The technique of alizarin red staining allowed for the identification of areas of osteoblast mineralization. The model group displayed significantly attenuated cell proliferation and alkaline phosphatase (ALP) activity relative to the control group. This was associated with a decrease in the expression of BK channel subunit (BK), collagen (COL1), bone morphogenetic protein 2 (BMP2), osteoprotegerin (OPG), and phosphorylated Akt, and a reduction in the mRNA levels of Runt-related transcription factor 2 (RUNX2), BMP2, and OPG. Finally, a corresponding decline in the calcium nodule area was observed. Serum containing EXD significantly amplified cellular proliferation and ALP activity, increased protein expression of bone morphogenetic protein 2 (BMP2), collagen 1 (COL1), osteoprotegerin (OPG), phosphorylated Akt, and forkhead box protein O1 (FoxO1), and elevated mRNA levels of runt-related transcription factor 2 (RUNX2), BMP2, and OPG, culminating in an increase in the area of calcium deposits. Reversal of EXD-containing serum's effect on enhancing the protein expression of BK, COL1, BMP2, OPG, and phosphorylated Akt and FoxO1, and boosting mRNA expression of RUNX2, BMP2, and OPG, was observed following BK channel blockage by TEA, along with a resulting expansion of the calcium nodule area. The presence of EXD in serum might improve MC3T3-E1 cell proliferation, osteogenic differentiation, and mineralization capabilities under oxidative stress, likely by affecting BK channel activity and downstream Akt/FoxO1 signaling.
The objective of this study was to ascertain the impact of Banxia Baizhu Tianma Decoction (BBTD) on the cessation of anti-epileptic drugs, and to examine the association between BBTD and alterations in amino acid metabolism through transcriptomic analysis, employing a lithium chloride-pilocarpine-induced epilepsy model in rats. The sample of rats with epilepsy was segmented into a control group (Ctrl), an epilepsy group (Ep), a combined group receiving BBTD and antiepileptic drugs (BADIG), and a group in which antiepileptic drugs were withdrawn (ADWG). For 12 weeks, the Ctrl and Ep groups were given ultrapure water using the gavage technique. Over 12 weeks, the BADIG's treatment included gavage administration of BBTD extract and carbamazepine solution. MZ-101 The ADWG's treatment regimen involved gavage administration of carbamazepine solution and BBTD extract for the first six weeks, and subsequently, only BBTD extract for the subsequent six weeks. The therapeutic effect was determined using a multifaceted approach encompassing behavioral observation, electroencephalogram (EEG) readings, and hippocampal neuronal morphological changes. High-throughput sequencing revealed differential genes linked to amino acid metabolism in the hippocampus, and real-time quantitative polymerase chain reaction (RT-qPCR) validated the mRNA expression levels within the hippocampus for each group. Through protein-protein interaction (PPI) network analysis, the hub genes were identified, followed by Gene Ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Two ceRNA networks, namely circRNA-miRNA-mRNA and lncRNA-miRNA-mRNA, were constructed to compare ADWG and BADIG. The experimental data highlighted a significant improvement in behavioral observation, EEG readings, and hippocampal neuronal integrity in rats of the ADWG group, when compared with the Ep group. RT-qPCR analysis corroborated the transcriptomic findings, which pinpointed thirty-four differential genes involved in amino acid metabolism; the sequencing results were validated. Analysis of the PPI network yielded eight hub genes, each deeply involved in multiple biological processes, molecular functions, and signal transduction pathways, notably those related to amino acid metabolism. A comparison between ADWG and BADIG demonstrated two ternary transcription networks: one featuring 17 circRNAs, 5 miRNAs, and 2 mRNAs, and the other composed of 10 lncRNAs, 5 miRNAs, and 2 mRNAs. In summary, the withdrawal of antiepileptic drugs by BBTD may be attributable to modifications in the transcriptomic regulation of amino acid metabolism.
