Project description:Hypericum perforatum extracts have been used as dietary supplements to treat conditions including mild-moderate depression and inflammation. A group of four bioactive constituents were identified from an active fraction of the extract. In order to identify the mechanism for the potential anti-inflammatory activity of the identified compounds, we used Affymatrix microarray to study the gene expression profile impacteded by these compounds, as well as the active fraction in LPS-stimulated mouse macrophages. We treated RAW264.7 mouse macrophages with DMSO control, active fraction from Hypericum perforaum extract, and a combination of the 4 putative bioactive compounds, called the 4-component system, all with and without LPS induction. A total of six treatment combinations were included in the final gene expression analysis using microarray.
Project description:Hypericum perforatum extracts have been used as dietary supplements to treat conditions including mild-moderate depression and inflammation. A group of four bioactive constituents were identified from an active fraction of the extract. In order to identify the mechanism for the potential anti-inflammatory activity of the identified compounds, we used Affymatrix microarray to study the gene expression profile impacteded by these compounds, as well as the active fraction in LPS-stimulated mouse macrophages.
Project description:To screen for altered gene expression during osteoclastogenesis, BMM cells treated with RANKL or RANKL+LEA were subjected to gene expression profiling.
Project description:The success of bottom-up proteomic analysis frequently depends on the efficient removal of contaminants from protein or peptide samples before LC-MS/MS. For a peptide clean-up workflow, the single-pot solid-phase-enhanced peptide sample preparation on carboxylate-modified paramagnetic beads (termed SP2) was evaluated for sodium dodecyl sulfate or polyethylene glycol removal from Arabidopsis thaliana tryptic peptides. The robust and efficient 40-min SP2 protocol, tested for a 10 ng, 250ng and 10µg peptide sample, was proposed and benchmarked thoroughly against the ethyl acetate extraction protocol. The SP2 protocol on carboxylated magnetic beads proved to be the robust approach even for simultaneous removal of massive sodium dodecyl sulfate (SDS) and polyethylene glycol (PEG) contaminations from AT peptide samples in respect of the LC-MS/MS data outperforming ethyl acetate extraction.
Project description:Sargassum micracanthum, a brown macroalga rich in bioactive metabolites, has been widely recognized for its pharmacological potential. However, its role in collagen production and extracellular matrix (ECM) remodeling for skin regeneration remains largely unexplored. Therefore, this study examined the effects of S. micracanthum ethanolic extract on collagen-related pathways in human dermal fibroblasts (HDFs) through RNA sequencing and in vitro analysis. A total of 657 differentially expressed genes were identified, including 15 associated with collagen synthesis. Notably, COL4A1, COL4A2, COL5A3, and COL18A1 were significantly upregulated in the extract-treated group. Gene ontology analysis revealed enrichment in collagen-related terms, while WikiPathway analysis indicated upregulation of growth-factor-related pathways, which are crucial for cell proliferation, ECM remodeling, and tissue regeneration. In vitro experiments demonstrated a dose-dependent increase in collagen production, with a significant enhancement observed at 100 μg/mL (p < 0.05). These findings demonstrate the potential applicability of S. micracanthum in skin regeneration and biomedical therapies.