组均采用慢性不可预知温和应激方法（CUMS）干预21天构建抑郁模型，再注射4-氯-DL-苯丙氨酸（PCPA）3天构建抑郁失眠大鼠模型。造模后肝郁脾虚方低、高剂量组分别予以肝郁脾虚方8、32 g/（kg·d）灌胃，空白组及模型组以蒸馏水20 ml/（kg·d）灌胃，共灌胃14天。采用实时荧光定量PCR检测大鼠结肠组织γ-氨基丁酸A型受体（GABAA）、γ-氨基丁酸B型受体（GABAB）、γ-氨基丁酸转运蛋白（GAT）、谷氨酸脱羧酶（GAD） mRNA表达，Western blot法检测大鼠结肠组织中GABAA、GABAB、GAT、GAD蛋白表达。通过16SRNA基因测序技术分析大鼠肠道菌群物种多样性及物种组成。Pearson相关性分析γ-氨基丁酸（GABA）能信号系统与优势菌门的相关性。

结果

与空白组比较，模型组大鼠结肠组织GABAA、GABAB蛋白及mRNA表达降低，GAD蛋白及mRNA、GAT蛋白表达升高（

＜0.05）。与模型组比较，肝郁脾虚方高、低剂量组GABAA蛋白及mRNA表达升高，GABAB mRNA表达升高、蛋白表达下降，GAD mRNA表达降低，GAT蛋白表达下降（

＜0.05）。与肝郁脾虚方低剂量组比较，肝郁脾虚方高剂量组GABAA、GAD mRNA表达升高，GABAB蛋白表达降低（

＜0.05）。各组均未检测到GAT mRNA表达。Alpha多样性结果显示，与模型组比较，肝郁脾虚方低、高剂量组Shannoneve指数升高，Simpson指数降低（

＜0.05）。Beta多样性结果显示，肝郁脾虚方低剂量组菌群结构更接近空白组。物种组成结果显示，各组均存在的优势菌门为厚壁菌门、拟杆菌门。Pearson相关分析显示，厚壁菌门丰度与GAD mRNA表达呈负相关（

=-0.65，

=0.02），与GABAA蛋白表达呈负相关（

=-0.80，

=0.02），拟杆菌门丰度与GAD mRNA（

=0.79，

=0.02）、GABAB mRNA呈正相关（

=0.69，

=0.01），与GABAA蛋白表达呈正相关（

=0.63，

=0.02）。

结论

肝郁脾虚方治疗抑郁共病失眠可能是通过调节肠道微生物分布，同时上调GABAA蛋白，下调GABAB、GAT蛋白实现的，且低剂量较高剂量更佳。

Abstract

Objective

To explore the possible mechanism of

Ganyu Pixu Formula

（肝郁脾虚方， GPF） in treatment of depression-comorbid insomnia （DCI）.

Methods

Twenty four SD rats were selected and randomly divided into blank group， model group， low- and high-dose GPF groups， with six rats in each group. Except for blank group， the depression and insomnia models were made by chronic unpredictable mild stress （CUMS） intervention for 21 days combined with 4-chloro-DL-phenylalanine （PCPA） injection for three days. After successful modeling， 8 g/（kg·d） and 32 g/（kg·d） of GPF were administered by gavage in the low- and high-dose GPF groups， respectively， while 20 ml/（kg·d） of distilled water was given by gavage to the blank and model groups， all for 14 days. The mRNA expression of γ-aminobutyric acid type A receptor （GABAA）， γ-aminobutyric acid type B receptor （GABAB）， γ-aminobutyric acid transporter （GAT） and glutamate decarboxylase （GAD） in rat colon tissue was detected by real-time fluorescent PCR， and the protein expression of GABAA， GABAB， GAT and GAD in rat colon tissue was detected by Western blot. The species diversity and species composition of rat intestinal flora were analyzed by 16SRNA gene sequencing technology. Pearson correlation analysis was used to analyze the correlation between GABAergic signal system and intestinal flora.

Results

Compared to those in the blank group， the protein and mRNA expression of GABAA and GABAB in the colon tissue of rats in the model group decreased， while GAD protein and mRNA expression and GAT protein expression increased （

＜0.05）. Compared to those in the model group， expression of GABAA protein， GABAA mRNA and GABAB mRNA expression in the high- and low-dose GPF groups increased， while expression of GABAB protein， GAT protein and GAD mRNA decreased （

＜0.05）. The high-dose GPF group had higher GABAA and GAD mRNA expression and lower GABAB protein expression than the low-dose group （

＜0.05）. The expression of GAT mRNA was not detected in the four groups. Alpha diversity showed that the high- and low-dose GPF groups had higher Shannoneve index and lower Simpson index compared to the model group （

＜0.05）. Beta diversity showed that the intestinal flora structure of low-dose GPF group was closer to that of the blank group. The composition of species showed that the dominant bacterial phyla in all groups were Firmicutes and Bacteroidetes. Pearson correlation analysis showed that Firmicutes abundance was negatively correlated to GAD mRNA expression （

= -0.65，

= 0.02）， and GABAA protein expression （

= -0.80，

= 0.02）. The abundance of Bacteroidetes was positively correlated to GAD mRNA （

= 0.79，

= 0.02）， GABAB mRNA （

= 0.69，

= 0.01）， and GABAA protein expression （

= 0.63，

= 0.02）.

