RESUMEN
The study aims to investigate the mechanism of Fengshining fecal microbiota transplants in the intervention of rheumatoid arthritis by ultra-performance liquid chromatography-quadrupole/electrostatic field obitrap high-resolution mass spectrometry (UHPLC-Q-Exactive Orbitrap-MS). All animal welfare and experimental procedures followed the regulations of the Medical Ethics Committee of Shanxi University of Chinese medicine. The rats were randomly divided into normal group, model group, fecal microbiota transplantation group and Tripterygium wilfordii polyglycoside group, and the collagen induced arthritis (CIA) was established. The changes of body weight and metatarpodal lesions of rats were evaluated. The serum of rats in each group was analyzed by liquid chromatography-mass spectrometry and metagenomic technology for differential metabolites and microflora. The protein expression levels of Toll-like receptors (TLR4), myeloid differentiation factor 88 (MyD88) and nuclear factor of kappa B (NF-κB p65) were detected by Western blot. A total of 13 different metabolites, including arachidonic acid, docosahexaenoic acid, 13S-hydroxyoctadecanodienoic acid and L-phenylalanine were screened from serum. Three metabolic pathways, including phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and arachidonic acid metabolism were identified through pathway enrichment. Metagenomic analysis showed that the abundance of g_Bacteroides, g_Prevotella and p_Actinobacteria in CIA group was higher. The abundance of c_Clostridia, g_Akkermansia and s_Akkermansia_muciniphila in fecal microbiota transplantation group is higher. The hierarchical cluster heat map showed that Akkermansia was negatively correlated with L-phenylalanine; while positively correlated with docosahexaenoic acid. Prevotella was positively correlated with L-phenylalanine. Fecal microbiota transplantation group could significantly inhibit the expression of TLR4, MyD88 and p65 proteins in the synovium of rats (P < 0.01). The anti-rheumatoid arthritis effects of fecal microbiota transplantation group is closely related to the intervention of the metabolism of phenylalanine and arachidonic acid, through Akkermansia, Prevotella and other microorganisms, inhibition the TLR4/MyD88/NF-κB pathway.
RESUMEN
This study was aimed to observe the effects ofFeng-Shi-Ning (FSN) capsule on the balance of IL-17+CD4+T (Th17) / Foxp3+CD4+CD25+regulatory T (Treg) cells in peripheral blood and the inflammatory media in synovial fluid of rats with rheumatoid arthritis (RA), in order to further study the mechanism of FSN capsule in RA treatment. The collagen-induced arthritisⅡ (CIA) was used in the RA rat model establishment. The balance of Th17 and Treg cells in peripheral blood was analyzed by the flow cytometry method for effects of FSN capsule treatment. The levels of TNF-α, IL-6, IL-17A, IL-4 and IL-10 in synovial fluid were measured by cytometric bead array (CBA) method. The effects of the normal rats, CIA model rats and the rats treated by tripterygium wilfordii polyglycoside tablet (TPT) were compared. The results showed that compared with the model group, the percentage of Th17 cells in both the FSN capsule group and high-dose CIA group were obviously decreased (P 0.05). There was no significant difference on Th17 and Treg cells among each treatment group and TPT group (P > 0.05). Compared with the model group, the high-dose and middle dose FSN can obviously reduce the levels of TNF-α, IL-6 and IL-17A in synovial fluid of rats in each FSN group (P 0.05). Compared with the TPT group, there were no significant differences on IL-6, TNF-α, IL-17A, IL-4 and IL-10 in each treatment group (P > 0.05). It was concluded that FSN can obviously reduce the percentage of Th17, decrease the secretion of IL-6, TNF-α and IL-17A in synovial fluid. However, it had no obvious effects on the percentage of Treg cells, or the secretion of IL-4 and IL-10 in synovial fluid. The mechanism of FSN capsule in RA treatment may be through regulating Th17 cells, adjusting body immune imbalance, and inhibiting the excessive secretion of inflammatory media in synovial fluid.
RESUMEN
10 and Foxp3 expression, reduce the IL-17 and ROR-γt expression and adjust Treg/Th17 immune balance; effect of 0.1 mg/mL FSN +0.1 mg/mL MTX is the most obvious.