Effects of <italic style="font-style: italic">Bushen Huoxue Formula</italic> （补肾活血方） on TLR/NF-κB Pathway and Intestinal Flora in Ileum Tissue of Parkinson's Disease Model Mice

QI Xiaorong; HAO Feiran; TANG Xianglin; LI Fagen; WANG Yujia; WANG Liang; SHEN Yingfan; YANG Minghui; LI Min

doi:10.13288/j.11-2166/r.2024.10.011

您当前的位置：

首页 >

文章列表页 >

Effects of Bushen Huoxue Formula （补肾活血方） on TLR/NF-κB Pathway and Intestinal Flora in Ileum Tissue of Parkinson's Disease Model Mice

更新时间：2024-05-13

- Effects of Bushen Huoxue Formula （补肾活血方） on TLR/NF-κB Pathway and Intestinal Flora in Ileum Tissue of Parkinson's Disease Model Mice
- Journal of Traditional Chinese Medicine Vol. 65, Issue 10, Pages: 1038-1045(2024)
- 作者机构：
  
  1.中国人民解放军总医院第六医学中心中医医学部，北京市海淀区阜成路6号，100048
  2.中国人民解放军军事科学院军事医学研究院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2024.10.011
  CLC：
- Received：21 December 2023，
  
  Revised：2024-02-29，
  
  Published：17 May 2024
- 稿件说明：
移动端阅览
齐小荣,郝斐然,汤响林等.补肾活血方对帕金森病模型小鼠回肠组织TLR/NF-κB通路及肠道菌群的影响[J].中医杂志,2024,65(10):1038-1045.

QI Xiaorong,HAO Feiran,TANG Xianglin,et al.Effects of Bushen Huoxue Formula （补肾活血方） on TLR/NF-κB Pathway and Intestinal Flora in Ileum Tissue of Parkinson's Disease Model Mice[J].Journal of Traditional Chinese Medicine,2024,65(10):1038-1045.
齐小荣,郝斐然,汤响林等.补肾活血方对帕金森病模型小鼠回肠组织TLR/NF-κB通路及肠道菌群的影响[J].中医杂志,2024,65(10):1038-1045. DOI： 10.13288/j.11-2166/r.2024.10.011.

QI Xiaorong,HAO Feiran,TANG Xianglin,et al.Effects of Bushen Huoxue Formula （补肾活血方） on TLR/NF-κB Pathway and Intestinal Flora in Ileum Tissue of Parkinson's Disease Model Mice[J].Journal of Traditional Chinese Medicine,2024,65(10):1038-1045. DOI： 10.13288/j.11-2166/r.2024.10.011.

摘要

目的

从肠道菌群探讨补肾活血方治疗帕金森病（PD）的可能作用机制。

方法

将72只雄性C57/BL6J小鼠随机分为空白组、模型组、美多芭组及补肾活血方低、中、高剂量组，每组12只。空白组小鼠腹腔注射10 ml/kg生理盐水，其余各组小鼠采用腹腔注射浓度为3 mg/ml的1-甲基-4-苯基-1

6-四氢吡啶（MPTP）30 mg/kg诱导PD小鼠模型，均每天1次，连续7天。造模成功后，补肾活血方低、中、高剂量组分别给予补肾活血方7.5、15、30 g/（kg·d）灌胃，美多芭组给予多美多芭片112.5 mg/（kg·d）灌胃，空白组和模型组予15 ml/（kg·d）蒸馏水灌胃，均每天灌胃1次，连续14天。采用爬杆实验、转棒实验、抓力实验和负重游泳实验评估小鼠行为学指标；16S rRNA高通量测序技术分析各组小鼠肠道菌群改变；Western Blot法测定小鼠回肠组织Toll样受体（TLR）/核因子κB（NF-κB）通路炎症因子［包括Toll样受体2（TLR2）、Toll样受体4（TLR4）、NF-κB、肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）、白细胞介素17（IL-17）］蛋白表达水平。

结果

与空白组比较，模型组小鼠爬杆触底时间延长，抓力减小，转棒持续时间和游泳持续时间缩短，回肠组织中TLR2、TLR4、NF-κB、TNF-α、IL-6、IL-17蛋白表达均升高（

＜0.01）。与模型组比较，美多芭组和补肾活血方低、中、高剂量组爬杆触底时间缩短，抓力增加，美多芭组和补肾活血方高剂量组转棒持续时间延长，回肠组织TLR2、TLR4、NF-κB、TNF-α、IL-6、IL-17蛋白表达均降低，而仅有补肾活血方高剂量组游泳持续时间延长（

