异功散通过调控肠道微生物改善痴呆症模型大鼠的认知下降

doi:10.12122/j.issn.1673-4254.2024.07.09

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (7): 1297-1305.doi: 10.12122/j.issn.1673-4254.2024.07.09

• • 上一篇

异功散通过调控肠道微生物改善痴呆症模型大鼠的认知下降

曾静¹^,²(), 陈荣¹^,², 任香怡³, 花雷¹^,², 阳勇¹^,²^,⁵, 魏江平¹^,²^,⁴(), 张小梅¹^,²^,⁵()

^1.中药新药创制川渝共建重点实验室//重庆市中药研究院，重庆 400065
^2.国家中医药管理局中药化学三级实验室//重庆市中药研究院，重庆 400065
^3.湖南中医药大学第一附属医院，长沙 410007
^4.重庆医科大学药学院，重庆 400016
^5.川渝共建慢病中药新药泸州市重点实验室//西南医科大学附属中医院，四川泸州 646000

收稿日期:2024-04-11 出版日期:2024-07-20 发布日期:2024-07-25
通讯作者: 魏江平,张小梅 E-mail:1589905936@qq.com;sichuanwjp@163.com;ZXM761@163.com
作者简介:曾静，硕士，E-mail: 1589905936@qq.com
基金资助:
重庆市技术创新与应用发展专项重点项目(cstc2021jscx-dxwtBX0012);重庆市自然科学基金项目博士后项目(cstc2021jscx-bshX0054);重庆市中药研究院基本科研业务费项目(jbky20210003);湖南省卫健委一般指导课题项目(D202313058007)

Yigong San improves cognitive decline in a rat model of Alzheimer's disease by regulating intestinal microorganisms

Jing ZENG¹^,²(), Rong CHEN¹^,², Xiangyi REN³, Lei HUA¹^,², Yong YANG¹^,²^,⁵, Jiangping WEI¹^,²^,⁴(), Xiaomei ZHANG¹^,²^,⁵()

^1.Sichuan -Chongqing Joint Key Laboratory of Innovation of New Drugs of Traditional Chinese Medicine
^2.Third Level Laboratory of Traditional Chinese Medicine Chemistry of the State Administration of Traditional Chinese Medicine, Chongqing Academy of Chinese Materia Medica, Chongqing 400065, China
^3.First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410007, China
^4.College of Pharmacy, Chongqing Medical University, Chongqing 400016, China
^5.Luzhou Key Laboratory of Traditional Chinese Medicine for Chronic Diseases Jointly Built by Sichuan and Chongqing, Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou 646000, China

Received:2024-04-11 Online:2024-07-20 Published:2024-07-25
Contact: Jiangping WEI, Xiaomei ZHANG E-mail:1589905936@qq.com;sichuanwjp@163.com;ZXM761@163.com

摘要/Abstract

摘要：

目的研究异功散对脂多糖（LPS）诱导认知下降大鼠学习记忆能力的影响，并通过肠道微生物分析其作用机制。方法将40只SD大鼠随机分成空白组、模型组、盐酸多奈哌齐组（1.17 mg/kg）和异功散高剂量组（5.25 g/kg）、异功散低剂量组（2.63 g/kg），8只/组。各组灌胃给予相应药物或纯水干预24 d后，除空白组腹腔注射生理盐水外，其余各组每天腹腔注射LPS（0.5 mg/kg）造模，并开始水迷宫评价其学习记忆能力改变；HE染色观察海马病理形态学变化；16S rRNA检测各组肠道微生物物种变化；酶联免疫吸附测定法检测脑组织和血清中IL-6、TNF-α和IL-1β水平。结果与对照组比较，模型组大鼠第5天逃避潜伏期明显延长（P<0.01），穿越平台次数、平台象限活动累积时间和平均游泳速度均下降，其中穿越平台次数具有统计学差异（P<0.05）；海马区可见神经细胞排列散乱、神经细胞变性或坏死等病变且细胞损伤病理评分明显升高（P<0.01）；脑组织和血清IL-6、TNF-α和IL-1β水平均明显升高（P<0.01）；Alph多样性评价指标如ACE、Chao1等均明显升高（P<0.05），在科水平的Prevotellaceae等物种丰度明显降低而Desulfovibrionaceae等物种丰度则升高。与模型组比较，异功散组第5天的逃避潜伏期均明显缩短（P<0.01），高剂量组穿越平台次数具有统计学差异（P<0.05）；海马区神经细胞变性或坏死等病变均有好转且细胞损伤病理评分明显降低（P<0.05）；脑组织和血清IL-6、TNF-α和IL-1β水平也均明显下降（P<0.01）；Alph多样性评价指标如ACE、Chao1等均明显降低（P<0.05），在科水平的Prevotellaceae等物种丰度明显升高而Desulfovibrionaceae等物种丰度则降低。结论异功散改善认知下降大鼠的学习记忆能力和海马病理形态学等病变可能与调控肠道Prevotellaceae等的丰度，以及信号转导、辅因子和维生素的代谢等代谢途径有关。

关键词: 异功散, 脂多糖, 认知下降, 肠道微生物

Abstract:

