健脾益肾通络方对MPTP诱导的帕金森病小鼠肠道菌群与短链脂肪酸的调节作用

探讨健脾益肾通络方（JYTR）对1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）诱导的帕金森病小鼠肠道菌群组成和短链脂肪酸的影响。

将成年雄性C57BL/6小鼠随机分成3组：空白组、MPTP组、MPTP+JYTR组，每组15只。采用爬杆、悬挂、横梁实验评估运动情况。采用免疫荧光法检测纹状体酪氨酸羟化酶（TH）的表达。采用蛋白印迹法检测纹状体和结肠中α-突触核蛋白（α-syn）的表达。采用16S核糖体RNA（16S rRNA）基因测序评估肠道菌群的变化。采用气相色谱质谱法（GC-MS）分析小鼠粪便短链脂肪酸的水平。最后对差异菌群和短链脂肪酸进行相关性分析。

与MPTP组相比，MPTP+JYTR组小鼠悬挂评分增加（P=0.040），爬杆时间和横梁行走时间显著缩短（P=0.042、P＜0.001）；纹状体TH表达显著增加（P=0.045），纹状体、结肠中α-syn表达显著减少（P=0.011、P=0.037）；肠道菌群alpha多样性、beta多样性显著改变；门水平方面拟杆菌门丰度增加（P=0.003），厚壁菌门、疣微菌门丰度均显著减少（P=0.001、P=0.011）；科水平方面毛螺菌科丰度显著升高（P=0.001），嗜黏蛋白阿克曼菌科、丹毒丝菌科丰度均显著降低（P=0.008、P=0.004）；短链脂肪酸中乙酸、丙酸水平均显著增加（P=0.014、P=0.010）。相关性分析显示爬杆时间、悬挂评分、横梁时间均与肠道菌群、短链脂肪酸存在相关性。

JYTR可以通过调节肠道菌群失调和改善短链脂肪酸水平起到治疗帕金森病的作用。

Regulation of intestinal flora and short-chain fatty acids in MPTP-induced Parkinson's mice by Jianpi Yishen Tongluo Recipe

ObjectiveTo observe the effects of Jianpi Yishen Tongluo Recipe (JYTR) on the compositions of intestinal flora and short-chain fatty acids (SCFAs) in mice with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease. MethodsC57BL/6 mice were randomly divided into three groups: Control group (n=15), MPTP group (n=15) and MPTP+JYTR group (n=15). The movement was assessed by using pole climbing, suspension and beam experiments. Striatal tyrosine hydroxylase (TH) expression was detected with immunofluorescence. Protein blotting was used to detect α-synuclein (α-syn) expression in striatum and colon. 16S ribosomal RNA gene sequencing was used to assess the changes in the gut microbiota. Gas chromatography-mass spectrometry was used to analyze the content of SCFAs in mouse feces. Finally, correlation analysis was performed on the differential flora and SCFAs. ResultsCompared to the MPTP-treated group, mice in the MPTP+JYTR group exhibited an elevation in tail suspension test scores (P=0.040), alongside a pronounced decrease in both pole-climbing (P=0.042) and beam-walking times (P＜0.001). There was also a significant uptick in striatal TH expression (P=0.045), coupled with a notable dip in α-syn levels in both the striatum and colon (P=0.011, P=0.037). In terms of gut microbiota, there was a noteworthy augmentation in both alpha and beta diversity; phylum-level analyses revealed a surge in Bacteroidota abundance (P=0.003), whereas populations of Firmicutes and Verrucomicrobiota saw a significant downturn (P=0.001, P=0.011). At the family level, there was a significant influx in Lachnospiraceae (P=0.001), while the relative abundance of Akkermansiaceae and Erysipelotrichaceae diminished substantially (P=0.008, P=0.004). Additionally, concentrations of the SCFAs acetate and propionate were significantly elevated (P=0.014, P=0.010). Correlation analyses suggested that the improvements in pole climbing and tail suspension tests, as well as beam walking times, were associated with shifts in the gut microbiota composition and SCFAs profiles. ConclusionJYTR may play a role in the treatment of Parkinson's disease by regulating the dysbiosis of intestinal flora and improving the levels of SCFAs.