＜0.05或

＜0.01）。与补肾活血方低剂量组比较，补肾活血方中、高剂量组爬杆触底时间缩短（

＜0.05或

＜0.01）；补肾活血方高剂量组抓力增加，TLR2、TLR4、IL-17蛋白表达降低（

＜0.05或

＜0.01）。肠道菌群结果显示，与空白组比较，模型组Dominance指数、Pielou_e指数、Shannon指数、Simpson指数差异均具有统计学意义（

＜0.05或

＜0.01）；与模型组比较，美多芭组Shannon指数、Chao1指数、Observed_otus指数，以及补肾活血方高剂量组Chao1指数、Observed_otus指数、Dominance指数、Pielou_e指数、Shannon指数、Simpson指数差异均具有统计学意义（

＜0.05或

＜0.01）。在门水平上，各组差异具有统计学意义的菌门相对丰度类别包括变形菌门、拟杆菌门、厚壁菌门（

＜0.05或

＜0.01）；在属水平上，各组差异具有统计学意义的菌属相对丰度类别包括Muribaculaceae、阿克曼菌属、幽门螺杆菌（

＜0.05或

＜0.01）。

结论

补肾活血方治疗PD的可能作用机制是通过重构紊乱的肠道菌群结构，减轻炎症反应进而达到改善PD运动功能的目的。

Abstract

Objective

To explore the possible mechanism of

Bushen Huoxue Formula

（补肾活血方， BHF） in the treatment of Parkinson's dise

ase （PD） from the the perspective of intestinal flora.

Methods

Seventy-two male C57/BL6J mice were randomly divided into blank group， model group， Madopar group and low-， medium- and high-dose BHF groups， with 12 mice in each group. The mice in the blank group were intraperitoneally injected with 10 ml/kg of normal saline， and those in the other groups were intraperitoneally injected with 30 mg/kg of 1-methyl-4-phenyl-1

6-tetrahydropyridine （MPTP） at a concentration of 3 mg/ml to induce PD mice model， both once a day for 7 consecutive days. After successful modeling， the low-， medium-， and high-dose BHF groups were given 7.5， 15， and 30 g/（kg·d） of BHF by gavage， respectively， while the Madopar group was given 112.5 mg/（kg ·d） of Domedopar tablets by gavage， and the blank group and the model group were given 15 ml/（kg·d） of distilled water， all once a day for 14 consecutive days. The rod climbing test， rotating rod test， grip strength test and weight-bearing swimming test were used to evaluate the behavioral indicators of mice. Western blotting was used to measure the protein expression levels of Toll-like receptor （TLR）/nuclear factor kappa B （NF-κB） pathway inflammatory factors in the mouse ileum， including Toll-like receptor 2 （TLR2）， Toll-like receptor 4 （TLR4）， NF-κB， tumor necrosis factor α （TNF-α）， interleukin 6 （IL-6）， and interleukin 17 （IL- 17）. 16S rRNA high-throughput sequencing was used to analyze changes in mouse intestinal flora.

Results

Compared to those in the blank group， the mice in the model group had longer bottoming time when climbing the pole， reduced grip strength， shortened rotary pole duration and swimming duration， and increased protein expression levels of TLR2， TLR4， NF-κB， TNF-α， and IL-6 in the ileal tissue （

＜0.01）. Compared to the model group， the Madopar group and the low-， medium- and high-dose BHF groups had shortened bottoming time of the climbing pole and increased grip strength； the Madopar gr

oup and the high-dose BHF group had prolonged rotary pole duration， and reduced protein expressions of TLR2， TLR4， NF-κB， TNF-α， IL-6， and IL-17 levels； and only the high-dose BHF group had prolonged swimming duration （

＜0.05 or

＜0.01）. Compared to those in the low-dose BHF group， the bottoming time of the climbing pole were shorter in the moderate- and high-dose groups （

＜0.05 or

＜0.01）， and the grip strength increased while the protein expression levels of TLR2， TLR4 and IL-17 decreased in the high-dose group （

＜0.05 or

＜0.01）. The intestinal flora results showed significant differences between the blank group and the model group in the Dominance index， Pielou_e index， Shannon index， and Simpson index （

＜0.05 or

＜0.01）. Compared to those of the model group， the Shannon index， Chao1 index， and Observed_otus index of the Madopar group， as well as the Chao1 index， Observed_otus index， Dominance index， Pielou_e index， Shannon index， and Simpson index of the high-dose BHF group all showed significantly statistical differences （

＜0.05 or

＜0.01）. At the phylum level， the relative abundance categories of bacterial phyla with statistically significant differences in each group included Proteobacteria， Bacteroidetes， and Firmicutes （

＜0.05 or

＜0.01）. At the genus level， the relative abundance categories of bacterial genera with statistically significant diffe-rences among each group included Muribaculaceae， Akkermansia， and Helicobacter pylori （

＜0.05 or

＜0.01）.