Conclusion

GPF may treat DCI through regulating intestinal flora distribution， up-regulating GABAA protein， and down-regulating GABAB and GAT proteins， and the lower dose has better effects.

关键词

Keywords

references

陆林，马辛 . 精神病学［M］. 北京：人民卫生出版社， 2020 ： 370 .

SCHMITT K ， HOLSBOER-TRACHSLER E ， ECKERT A . BDNF in sleep， insomnia， and sleep deprivation ［J］. Ann Med ， 2016 ， 48 （ 1/2 ）： 42 - 51 .

RIEMANN D ， KRONE LB ， WULFF K ， et al . Sleep， insomnia， and depression ［J］. Neuropsychopharmacology ， 2020 ， 45 （ 1 ）： 74 - 89 .

李姿蓉，韩远山，王宇红，等 . 从抑郁共病失眠机制探寻抑郁症新药研发的方向［J］. 湖南中医药大学学报， 2019 ， 39 （ 10 ）： 1279 - 1284 .

LI Y ， HAO Y ， FAN F ， et al . The role of microbiome in insomnia， circadian disturbance and depression ［J］. Front Psychiatry ， 2018 ， 9 ： 669 . doi： 10.3389/fpsyt. 2018. 00669 http://dx.doi.org/10.3389/fpsyt.2018.00669 .

KIM S ， JO K ， HONG KB ， et al . GABA and l-theanine mixture decreases sleep latency and improves NREM sleep ［J］. Pharm Biol ， 2019 ， 57 （ 1 ）： 65 - 73 .

BRAVO JA ， FORSYTHE P ， CHEW MV ， et al . Ingestion of Lactobacillus strain regulates emotional behavior and central GABA receptor expression in a mouse via the vagus nerve ［J］. Proc Natl Acad Sci U S A ， 2011 ， 108 （ 38 ）： 16050 - 16055 .

LYTE M . Microbial endocrinology： Host-microbiota neuroendocrine interactions influencing brain and behavior ［J］. Gut Microbes ， 2014 ， 5 （ 3 ）： 381 - 389 .

邓晶晶，黄俊山，詹夏菲，等 . 黄俊山教授应用甘松治疗失眠经验［J］. 时珍国医国药， 2018 ， 29 （ 4 ）： 970 - 971 .

张瑜，王秀峰，曾雪爱，等 . 肝郁脾虚方对失眠大鼠血浆和下丘脑胆囊收缩素-8的影响［J］. 世界睡眠医学杂志， 2021 ， 8 （ 1 ）： 38 - 40 .

LACH G ， SCHELLEKENS H ， DINAN TG ， et al . Anxiety， depression， and the microbiome： a role for gut peptides ［J］. Neurotherapeutics ， 2018 ， 15 （ 1 ）： 36 - 59 .

LANZA M ， MAKOVEC F . Cholecystokinin （CCK） increases GABA release in the rat anterior nucleus accumbens via CCK（B） receptors located on glutamatergic interneurons ［J］. Naunyn Schmiedebergs Arch Pharmacol ， 2000 ， 361 （ 1 ）： 33 - 38 .

徐向青，徐向东，宋婷，等 . 药对刺五加栀子对慢性应激抑郁模型大鼠大脑PKC-IP_3信号通路的影响［J］. 中医杂志， 2017 ， 58 （ 5 ）： 414 - 417 .

李杉杉，刘新民，王琼 . 基于文献数据库的慢性不可预见性温和应激诱发抑郁症动物模型研究［J］. 中国比较医学杂志， 2021 ， 31 （ 4 ）： 77 - 83 .

肖成荣，马增春，李海静，等 . PCPA失眠大鼠模型的制作及其机制［J］. 毒理学杂志， 2007 ， 21 （ 4 ）： 326 .

李仪奎 . 中药药理实验方法学［M］. 上海：上海科学技术出版社， 2006 ： 25 .

MURPHY MJ ， PETERSON MJ . Sleep disturbances in depression ［J］. Sleep Med Clin ， 2015 ， 10 （ 1 ）： 17 - 23 .

程金湘，张丽萍，宿长军 . 慢性失眠障碍患者长期使用苯二氮䓬类药物停药的研究进展［J］. 中国全科医学， 2022 ， 25 （ 27 ）： 3347 - 3351 .

宿长军，刘煜，李柱一，等 . 黛力新与多塞平联合治疗原发性失眠［J］. 第四军医大学学报， 2005 ， 26 （ 5 ）： 96 - 98 .

