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[甘松对帕金森病大鼠肠道微生物群的影响]

[Effects of Nardostachys jatamansi on gut microbiota of rats with Parkinson's disease].

作者信息

Wan Guo-Hui, Wei Xiao-Jia, Li Jia-Yuan, Yang Xue, Yu Jia-He, Liu Jin-Feng, Wang Yu-Qing, Lyu Yan, Jin Zhong-Xian, Shi Jin-Li

机构信息

School of Chinese Materia Medica, Beijing University of Chinese Medicine Beijing 102488, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2022 Jan;47(2):499-510. doi: 10.19540/j.cnki.cjcmm.20210922.701.

Abstract

Under the guidance of the traditional Chinese medicine(TCM) theory of "Zangfu-organs of spleen and stomach" and the modern theory of "microbiota-gut-brain axis", this study explored the effects of Nardostachys jatamansi on the gut microbiota of rats with Parkinson's disease(PD). The 40 SD rats were randomly divided into the control group, PD model group, levodopa group, and Nardostachys jatamansi ethanol extract group. The PD model was established by subcutaneous injection of rotenone in the neck and back area. After 14 days of intragastric administration, the PD rats' behaviors were analyzed through open field test, inclined plane test, and pole test. After the behavioral tests, the striatum, colon, and colon contents of rats in each group were collected. Western blot was employed to detect the protein expression of tyrosine hydroxylase(TH) and α-synuclein(α-syn) in striatum and that of α-syn in colon. Enzyme linked immunosorbent assay(ELISA) was used to detect the levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and nuclear factor-kappa B(NF-κB) in striatum and colon. High-throughput sequencing of 16 S rRNA gene was conducted to detect the differences in microbial diversity, abundance, differential phyla, and dominant bacteria of rats between groups. The results indicated that Nar. ethanol extract could relieve dyskinesia, reverse the increased levels of α-syn, TNF-α, IL-1β, and NF-κB in striatum, and improve the protein expression of TH in striatum of PD rats. The α diversity analysis indicated a significant decrease in diversity and abundance of gut microbiota in the PD model. The results of linear discriminant analysis effect size(LEfSe) of dominant bacteria indicated that Nardostachys jatamansi ethanol extract increased the relative abundance of Clotridiaceae, Lachnospiraceae, and Anaerostipes, and reversed the increased relative abundance of Proteobacteria, Gammaproteobacteria, Enterobacteriaceae, and Escherichia-Shigella in PD model group to exhibit the neuroprotective effect. In summary, the results indicated that Nar. ethanol extract exert the therapeutic effect on PD rats. Specifically, the extract may regulate gut microbiota, decrease the levels of proinflammatory cytokines, and reduce the protein aggregation of α-syn in the colon and striatum to alleviate intestinal inflammation and neuroinflammation. This study provides a basis for combining the theory of "Zangfu-organs of spleen and stomach" with the theory of "microbiota-gut-brain axis" to treat PD.

摘要

本研究在中医“脾胃脏腑”理论和现代“微生物群-肠道-脑轴”理论的指导下,探讨了甘松对帕金森病(PD)大鼠肠道微生物群的影响。将40只SD大鼠随机分为对照组、PD模型组、左旋多巴组和甘松乙醇提取物组。通过在颈部和背部皮下注射鱼藤酮建立PD模型。灌胃给药14天后,通过旷场试验、斜面试验和杆试验分析PD大鼠的行为。行为测试后,收集各组大鼠的纹状体、结肠及结肠内容物。采用蛋白质免疫印迹法检测纹状体中酪氨酸羟化酶(TH)和α-突触核蛋白(α-syn)以及结肠中α-syn的蛋白表达。采用酶联免疫吸附测定(ELISA)法检测纹状体和结肠中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和核因子-κB(NF-κB)的水平。对16S rRNA基因进行高通量测序,以检测各组大鼠微生物多样性、丰度、差异菌门和优势菌的差异。结果表明,甘松乙醇提取物可缓解运动障碍,逆转纹状体中α-syn、TNF-α、IL-1β和NF-κB水平的升高,并改善PD大鼠纹状体中TH的蛋白表达。α多样性分析表明,PD模型中肠道微生物群的多样性和丰度显著降低。优势菌的线性判别分析效应大小(LEfSe)结果表明,甘松乙醇提取物增加了梭菌科、毛螺菌科和厌氧短杆菌属的相对丰度,并逆转了PD模型组中变形菌门、γ-变形菌纲、肠杆菌科和埃希氏菌-志贺氏菌属相对丰度的增加,从而发挥神经保护作用。总之,结果表明甘松乙醇提取物对PD大鼠具有治疗作用。具体而言,该提取物可能调节肠道微生物群,降低促炎细胞因子水平,并减少结肠和纹状体中α-syn的蛋白聚集,以减轻肠道炎症和神经炎症。本研究为将“脾胃脏腑”理论与“微生物群-肠道-脑轴”理论相结合治疗PD提供了依据。

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