Conclusion

The possible mechanism of BHF in treating PD may be to reconstruct the disordered intestinal flora structure and improve the inflammatory response.

关键词

Keywords

references

LU QB ， ZHU ZF ， ZHANG HP ， et al . Lewy pathological study on α-synuclein in gastrointestinal tissues of prodromal Parkinson's disease ［J］. Eur Rev Med Pharmacol Sci ， 2017 ， 21 （ 7 ）： 1514 - 1521 .

HOLMQVIST S ， CHUTNA O ， BOUSSET L ， et al . Direct evidence of Parkinson pathology spread from the gastrointestinal tract to the brain in rats ［J］. Acta Neuropathol ， 2014 ， 128 （ 6 ）： 805 - 820 .

KIM S ， KWON SH ， KAM TI ， et al . Transneuronal propagation of pathologic α-Synuclein from the gut to the brain models Parkinson's disease ［J］. Neuron ， 2019 ， 103 （ 4 ）： 627 - 641 .

ERNY D ， HRABĚ DE ANGELIS AL ， JAITIN D ， et al . Host microbiota constantly control maturation and function of microglia in the CNS ［J］. Nat Neurosci ， 2015 ， 18 （ 7 ）： 965 - 977 .

SCHEPERJANS F ， AHO V ， PEREIRA PA ， et al . Gut microbiota are related to Parkinson's disease and clinical phenotype ［J］. Mov Disord ， 2015 ， 30 （ 3 ）： 350 - 358 .

XU W ， ZHENG D ， LIU Y ， et al . Glaucocalyxin B alleviates lipopolysaccharide-induced Parkinson's disease by inhibiting TLR/NF-κB and activating Nrf2/HO-1 pathway ［J］. Cell Physiol Biochem ， 2017 ， 44 （ 6 ）： 2091 - 2104 .

YANG J ， JIA M ， ZHANG X ， et al . Calycosin attenuates MPTP-induced Parkinson's disease by suppressing the activation of TLR/NF-κB and MAPK pathways ［J］. Phytother Res ， 2019 ， 33 （ 2 ）： 309 - 318 .

杨明会，李军艳，刘毅 . 系统论引领帕金森病治疗思路［J］. 医学与哲学（临床决策论坛版）， 2009 ， 30 （ 2 ）： 3 - 5 .

李明伟，杨文学，齐小荣，等 . 补肾活血颗粒治疗帕金森病非运动症状的临床研究［J］. 中华保健医学杂志， 2023 ， 25 （ 1 ）： 24 - 27 .

KOLACHEVA A ， BANNIKOVA A ， PAVLOVA E ， et al . Modeling of the Progressive degradation of the nigrostriatal dopaminergic system in mice to study the mechanisms of neurodegeneration and neuroplasticity in Parkinson's disease ［J］. Int J Mol Sci ， 2022 ， 24 （ 1 ）： 683 .

TRAVAGLI RA ， BROWNING KN ， CAMILLERI M . Parkinson disease and the gut： new insights into pathogenesis and clinical relevance ［J］. Nat Rev Gastroenterol Hepatol ， 2020 ， 17 （ 11 ）： 673 - 685 .

栾振先，谌盈帆，秦丽，等 . 基于细胞自噬研究补肾活血方抑制帕金森病模型小鼠α-突触核蛋白集聚的作用［J］. 中国中西医结合杂志， 2020 ， 40 （ 5 ）： 602 - 607 .

李晨，王鹏，王亮，等 . 补肾活血颗粒对亚急性帕金森病模型小鼠脑黑质多巴胺神经元铁死亡的影响［J］. 中医杂志， 2022 ， 63 （ 15 ）： 1463 - 1469 .

陈齐 . 中药药理研究方法学［M］. 北京：人民卫生出版社， 2011 ： 7 .

HARSANYIOVA J ， BUDAY T ， KRALOVA TRANCIKOVA A . Parkinson′s disease and the gut： future perspectives for early diagnosis ［J］. Front Neurosci ， 2020 ， 14 ： 626 . doi： 10.3389/fnins.2020.00626 http://dx.doi.org/10.3389/fnins.2020.00626 .

ELFIL M ， KAMEL S ， KANDIL M ， et al . Implications of the gut microbiome in Parkinson′s disease ［J］. Mov Disord ， 2020 ， 35 （ 6 ）： 921 - 933 .

QIAN YW ， YANG XD ， XU SQ ， et al . Alteration of the fecal microbiota in Chinese patients with Parkinson′s disease ［J］. Brain Behav Immun ， 2018 ， 70 ： 194 - 202 . doi： 10.1016/j.bbi.2018.02.016 http://dx.doi.org/10.1016/j.bbi.2018.02.016 .