Objective To investigate the effect of Yigong San (YGS) on learning and memory abilities of rats with lipopolysaccharide (LPS)‑induced cognitive decline and explore its possible mechanism in light of intestinal microbiota. Methods Forty SD rats were randomly divided into control group, model group, donepezil (1.3 mg/kg) group, and high-dose (5.25 g/kg) and low-dose (2.63 g/kg) YGS treatment groups. After 24 days of treatment with the corresponding drugs or water by gavage, the rats in the latter 4 groups received an intraperitoneal injection of LPS (0.5 mg/kg) to establish models of Alzheimer's disease (AD). Water maze test and HE staining were used to evaluate the changes in learning and memory abilities and pathomorphology of the hippocampus. The changes in gut microbial species of the rats were analyzed with 16S rRNA sequencing, and the levels of IL-6, TNF-α, and IL-1β in the brain tissue and serum were detected using ELISA. Results Compared with the AD model group, the YGS-treated rats showed significantly shortened escape latency on day 5 after modeling, reduced neuronal degeneration and necrosis in the hippocampus, lowered pathological score of cell damage, and decreased levels IL-6, TNF-α and IL-1β in the brain tissue and serum. The YGS-treated rats showed also obvious reduction of Alpha diversity indicators (ACE and Chao1) of intestinal microbiota with significantly increased abundance of Prevotellaceae species at the family level and decreased abundance of Desulfovibrionaceae, which were involved in such metabolic signaling pathways as cell community prokaryotes, membrane transport, and energy metabolism. Conclusion YGS improves learning and memory abilities and hippocampal pathomorphology in AD rat models possibly by regulating the abundance of intestinal microbial species such as Prevotellaceae to affect the metabolic pathways for signal transduction, cofactors, and vitamin metabolism.

Key words: Yigong San, lipopolysaccharide, cognitive decline, intestinal microbiota

曾静, 陈荣, 任香怡, 花雷, 阳勇, 魏江平, 张小梅. 异功散通过调控肠道微生物改善痴呆症模型大鼠的认知下降[J]. 南方医科大学学报, 2024, 44(7): 1297-1305.

Jing ZENG, Rong CHEN, Xiangyi REN, Lei HUA, Yong YANG, Jiangping WEI, Xiaomei ZHANG. Yigong San improves cognitive decline in a rat model of Alzheimer's disease by regulating intestinal microorganisms[J]. Journal of Southern Medical University, 2024, 44(7): 1297-1305.

图/表 6

图1 异功散对LPS诱导大鼠学习记忆能力的影响

Fig.1 Effect of Yigong San on learning and memory ability of rats with LPS-induced Alzheimer's disease (AD). A: Escape latency in hidden platform test (n=8). B: Representative trajectory map of hidden platform test on day 5 after modeling. C: Representative trajectory map of spatial probe test. ^P<0.05, ^^P<0.01, ^^^P<0.001 vs the escape latency on the first day in each group; *P<0.05, **P<0.01, ***P<0.001 vs Control; #P<0.05, ##P<0.01, ###P<0.001 vs Model.

表1 穿越平台次数、平台象限累积时间和平均游泳速度

Tab.1 Number of platform crossings, platform quadrant accumulation time and average swimming speed (Mean±SD)

Group	Dosage	Times of platform crossing time (s)	Platform quadrant cumulative time (s)	Average swimming speed(cm/s)
Control	-	3.85±1.77^##	34.53±10.21^#	14.43±1.77
Model	-	1.43±0.98**	21.49±2.92*	12.71±2.75
Donepezil	1.3 mg/kg	2.00±0.89	24.45±8.71	9.72±2.60
YGS-H	5.25 g/kg	4.14±2.97^#	26.19±6.33	14.32±1.62
YGS-L	2.63 g/kg	3.29±0.95^##	28.81±5.83	14.60±3.27

图2 异功散对LPS诱导痴呆模型大鼠海马病理形态学的影响

Fig.2 Effect of Yigong San on pathomorphology of the hippocampus in rats with LPS-induced AD. A: Control. B: Model. C: YGS-H. D: YGS-L. n=8, *P<0.05 vs Control; ##P<0.01 vs Model.

图3 异功散对LPS诱导痴呆大鼠脑组织和血清TNF-α、IL-1β和IL-6含量变化

Fig.3 Effect of Yigong San on brain tissue and serum levels of TNF-α, IL-1β and IL-6 in rats with LPS-induced AD. n=6, **P<0.01, ***P<0.01 vs Control; #P<0.05, ##P<0.01, ###P<0.001 vs Model.

图4 多样性分析

Fig.4 Diversity analysis of intestinal microbiota of the rats. A: Statistical analysis of alpha diversity index (n=6). B: β-diversity analysis-principal coordinates analysis (PCoA). C: β-diversity analysis-non-metricmulti-dimensional scaling (NMDS). *P<0.05, **P<0.01 vs Control; #P<0.05, ##P<0.01, ###P<0.001 vs Model.

图6 功能基因预测分析

Fig.6 Functional gene prediction analysis. A: KEGG metabolic pathway. B: Analysis of the differences in KEGG metabolic pathways among the groups.

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异功散通过调控肠道微生物改善痴呆症模型大鼠的认知下降

Yigong San improves cognitive decline in a rat model of Alzheimer's disease by regulating intestinal microorganisms

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