Objective To explore the possible mechanism of Bushen Huoxue Formula(补肾活血方,BHF)in the treatment of Parkinson's disease(PD)from the the perspective of intestinal flora.Methods Seventy-two male C57/BL6J mice were randomly divided into blank group,model group,Madopar group and low-,medium-and high-dose BHF groups,with 12 mice in each group.The mice in the blank group were intraperitoneally injected with 10 ml/kg of normal saline,and those in the other groups were intraperitoneally injected with 30 mg/kg of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)at a concentration of 3 mg/ml to induce PD mice model,both once a day for 7 consecu-tive days.After successful modeling,the low-,medium-,and high-dose BHF groups were given 7.5,15,and 30 g/(kg·d)of BHF by gavage,respectively,while the Madopar group was given 112.5 mg/(kg·d)of Domedopar tablets by gavage,and the blank group and the model group were given 15 ml/(kg·d)of distilled water,all once a day for 14 consecutive days.The rod climbing test,rotating rod test,grip strength test and weight-bearing swimming test were used to evaluate the behavioral indicators of mice.Western blotting was used to measure the protein expression levels of Toll-like receptor(TLR)/nuclear factor kappa B(NF-κB)pathway inflammatory factors in the mouse ileum,includ-ing Toll-like receptor 2(TLR2),Toll-like receptor 4(TLR4),NF-κB,tumor necrosis factor α(TNF-α),inter-leukin 6(IL-6),and interleukin 17(IL-17).16S rRNA high-throughput sequencing was used to analyze changes in mouse intestinal flora.Results Compared to those in the blank group,the mice in the model group had longer bot-toming time when climbing the pole,reduced grip strength,shortened rotary pole duration and swimming duration,and increased protein expression levels of TLR2,TLR4,NF-KB,TNF-α,and IL-6 in the ileal tissue(P＜0.01).Compared to the model group,the Madopar group and the low-,medium-and high-dose BHF groups had shortened bottoming time of the climbing pole and increased grip strength;the Madopar group and the high-dose BHF group had prolonged rotary pole duration,and reduced protein expressions of TLR2,TLR4,NF-κB,TNF-α,IL-6,and IL-17 levels;and only the high-dose BHF group had prolonged swimming duration(P＜0.05 or P＜0.01).Compared to those in the low-dose BHF group,the bottoming time of the climbing pole were shorter in the moderate-and high-dose groups(P＜0.05 or P＜0.01),and the grip strength increased while the protein expression levels of TLR2,TLR4 and IL-17 decreased in the high-dose group(P＜0.05 or P＜0.01).The intestinal flora results showed significant differences between the blank group and the model group in the Dominance index,Pielou_e index,Shannon index,and Simpson index(P＜0.05 or P＜0.01).Compared to those of the model group,the Shannon index,Chao1 index,and Observed_otus index of the Madopar group,as well as the Chao1 index,Observed_otus index,Dominance index,Pielou_e index,Shannon index,and Simpson index of the high-dose BHF group all showed significantly statistical differences(P＜0.05 or P＜0.01).At the phylum level,the relative abundance categories of bacterial phyla with statistically significant differences in each group included Proteobacteria,Bacteroidetes,and Firmicutes(P＜0.05 or P＜0.01).At the genus level,the relative abundance categories of bacterial genera with statistically significant diffe-rences among each group included Muribaculaceae,Akkermansia,and Helicobacter pylori(P＜0.05 or P＜0.01).Conclusion The possible mechanism of BHF in treating PD may be to reconstruct the disordered intestinal flora structure and improve the inflammatory response.
Parkinson's diseaseBushen Huoxue Formula(补肾活血方)intestinal florainflammatory responseToll-like receptorsnuclear factor kappa B
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