WICHNIAK A ， WIERZBICKA AE ， JAREMA M . Treatment of insomnia-effect of trazodone and hypnotics on sleep ［J］. Psychiatr Pol ， 2021 ， 55 （ 4 ）： 743 - 755 .

TRINCAVELLI ML ， DA PE ， DANIELE S ， et al . The GABAA-BZR complex as target for the development of anxiolytic drugs ［J］. Curr Top Med Chem ， 2012 ， 12 （ 4 ）： 254 - 269 .

MÖHLER H . The GABA system in anxiety and depression and its therapeutic potential ［J］. Neuropharmacology ， 2012 ， 62 （ 1 ）： 42 - 53 .

BHAGAT K ， SINGH JV ， PAGARE PP ， et al . Rational approaches for the design of various GABA modulators and their clinical progression ［J］. Mol Divers ， 2021 ， 25 （ 1 ）： 551 - 601 .

DIJK DJ . Slow-wave sleep deficiency and enhancement： implications for insomnia and its management ［J］. World J Biol Psychiatry ， 2010 ， 11 （ Suppl 1 ）： 22 - 28 .

何婷 . 慢性应激性抑郁发生与海马Glu和GABA水平变化及其受体机制［D］. 西安：陕西师范大学， 2011 .

刘世钰，何丹，赵亮，等 . 养心氏片抗抑郁作用的蛋白组学研究［J］. 中南药学， 2018 ， 16 （ 5 ）： 593 - 601 .

YUNES RA ， POLUEKTOVA EU ， DYACHKOVA MS ， et al . GABA production and structure of gadB/gadC genes in Lactobacillus and Bifidobacterium strains from human microbiota ［J］. Anaerobe ， 2016 ， 42 ： 197 - 204 . doi： 10.1016/j.anaerobe.2016.10.011 http://dx.doi.org/10.1016/j.anaerobe.2016.10.011 .

STRANDWITZ P ， KIM KH ， TEREKHOVA D ， et al . GABA-modulating bacteria of the human gut microbiota ［J］. Nat Microbiol ， 2019 ， 4 （ 3 ）： 396 - 403 .

LIANG L ， ZHOU H ， ZHANG S ， et al . Effects of gut microbiota disturbance induced in early life on the expression of extrasynaptic GABA-A receptor α5 and δ subunits in the hippocampus of adult rats ［J］. Brain Res Bull ， 2017 ， 135 ： 113 - 119 . doi： 10.1016/j.brainresbull.2017.09.014 http://dx.doi.org/10.1016/j.brainresbull.2017.09.014 .

LI H ， XIAO J ， LI X ， et al . Low cerebral exposure cannot hinder the neuroprotective effects of panax notoginsenosides ［J］. Drug Metab Dispos ， 2018 ， 46 （ 1 ）： 53 - 65 .

LIN P ， DING B ， FENG C ， et al . Prevotella and Klebsiella proportions in fecal microbial communities are potential characteristic parameters for patients with major depressive disorder ［J］. J Affect Disord ， 2017 ， 207 ： 300 - 304 . doi： 10.1016/j.jad.2016.09.051 http://dx.doi.org/10.1016/j.jad.2016.09.051 .

查看原文

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Relationship between Five-Pattern Personality and Polysomnography of Depression Patients

Theoretical Basis and Application Prospects of Aromatherapy in Traditional Chinese Medicine for Treating Depression

Treatment of Chronic Insomnia Based on the Theory that "Spleen Fuctions as Wei Qi"：An Approach from the "Gut Microbiota-Circadian Rhythm-Ying-Wei Circulation" Perspective

ZHANG Zhiyuan's Experience in Treating Perimenopause-Related Insomnia with the Method of Regulating the Heart and Governing the Spirit

Effect of Hewei Anshen Formula （和胃安神方） on the Glu/GABA-Gln Cycle in the Suprachiasmatic Nucleus of Insomnia Model Rats

Related Author

LV Xueyu

LIU Yanjiao

LIN Yingna

LV Menghan

WANG Weidong

JIANG Molin

SHEN Hui

LI Yang

Related Institution

Guang’ anmen Hospital,China Academy of Chinese Medical Sciences

Beijing Hospital of Traditional Chinese Medicine，Capital Medical University

The Second Clinical Medical College of Guangzhou University of Chinese Medicine

The Second Affiliated Hospital of Guangzhou University of Chinese Medicine

First Clinical School of Shandong University of Traditional Chinese Medicine

Address：No.16, Nanxiao street, Dongzhimen, Dongcheng District, Beijing Postal code：100700
Tel：010-64035632 Email：jtcmcn@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010102007558号
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰

Effect of Ganyu Pixu Formula （肝郁脾虚方）on Regulation of GABAergic Signaling System and Intestinal Flora in Colon Tissue of Depressed and Insomniac Model Rats

DOI：10.13288/j.11-2166/r.2023.12.014

摘要

Abstract

关键词

Keywords

references