口服硫酸链霉素对帕金森小鼠症状的改善及其对肠道菌群的影响

Effect of oral administration of streptomycin sulfate on symptoms and gut microbiota of Parkinson’s disease mice

【目的】探讨硫酸链霉素对慢性帕金森病(Parkinson’s disease,PD)小鼠症状的改善及肠道菌群的影响。【方法】将40只C57BL/6小鼠分为正常对照组、硫酸链霉素对照组、帕金森病模型组和硫酸链霉素干预帕金森病模型组。帕金森病模型组在实验的前5周采用1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,25 mg/kg)和丙磺舒(250 mg/kg)联合诱导帕金森病模型;硫酸链霉素干预帕金森病模型组在模型构建的同时连续饮用链霉素水溶液(500μg/mL)至实验第8周末。综合运用转棒实验、爬杆实验、免疫组织化学、荧光定量PCR、高通量测序等多种实验方法,检测各组实验小鼠相关症状与指标。【结果】慢性PD模型小鼠与正常对照小鼠相比,表现出极显著的运动障碍(P<0.01),脑黑质纹状体中多巴胺(Dopamine,DA)能神经元及其纤维极显著减少(P<0.01),肠道出现显著的功能紊乱和炎症,同时肠道菌群结构发生了显著的变化:厚壁菌门/拟杆菌门(Firmicutes/Bacteroidetes,F/B)比值升高,疣微菌科(Ruminococcaceae)丰度极显著增加(P<0.01),普雷沃氏菌科(Prevotellaceae)及Prevotellaceae_UCG-001丰度极显著降低(P<0.01)。硫酸链霉素干预显著提高慢性PD小鼠的运动能力(P<0.05),缓解脑黑质纹状体系统中DA能神经元及其纤维的减少(P<0.05),改善肠道功能障碍和肠道炎症,同时可降低F/B的比值,显著降低Ruminococcaceae、理研菌科(Rikenellaceae)和乳杆菌科(Lactobacillaceae)的丰度,增加Prevotellaceae和Prevotellaceae_UCG-001的丰度。【结论】硫酸链霉素可改善PD小鼠相关症状,且影响了PD小鼠的肠道菌群结构。

[Objective]To investigate the effect of streptomycin sulfate on the symptoms and gut microbiota of chronic Parkinson’s disease mice.[Methods]Forty C57BL/6 mice were randomly divided into the control group,streptomycin sulfate control group,Parkinson’s disease(PD)model group and streptomycin sulfate treated PD model group.PD mice were induced by injecting 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(25 mg/kg)combined with probenecid(250 mg/kg)at the first 5 weeks of the experiment;the streptomycin sulfate treated PD mice began to drink the streptomycin aqueous solution(500μg/mL)on the first day of the experiment until the end of the experiment.A variety of experimental methods,such as rotarod test,pole test,immunohistochemistry,real-time PCR,and high-throughput sequencing were used to detect the related symptoms and indicators of each group.[Results]Compared with the control,chronic PD mice showed motor impairment(P<0.01),dopaminergic neurons in the nigrostria and their fibers in the striatum reduced significantly(P<0.01),intestinal dysfunction and inflammation,along with the structure of gut microbiota changed significantly:the ratio of Firmicutes/Bacteroides and the abundance of Family Ruminococcaceae increased(P<0.01),whereas the abundance of Family Prevotellaceae and Genus Prevotellaceae_UCG-001 decreased(P<0.01).The intervention of streptomycin sulfate significantly improved the exercise capacity of chronic PD mice(P<0.05),relieved the decrease of dopaminergic neurons and fibers in the nigrostriatal system(P<0.05),and improved the intestinal dysfunction and inflammation,meanwhile reduced the ratio of the Firmicutes/Bacteroides and the abundance of Family Ruminococcaceae,Rikenellaceae and Lactobacillaceae and increased the abundance of Family Prevotellaceae and Genus Prevotellaceae_UCG-001.[Conclusion]Streptomycin sulfate can relieve the symptoms associated with PD mice and affect the gut microbiota of PD mice.