Abstract
BackgroundScutellaria baicalensis Georgi is a commonly used medicinal herb in several Asian countries like Korea, China and Japan for thousands of years. It has been reported to have various medicinal properties such as anti-microbial, anti-inflammatory and anti-cancer effects. However, the anti-inflammatory mechanism of S. baicalensis G at proteome level has not yet been reported. Hence, we performed a proteome analysis to study differentially expressed proteins and its anti-inflammatory role in lipopolysaccharide (LPS) stimulated L6 skeletal muscle cells response to flavonoids isolated from S. baicalensis G.MethodsFor that, 150 μg of proteins from the L6 cells of the control (Vehicle only), LPS treated and flavonoid treated groups were separated using 18 cm, pH 4–7 IPG strips in the first dimension and resolved by 12% linear gradient SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The silver stained gels were analyzed by using progenesis SameSpots software and twenty six differentially expressed protein spots (≥2 fold, p < 0.05) were selected for matrix assisted laser desorption ionization- time of flight mass spectroscopy/mass spectrometry (MALDI-TOF/MS) analysis. Also, the expression of COX-2, iNOS and Annexin A2 proteins were analyzed by western blot.ResultsTotally, 12 differentially expressed proteins were successfully identified by MALDI-TOF/MS and database searching, that’s involved in inflammatory responses such vimentin, T-box transcription factor TBX3, annexin A1, annexin A2 and annexin A5. In addition, flavonoids inhibited the expression of COX-2, iNOS and Annexin A2 proteins in LPS-stimulated L6 skeletal muscle cells.ConclusionsThe findings revealed that the flavonoids from S. baicalensis G. directly protect the LPS stimulated inflammation process in L6 cells and, would be helpful to study further the muscle cell inflammatory mechanism. This is the first proteome study provide the anti-inflammatory mechanism of flavonoids from S. baicalensis G. in LPS stimulated L6 skeletal muscle cells.
Highlights
Scutellaria baicalensis Georgi is a commonly used medicinal herb in several Asian countries like Korea, China and Japan for thousands of years
Quantification and characterization of flavonoids from Scutellaria baicalensis G Flavonoids were isolated from Korean S. bicalensis G. using High performance liquid chromatography (HPLC)-MS/MS. totally 16 peaks were identified based on the HPLC retention time, molecular ion masses and the ultraviolet–visible spectra of standard compounds in a library
In conclusion, we demonstrated that the anti-inflammatory effects of flavonoids from S. baicalensis G. on LPS stimulated L6 cells using proteomic approach
Summary
Scutellaria baicalensis Georgi is a commonly used medicinal herb in several Asian countries like Korea, China and Japan for thousands of years. We performed a proteome analysis to study differentially expressed proteins and its anti-inflammatory role in lipopolysaccharide (LPS) stimulated L6 skeletal muscle cells response to flavonoids isolated from S. baicalensis G. Anti-inflammatory effect of Korean S. baicalensis G. in LPS induced L6 skeletal muscle cells using proteomic approach has not been studied. In the present study, we performed a proteome analysis of L6 cells after treatment with LPS and flavonoids Korean S. baicalensis G. and 12 differentially expressed proteins were identified by MALDI-TOF/MS. These proteome results suggest that flavonoids from S. baicalensis G. directly protect the LPS stimulated inflammation process in L6 skeletal muscle cells. This is the first study provide the evidence for an interaction between flavonoids from S. baicalensis G. and LPS stimulated L6 skeletal muscle cells
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