SUN MF ， SHEN YQ . Dysbiosis of gut microbiota and microbial metabolites in Parkinson's disease ［J］. Ageing Res Rev ， 2018 ， 45 ： 53 - 61 . doi： 10.1016/j.arr. 2018. 04.004 http://dx.doi.org/10.1016/j.arr.2018.04.004 .

SAMPSON TR ， CHALLIS C ， JAIN N ， et al . A gut bacterial amyloid promotes α-synuclein aggregation and motor impairment in mice ［J］. Elife ， 2020 ， 9 ： e53111 . doi： 10.7554/eLife.53111 http://dx.doi.org/10.7554/eLife.53111 .

NIELSEN HH ， QIU J ， FRIIS S ， et al . Treatment for Helicobacter pylori infection and risk of Parkinson's disease in Denmark ［J］. Eur J Neurol ， 2012 ， 19 （ 6 ）： 864 - 869 .

SAMPSON TR ， DEBELIUS JW ， THRON T ， et al . Gut microbiota regulate motor deficits and neuroinflammation in a model of Parkinson's disease ［J］. Cell ， 2016 ， 167 （ 6 ）： 1469 - 1480 .

MANSUY-AUBERT V ， RAVUSSIN Y . Short chain fatty acids： the messengers from down below ［J］. Front Neurosci ， 2023 ， 17 ： 1197759 . doi： 10.3389/fnins. 2023. 1197759 http://dx.doi.org/10.3389/fnins.2023.1197759 .

CHENG H ， ZHANG D ， WU J ， et al . Atractylodes macrocephala Koidz. volatile oil relieves acute ulcerative colitis via regulating gut microbiota and gut microbiota metabolism ［J］. Front Immunol ， 2023 ， 14 ： 1127785 . doi： 10.3389/fimmu.2023.1127785 http://dx.doi.org/10.3389/fimmu.2023.1127785 .

ZHANG P ， LIU J ， XIONG B ， et al . Microbiota from alginate oligosaccharide-dosed mice successfully mitigated small intestinal mucositis ［J］. Microbiome ， 2020 ， 8 （ 1 ）： 112 .

CAO C ， WANG L ， AI C ， et al . Impact of Lycium barbarum arabinogalactan on the fecal metabolome in a DSS-induced chronic colitis mouse model ［J］. Food Funct ， 2022 ， 13 （ 16 ）： 8703 - 8716 .

JANG YJ ， KIM WK ， HAN DH ， et al . Lactobacillus fermentum species ameliorate dextran sulfate sodium-induced colitis by regulating the immune response and altering gut microbiota ［J］. Gut Microbes ， 2019 ， 10 （ 6 ）： 696 - 711 .

LAGKOUVARDOS I ， LESKER TR ， HITCH T ， et al . Sequence and cultivation study of Muribaculaceae reveals novel species， host preference， and functional potential of this yet undescribed family ［J］. Microbiome ， 2019 ， 7 （ 1 ）： 28 .

查看原文

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effects of Xiaochengqi Decoction （小承气汤） on Intestinal Flora in Rats with Acute Severe Traumatic Brain Injury

Professor ZHOU Shaohua's Experience in Staged Treatment of Parkinson's Disease Based on the "Disease-Symptom-Syndrome Integration" Approach

Treatment of Acute Respiratory Distress Syndrome from the Perspective of Qi Deficiency and Blood Stasis

Clinical Research and Basic Study on Effect of Huangqin Qingre Chubi Capsule （黄芩清热除痹胶囊） on Self-Perception of Patients and Immune Inflammation in Rheumatoid Arthritis

Ganye Pingchan Decoction （感夜平颤方） for Patients with Early-Moderate Parkinson's Disease Complicated with Rapid Eye Movement Sleep Behavior Disorder on Sleep and Blood Pressure Circadian Rhythm Disorders： A Randomized Controlled Trial

Related Author

Lingling PENG

Renjiu LIANG

Ling NING

Yiheng DU

Guihua HUANG

XU Dinglin

WANG Jiayi

GE Liwei

Related Institution

Guangxi University of Chinese Medicine

The First Affiliated Hospital of Guangxi Medical University

Hunan University of Chinese Medicine

Xiyuan Hospital，China Academy of Chinese Medical Sciences

Graduate School，Beijing University of 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

Effects of Bushen Huoxue Formula （补肾活血方） on TLR/NF-κB Pathway and Intestinal Flora in Ileum Tissue of Parkinson's Disease Model Mice

DOI：10.13288/j.11-2166/r.2024.10.011

摘要

Abstract

关键词

Keywords

references