A metabolic exploration of the protective effect of Ligusticum wallichii on IL-1ß-injured mouse chondrocytes.

BACKGROUND: Osteoarthritis (OA) is a metabolic disorder and able to be relieved by traditional Chinese medicines. However, the effect of Ligusticum wallichii on OA is unknown. METHODS: Cytokine IL-1ß and L. wallichii extracts were used to stimulate the primary mouse chondrocytes. MTT assay was used to measure the cell viability. The mRNA and protein level of each gene were test by qRT-PCR and western blotting, respectively. The rate of apoptotic cell was measured by flow cytometry. GC/MS-based metabolomics was utilized to characterize the variation of metabolome. RESULTS: Here, we found that L. wallichii attenuated the IL-1ß-induced apoptosis, inflammatory response, and extracellular matrix (ECM) degradation in mouse chondrocytes. Then we used GC/MS-based metabolomics to characterize the variation of metabolomes. The established metabolic profile of mouse chondrocytes showed that the abundance of most metabolites (n = 40) altered by IL-1ß stimulation could be repressed by L. wallichii treatment. Multivariate data analysis identified that cholesterol, linoleic acid, hexadecandioic acid, proline, l-valine, l-leucine, pyruvate, palmitic acid, and proline are the most key biomarkers for understanding the metabolic role of L. wallichii in IL-1ß-treated chondrocytes. Further pathway analysis using these metabolites enriched fourteen metabolic pathways, which were dramatically changed in IL-1ß-treated chondrocytes and capable of being reprogrammed by L. wallichii incubation. These enriched pathways were involved in carbon metabolisms, fatty acid biosynthesis, and amino acid metabolisms. CONCLUSIONS: These findings provide potential clues that metabolic strategies are linked to protective mechanisms of L. wallichii treatment in IL-1ß-stimulated chondrocytes and emphasize the importance of metabolic strategies against inflammatory responses in OA development.

Separation and indirect ultraviolet detection of ferrous and trivalent iron ions by using ionic liquids in ion chromatography.

A method of simultaneous separation and indirect ultraviolet detection of different valence iron ions Fe2+ and Fe3+ by using ionic liquids as mobile phase additives and ultraviolet absorption reagents on a cation exchange column functionalized with carboxylic acid group was developed. The effects of ionic liquids, organic acids, detection wavelength, etc. on separation and detection of Fe2+ and Fe3+ were investigated and the mechanism was discussed. The pyridinium and imidazolium ionic liquids were not only ultraviolet absorption reagents of indirect ultraviolet detection but also effective components for separating Fe2+ and Fe3+ . The separation and detection of Fe2+ and Fe3+ can be achieved using 0.5 mmol/L pyridinium ionic liquid-1.2 mmol/L methanesulfonic acid as the mobile phase. The determination of Fe2+ and Fe3+ had a good linear relationship in the concentration range of 1-100 mg/L. The limits of detection of Fe2+ and Fe3+ were 0.12 and 0.09 mg/L, respectively. This method was applied to the actual sample detection in the field of medical analysis. The spiked recoveries were between 97.3 and 99.5%, and the relative standard deviations were less than 0.6%. The method is simple, accurate, and reliable, and is an analytical method with universal and practical value.

Overexpression of SCARA5 inhibits tumor proliferation and invasion in osteosarcoma via suppression of the FAK signaling pathway.

Scavenger receptor class A, member 5 (SCARA5) is a member of the scavenger receptor family, and is involved in several types of human malignancy; however, its roles in osteosarcoma (OS) remain to be fully elucidated. Therefore, in the present study, the biological functions of SCARA5 in OS, and the potential underlying mechanisms were investigated. SCARA5 expression in OS tissues and cell lines was detected by reverse transcription­quantitative polymerase chain reaction and western blot analysis. The effects of SCARA5 on the proliferation and migration/invasion ability of OS cells were determined by MTT and Transwell chamber assays, respectively. Expression levels of phosphorylated focal adhesion kinase (p­FAK), FAK, p­Src, Src, matrix metalloproteinase (MMP)2 and MMP9 were evaluated via western blot analysis. The results of the present study demonstrated that SCARA5 was expressed at low levels in OS tissues and cell lines. The overexpression of SCARA5 significantly inhibited the proliferation, colony formation and migration/invasion abilities of the OS cells. Furthermore, SCARA5 significantly decreased the expression levels of p­FAK, MMP­2 and MMP­9 in the OS cells. Taken together, these data suggested that the overexpression of SCARA5 inhibits tumor proliferation and invasion in OS via suppression of the FAK signaling pathway. Thus, novel therapeutic strategies or drugs targeted at SCARA5 may offer potential for the treatment of OS.