This research investigated the impact and underlying mechanism of Bovis Calculus in ulcerative colitis (UC), employing a network pharmacology prediction strategy coupled with animal model verification. After utilizing databases such as BATMAN-TCM to pinpoint potential targets of Bovis Calculus against UC, the pathway enrichment analysis was carried out. Seventy healthy C57BL/6J mice were randomly divided into distinct groups based on body weight: a control group, a model group, a 2% polysorbate 80 solvent group, a 0.40 g/kg salazosulfapyridine (SASP) group, and high-, medium-, and low-dose Bovis Calculus Sativus (BCS) groups (0.20, 0.10, and 0.05 g/kg, respectively). Mice were treated with 3% dextran sulfate sodium (DSS) solution daily for a period of seven days to produce the UC model. For three days preceding the modeling procedure, mice assigned to drug intervention groups were administered their corresponding drugs orally (gavage), and this medication continued for seven days during the modeling process (a total of ten days of continuous treatment). The body weight of mice and the disease activity index (DAI) were meticulously tracked and recorded throughout the experiment. The modeling procedure, lasting seven days, was followed by a measurement of the colon's length and the observation of pathological changes within the colon's tissues using hematoxylin-eosin (H&E) staining. To measure the levels of tumor necrosis factor-(TNF-), interleukin-1(IL-1), interleukin-6(IL-6), and interleukin-17(IL-17), an enzyme-linked immunosorbent assay (ELISA) was performed on the colon tissues from the mice. The mRNA expression levels of IL-17, IL-17RA, Act1, TRAF2, TRAF5, TNF-, IL-6, IL-1, CXCL1, CXCL2, and CXCL10 were investigated by using real-time polymerase chain reaction (RT-PCR). antipsychotic medication Western blot analysis was performed to determine the protein expression of IL-17, IL-17RA, Act1, p-p38 MAPK, and p-ERK1/2. Network pharmacological predictions suggest that Bovis Calculus may exert therapeutic effects via the IL-17 and TNF signaling pathways. Animal research on the tenth day of drug treatment demonstrated a considerable rise in body weight, a reduction in DAI score, and an increase in colon length within the BCS treatment groups. This was coupled with an amelioration in colon mucosal damage and a pronounced decrease in TNF-, IL-6, IL-1, and IL-17 expression within colon tissues, when contrasted with the solvent control group. In ulcerative colitis (UC) model mice, high-dose BCS (0.20 g/kg) treatment exhibited a substantial reduction in the mRNA expression of IL-17, Act1, TRAF2, TRAF5, TNF-, IL-6, IL-1, CXCL1, and CXCL2 within colon tissue, a tendency towards decreased mRNA expression of IL-17RA and CXCL10, and a significant inhibition of IL-17RA, Act1, and p-ERK1/2 protein expression. Moreover, the protein expression of IL-17 and p-p38 MAPK also showed a tendency to decrease. Using a whole-organ-tissue-molecular approach, this study, for the first time, demonstrates that BCS might reduce pro-inflammatory cytokine and chemokine expression by inhibiting the IL-17/IL-17RA/Act1 signaling pathway. This treatment improves the inflammatory injury to colon tissues in DSS-induced UC mice, mirroring traditional approaches to clearing heat and removing toxins.
The research investigated the effect of Berberidis Radix, a Tujia medicine, on serum and fecal endogenous metabolites in mice with dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) via metabolomics, aiming to unravel the underlying metabolic pathways and mechanism of action in managing UC. The UC model in mice was generated by the application of DSS. Measurements of body weight, disease activity index (DAI), and colon length were documented. To ascertain the levels of tumor necrosis factor-(TNF-) and interleukin-10(IL-10) in colon tissues, the ELISA technique was utilized. Ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was used to identify and quantify the levels of endogenous metabolites within the serum and feces. landscape genetics Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were applied for the purpose of characterizing and screening differential metabolites. Potential metabolic pathways were subject to analysis by the software MetaboAnalyst 50. Berberidis Radix demonstrably enhanced the well-being of UC mice, exhibiting a noteworthy uptick in anti-inflammatory interleukin-10 (IL-10). From the analysis of serum and fecal samples, 56 differential metabolites, encompassing lipids, amino acids, and fatty acids, were detected in the serum, and 43 in the feces. The metabolic disorder experienced a gradual restoration of function after treatment with Berberidis Radix. Metabolic processes under consideration involved the biosynthesis of phenylalanine, tyrosine, and tryptophan, the metabolism of linoleic acid, the catabolism of phenylalanine, and the metabolism of glycerophospholipids. The observed reduction in DSS-induced ulcerative colitis symptoms in mice treated with Berberidis Radix potentially depends on its modulation of lipid, amino acid, and energy metabolism.
Qualitative and quantitative analysis of 2-(2-phenylethyl) chromones within Aquilaria sinensis suspension cells subjected to sodium chloride (NaCl) treatment was achieved using UPLC-Q-Exactive-MS and UPLC-QQQ-MS/MS. Using a Waters T3 column (21 mm x 50 mm, 18 µm), gradient elution was applied for both analyses, utilizing 0.1% formic acid aqueous solution (A) and acetonitrile (B) as mobile phases. MS data acquisition employed electrospray ionization in positive ion mode. The analysis of NaCl-treated A. sinensis suspension cell samples by UPLC-Q-Exactive-MS identified 47 phenylethylchromones. These comprised 22 flindersia-type 2-(2-phenylethyl) chromones and their glycosides, 10 56,78-tetrahydro-2-(2-phenylethyl) chromones, and a further 15 mono-epoxy or diepoxy-56,78-tetrahydro-2-(2-phenylethyl) chromones. Twenty-five phenylethylchromones were also measured using UPLC-QQQ-MS/MS analysis.