Global and Targeted Metabolomics Reveal That Bupleurotoxin, a Toxic Type of Polyacetylene, Induces Cerebral Lesion by Inhibiting GABA Receptor in Mice

Polyacetylenes are widely distributed in food plants and medicinal herbs, which have been shown to have highly neurotoxic effects. However, there were insufficient studies on the toxicity of these compounds. Thus, a series of experiments was designed to elucidate the toxicity mechanism of bupleurotoxin (BETX) as a representative polyacetylene. First, male BALB/c mice were intragastrically administered 2.5 mg/kg of bodyweight BETX once a day for seven consecutive days. The histopathological results showed that BETX could induce severe morphological damages in the brain hippocampus. We then used metabolomics approaches to screen serum samples from the control and BETX-treated groups. The global metabolomics results revealed 17 metabolites that were perturbed after BETX treatment. Four of these metabolites were then verified by targeted metabolomics. Bioinformatics analysis with the Ingenuity Pathway Analysis (IPA) software found a strong correlation between the GABA receptor signaling pathway and these metabolites. On the basis of these results, a validation test using a rat hippocampal neuron cell line was performed, and the results confirmed that BETX inhibited GABA-induced currents (<i>I</i>_GABA) in a competitive manner. In summary, our study illustrated the molecular mechanism of the toxicity of polyacetylenes. In addition, our study was instructive for the study of other toxic medical herbs

Multiple compounds improve aged MDSPC proliferation (self-renewal).

<p>(A) Aged MDSPCs were cultured in medium containing aspirin, nordihydroguaiaretic acid (NDGA), or rapamycin, and proliferation was monitored by live cell imaging for up to 60 hours, or (B) cells were treated in fusion medium and assessed for myogenic differentiation after 5 days (*p≤0.05 Aged WT vs Treated; ANOVA followed by Tukey post-hoc analysis).</p

MOESM1 of Application of a strategy based on metabolomics guided promoting blood circulation bioactivity compounds screening of vinegar

Additional file 1: Table S1. The content of TMPZ in RV and WV. Table S2. The peak area and the relative peak area value of four potential biomarkers in different aging period. Table S3. The levels and factors investigated in BBD. Figure S1. HPLC chromatogram of TMPZ. Figure S2. The results of bioactivity screening. Figure S3. Diagnostic efficacy evaluation using ROC curves of the four potential biomarker metabolites in two different vinegar. Figure S4. Trends of time-series analysis graphs of four potential biomarkers. (A) TMPZ (MAPE: 2.05853, MAD: 1.67627, fitted curve: Yt = 60.81+5.089xt); (B) Dihydroergotamine (MAPE: 1.63096, MAD: 0.15345, fitted curve: Yt = 6.726+0.7121xt); (C) Harmine (MAPE: 1.72704, MAD: 0.01711, fitted curve: Yt = 0.7764+0.05780xt); (D) 1,2,3,4-tetrahydroharmine (MAPE: 3.76071, MAD: 0.04998, fitted curve: Yt = 0.9695+0.0910xt). Figure S5. Response surfaces estimated from the full factorial design for the content of total alkaloids

NF-κB inhibition decreases senescence in BM-MSCs.

<p>BM-MSCs were treated for 48 hours with IKK inhibitor VII and then stained for senescence-associated beta-galactosidase (SA β-gal) expression, which demonstrated a decrease in cell senescence (*p≤0.05, T-test).</p

NF-κB inhibition improves the differentiation of MDSPCs isolated from old mice.

