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Proteome profiling of lipopolysaccharide induced L6 rat skeletal muscle cells response to flavonoids from Scutellaria baicalensis Georgi.

Kim JA, Nagappan A, Park HS, Saralamma VV, Hong GE, Yumnam S, Lee HJ, Raha S, Kim EH, Young PS, Kim GS - BMC Complement Altern Med (2014)

Bottom Line: It has been reported to have various medicinal properties such as anti-microbial, anti-inflammatory and anti-cancer effects.Totally, 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.This is the first proteome study provide the anti-inflammatory mechanism of flavonoids from S. baicalensis G. in LPS stimulated L6 skeletal muscle cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul 137-701, Republic of Korea. paik@catholic.ac.kr.

ABSTRACT

Background: Scutellaria 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.

Methods: For 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.

Results: Totally, 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.

Conclusions: The 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.

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Related in: MedlinePlus

Effects of flavonoids on lipopolysaccharide induced L6 skeletal muscle cells. L6 cells were untreated or treated with the indicated concentration of LPS and flavonoids for 24 h, and COX-2, iNOS and Annexin A2 protein levels in the cell lysates were assayed by Western blot analysis. The experiments were conducted at least three replications.
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Fig4: Effects of flavonoids on lipopolysaccharide induced L6 skeletal muscle cells. L6 cells were untreated or treated with the indicated concentration of LPS and flavonoids for 24 h, and COX-2, iNOS and Annexin A2 protein levels in the cell lysates were assayed by Western blot analysis. The experiments were conducted at least three replications.

Mentions: The effect of flavonoids on the expression of COX-2 and iNOS proteins were determined by western blot. In LPS-stimulated L6 skeletal muscle cells, the expression of COX-2 and iNOS proteins were remarkably increased and pretreatment with flavonoid (150 μg/mL) attenuated protein expression (Figure 4). These results were suggest that flavonoids can inhibit LPS-induced inflammation by suppression COX-2 and iNOS expression at translational levels. Moreover, an anti-inflammatory protein, Anxa2 (unique annexins) expression was increased in LPS-treated cells and, inhibited by flavonoids in LPS stimulated L6 skeletal muscle cells (Figures 4 and 5D). These results consistent with 2-DE results that LPS can induce inflammation in L6 cells by increasing Anxa2 protein and, that can be inhibited by flavonoids which isolated from S. baicalensis G.Figure 4


Proteome profiling of lipopolysaccharide induced L6 rat skeletal muscle cells response to flavonoids from Scutellaria baicalensis Georgi.

Kim JA, Nagappan A, Park HS, Saralamma VV, Hong GE, Yumnam S, Lee HJ, Raha S, Kim EH, Young PS, Kim GS - BMC Complement Altern Med (2014)

Effects of flavonoids on lipopolysaccharide induced L6 skeletal muscle cells. L6 cells were untreated or treated with the indicated concentration of LPS and flavonoids for 24 h, and COX-2, iNOS and Annexin A2 protein levels in the cell lysates were assayed by Western blot analysis. The experiments were conducted at least three replications.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4195865&req=5

Fig4: Effects of flavonoids on lipopolysaccharide induced L6 skeletal muscle cells. L6 cells were untreated or treated with the indicated concentration of LPS and flavonoids for 24 h, and COX-2, iNOS and Annexin A2 protein levels in the cell lysates were assayed by Western blot analysis. The experiments were conducted at least three replications.
Mentions: The effect of flavonoids on the expression of COX-2 and iNOS proteins were determined by western blot. In LPS-stimulated L6 skeletal muscle cells, the expression of COX-2 and iNOS proteins were remarkably increased and pretreatment with flavonoid (150 μg/mL) attenuated protein expression (Figure 4). These results were suggest that flavonoids can inhibit LPS-induced inflammation by suppression COX-2 and iNOS expression at translational levels. Moreover, an anti-inflammatory protein, Anxa2 (unique annexins) expression was increased in LPS-treated cells and, inhibited by flavonoids in LPS stimulated L6 skeletal muscle cells (Figures 4 and 5D). These results consistent with 2-DE results that LPS can induce inflammation in L6 cells by increasing Anxa2 protein and, that can be inhibited by flavonoids which isolated from S. baicalensis G.Figure 4

Bottom Line: It has been reported to have various medicinal properties such as anti-microbial, anti-inflammatory and anti-cancer effects.Totally, 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.This is the first proteome study provide the anti-inflammatory mechanism of flavonoids from S. baicalensis G. in LPS stimulated L6 skeletal muscle cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul 137-701, Republic of Korea. paik@catholic.ac.kr.

ABSTRACT

Background: Scutellaria 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.

Methods: For 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.

Results: Totally, 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.

Conclusions: The 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.

Show MeSH
Related in: MedlinePlus