杜仲方改善帕金森小鼠的肠道微环境

目的] 基于肠-脑轴探讨杜仲方(DZF)对帕金森病(PD)小鼠症状的改善及对其肠道微环境的影响。方法] 40只雄性C57BL/6小鼠随机分为对照组(Control)、模型组(Model)、杜仲方低剂量组(DZF-10 g/kg)、杜仲方高剂量组(DZF-20 g/kg),每组10只。除对照组外,其余各组每3.5 d腹腔注射1次1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP,25 mg/kg),1 h后皮下注射1次丙磺舒(250 mg/kg),共注射10次,对照组注射等量生理盐水。造模开始后,治疗组每天灌胃给予杜仲方,持续至末次造模后1周。分别于第1、3、6周进行爬杆、转棒实验;实验结束后,处死小鼠并取脑,检测小鼠中脑多巴胺能神经元数量及酪氨酸羟化酶(TH)表达水平;取结肠组织,观察结肠组织病理及Occludin、MPO、CD13表达情况;无菌条件下取粪便,16sRNA测序检测小鼠肠道菌群结构变化。结果] 杜仲方干预可明显改善PD小鼠的运动功能,缓解中脑黑质多巴胺能神经元的减少,改善肠道屏障功能和肠道炎症。同时能调整肠道菌群多样性,升高拟杆菌门(Bacteroidetes)丰度,降低厚壁菌门(Firmicutes)丰度,降低厚壁菌门/拟杆菌门(F/B)比值。结论] 杜仲方可以改善帕金森病小鼠的运动障碍及保护多巴胺能神经元,并且可以通过维持肠屏障功能,降低肠道炎症反应,改善肠道菌群结构影响肠道微环境。

Duzhong Formula improves the intestinal microenvironment of Parkinson's mice

Objective] To investigate the effect of Duzhong Formula(DZF) on the improvement of symptoms and intestinal microenvironment of Parkinson''s disease(PD) mice.Methods] Forty male C57BL/6 mice were randomly divided into Control group,Model group,low dose group (DZF-10 g/kg) and high dose group (DZF-20 g/kg),with 10 mice in each group. Except for the Control group,the other groups were intraperitoneally injected with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP,25 mg/kg) every 3.5 days,and subcutaneously injected with probenecid (250 mg/kg) 1 hour later for 10 times. The Control group was injected with the same volume of normal saline. From the beginning of modeling,the treatment group was given DZF by gavage every day until one week after the last modeling. The pole climbing and rod rotating were performed at 1,3,and 6 weeks. At the end of the experiment,the mice were sacrificed and the brains were removed for determination of the number of dopaminergic neurons in the midbrain and the expression of tyrosine hydroxylase (TH). The colonic tissues were collected for pathological examination of Occludin,MPO,and CD13. Fecal samples were collected,and 16sRNA sequencing was used to detect the structural changes of gut microbiota in mice.Results] DZF could significantly improve the motor function of PD mice,alleviate the decrease of dopaminergic neurons,improve the intestinal barrier function and intestinal inflammation. At the same time,it could adjust the diversity of intestinal flora,increase the abundance of Bacteroidetes(B),reduce the abundance of Firmicutes(F),and reduce the F/B ratio.Conclusion] DZF can improve the dyskinesia and protect dopaminergic neurons in mice with Parkinson''s disease,and can affect the intestinal microenvironment by maintaining intestinal barrier function,reducing intestinal inflammation and improving intestinal microbiota structure.