Liriodendrin exerts protective effects against chronic endometritis in rats by modulating gut microbiota composition and the arginine/nitric oxide metabolic pathway.

BACKGROUND: Chronic endometritis (CE), a gynecological disease, is characterized by inflammation. Liriodendrin is reported to exhibit anti-inflammatory properties. However, the therapeutic effects of liriodendrin on CE and the underlying molecular mechanisms have not been elucidated. This study aimed to investigate the therapeutic effects of liriodendrin on CE in rats and the underlying mechanisms. METHODS: A CE rat model was established and administered with liriodendrin for 21 days. The serum levels of inflammatory cytokines were examined using enzyme-linked immunosorbent assay. The uterine mRNA levels of cytokines were examined using quantitative real-time polymerase chain reaction analysis. The activation of the Toll-like receptor 4 (TLR4)/NF-κB pathway was investigated using western blotting analysis. The effects of liriodendrin on intestinal flora and serum metabolites were examined using 16S rRNA sequencing and untargeted serum metabolomics, respectively. The protein and mRNA levels of arginase-2 (Arg-2) and the nitric oxide (NO) metabolic pathway-related factors were assessed. Molecular docking was performed to explore the interaction between liriodendrin and Arg-2. RESULTS: Liriodendrin alleviated the CE-induced pathological changes in the uterus, modulated the serum levels of inflammatory cytokines, and downregulated the mRNA and protein levels of TLR4/NF-κB pathway-related factors. Treatment with liriodendrin mitigated the CE-induced upregulation of Firmicutes/Bacteroidetes ratio and Lachnospiraceae abundance and downregulation of Ruminococcaceae abundance. Serum metabolomic analysis revealed that liriodendrin regulated the biosynthesis of choline metabolism pathway-related factors. Liriodendrin suppressed the CE-induced upregulation of Arg-2 and downregulation of inducible nitric oxide synthase (iNOS) expression, and NO levels by directly binding to the amino acid residues of Arg-2 through hydroxyl bonds. CONCLUSIONS: Liriodendrin exerted therapeutic effects on CE in rats through the alleviation of inflammation by modulating the gut microbiota structure, directly downregulating Arg-2, and regulating the arginine/NO metabolic pathway.

Effect of bushenyisui formula on brain tissue apoptosis and Bcl-2 in beta-amyloid protein-induced Alzheimer's disease rat models.

OBJECTIVE: To investigate the effect of Bushenyisui Formula on cell apoptosis and positive B cell lymphoma (Bcl-2) in the Brain of rat models of Alzheimer's disease (AD) induced by beta-amyloid protein (Abeta) and the mechanism underlying the effect. METHODS: Total of 40 SD rats, 20 females and 20 males, were randomly assigned to 4 groups, controlled group (A), model group (B), conventional treatment group (C) and Bushenyisui Formula treatment (BYFT) group (D), 10 rats in each group. Abeta 1-42 was injected into the bilateral hippocampus of the rats in group B, C and D to create the models of AD. Sham operation was performed on the rats of group A in the same way by injecting equal volume of 0.9% sodium chloride solution into their bilateral hippocampus. 5 days after operation, Bushenyisui Formula was intraperitoneally administered at a dose of 450 mg/kg to the rats of group D (QD) for 20 days. Equal volume of 0.9% sodium chloride solution was intraperitoneally injected into the rats of group B with the same procedure. C suspension (20 mg/mL) was intraperitoneally injected into the rats of group B with the same procedure. The number of apoptotic cells in Brain and the positive Bcl-2 were counted. The changes of learning and memory abilities were evaluated using Y-maze. RESULTS: Right after the establishment of the models, group B, C and D compared to group A respectively, the outcomes of Y-maze were significantly different from that of group A, which suggested obvious learning and memory disorder in those groups (P < 0.01). After treatment, the times of electronic shocks of group C and D were significantly less than that of group B (P < 0.05), and the numbers of apoptotic cells and positive Bcl-2 were significantly different from those of group B, apoptotic sells' number of group C and D smaller than that of group B and the number of positive Bcl-2 greater than that of group B. CONCLUSION: Bushenyisui Formula could increase the number of Bcl-2 in brain, which improved the function of nervous system pertaining to learning and memory abilities.

Effects of tongmai huoxue yin (see text) on tumor necrosis factor-alpha in the acute cerebral ischemia model rat.

OBJECTIVE: To observe the interfering action of Tongmai Huoxue Yin (see text) on the acute cerebral ischemia model rat. METHODS: Total 60 SD rats, 30 females and 30 males, were randomly divided into 4 groups, sham-operation group, model group, Nimodipine group and Tongmai Huoxue Yin group, 15 rats in each group. The acute cerebral ischemia rat model was duplicated, the middle cerebral artery (MCA) were ligated and the thread was inserted for the rats in the model group, Nimodipine group and Tongmai Huoxue Yin group, for the rats in the sham-operation group, the arteries were separated without ligature and the thread was not inserted. After the modeling has succeed, the water-decocted concentrated solution of 20-fold Tongmai Huoxue Yin clinical dosage was intragastrically administrated in a dose of 3 mL/100 g d divided into twice, 1.5 mL/100 g once. Distilled water 3 mL/100 g x d was intragastrically administrated, 1.5 mL/100 g once, for the rat in the model group, Nimodipne suspension 3 mL/100 g x d (0.6 mg / 100 g) for the Nimodipine group and 3 mL/100 g x d (5.4 g/100 g) for the Tongmai Huoxue Yin group, no drugs for the sham-operation group. And changes of tumor necrosis factor-alpha (TNF-alpha) contents in the serum and brain tissue were investigated. RESULTS: Compared with the model group, compared with the sham-operation group, serum TNF-alpha content at 5 h of focal cerebral ischemic ischemia in the model group started to increase and reached to the high peak at 12 h, but in both the Tongmai Huoxue Yin group and the Nimodipine group decreased in varying degrees at the same time; compared with the sham-operation group, brain TNF-alpha content at 6 h of focal cerebral ischemic ischemia in the model group started to increase and reached to the high peak at 12 h, but in both the Tongmai Huoxue Yin group and the Nimodipine group decreased in varying degrees, with the most obviously decreased at 24 h of ischemia.Tongmai Huoxue Yin could significantly decrease TNF-alpha content in the brain tissue. CONCLUSION: Tongmai Huoxue Yin has a protective action on acute cerebral ischemia injury in the rat.