<p>(A) Brightfield images demonstrate a dose dependent increase in myotube formation following IKKi administration (10x images). (B) Immunofluorescent staining for MyHC demonstrated significantly improved differentiation following treatment at the 5μM dose (20x magnification), quantified in (C). (D) The total number of nuclei per field of view for experiment in B, C (*p≤0.05; ANOVA followed by Tukey post-hoc analysis). (E) IKKi treatment of young WT MDSPCs did not significantly improve differentiation (*p≤0.05, T-test).</p

The Shared Crosstalk of Multiple Pathways Involved in the Inflammation between Rheumatoid Arthritis and Coronary Artery Disease Based on a Digital Gene Expression Profile

<div><p>Rheumatoid arthritis (RA) and coronary artery disease (CAD) are both complex inflammatory diseases, and an increased prevalence of CAD and a high rate of mortality have been observed in RA patients. But the molecular mechanism of inflammation that is shared between the two disorders is unclear. High-throughput techniques, such as transcriptome analysis, are becoming important tools for genetic biomarker discovery in highly complex biological samples, which is critical for the diagnosis, prognosis, and treatment of disease. In the present study, we reported one type of transcriptome analysis method: digital gene expression profiling of peripheral blood mononuclear cells of 10 RA patients, 10 CAD patients and 10 healthy people. In all, 213 and 152 differently expressed genes (DEGs) were identified in RA patients compared with normal controls (RA <i>vs.</i> normal) and CAD patients compared with normal controls (CAD <i>vs.</i> normal), respectively, with 73 shared DEGs between them. Using this technique in combination with Ingenuity Pathways Analysis software, the effects on inflammation of four shared canonical pathways, three shared activated predicted upstream regulators and three shared molecular interaction networks were identified and explored. These shared molecular mechanisms may provide the genetic basis and potential targets for optimizing the application of current drugs to more effectively treat these diseases simultaneously and for preventing one when the other is diagnosed.</p></div

Additional file 2: Table S1. of Virtual screening of the inhibitors targeting at the viral protein 40 of Ebola virus

List of top 15 compounds from TCM database. Table S2. Top ranking TCM lead compounds obtained using iScreen against VP40. Table S3. Comparison of binding free energies of lead compounds using Autodock. Table S4. Energetic contribution of lead compounds and RNA with VP40. Table S5. Comparison of interacting residues of lead compounds. Table S6. ADME and Drug-likeness analysis. Table S7. Toxicity risks of lead compounds predicted by OSIRIS property explorer. Table S8. Prediction of oral toxicity (LD50). Figure S1. Overall flow chart of the present study. Figure S2. Crucial Hydrogen bond interaction formed between RNA and VP40. Figure S3. Crucial Hydrogen bond interaction formed between compound1 and VP40. Figure S4. Crucial Hydrogen bond interaction formed between compund2 and VP40. (DOC 8072 kb

NF-κB inhibition improves the oxidative stress resistance of MDSPCs isolated from old mice.

<p>MDSPCs were exposed to 250 μM H₂O₂ and monitored by live cell imaging for up to 24 hours. IKKi treated aged WT MDSPCs and aged <i>p65</i>^+/- MDSPCs demonstrated improved survival compared to vehicle treated aged WT MDSPCs (*p≤0.05 Aged WT vs +IKKi; +p≤0.05 Aged WT vs Aged <i>p65</i>^+/-; ANOVA followed by Tukey post-hoc analysis).</p

Network, molecular functions and canonical pathway analysis of shared DEGs in RA <i>vs.</i> normal and CAD <i>vs</i>. normal individuals.

<p>A: The molecular network of 95 shared DEGs in RA <i>vs.</i> normal and CAD <i>vs.</i> normal individuals; blue molecules represent the shared DEGs; B: the top 4 shared canonical pathways (with score (-log (<i>p</i>-value))>1.3, <i>p</i>-value of pathway <0.05) related with inflammation which 95 shared DEGs in RA vs. normal and CAD vs. normal involved in: IL-8 signaling; antigen presentation pathway; OX40 signaling pathway; TREM1 signaling; C: the function classification of shared DEGs in both RA <i>vs.</i> normal and CAD <i>vs.</i> normal, the number in the bar diagram represented the DEGs numbers participated in the corresponding pathways.</p

The top 4 shared signaling pathways related to inflammation and shared DEGs in both RA <i>vs</i>. normal and CAD <i>vs</i>. normal subjects.

<p>The top 4 shared signaling pathways related to inflammation and shared DEGs in both RA <i>vs</i>. normal and CAD <i>vs</i>. normal subjects.</p