Limits...
Protective effects of andrographolide analogue AL-1 on ROS-induced RIN-mβ cell death by inducing ROS generation.

Yan GR, Zhou HH, Wang Y, Zhong Y, Tan ZL, Wang Y, He QY - PLoS ONE (2013)

Bottom Line: In this work, we used proteomics to identify AL-1-regulated proteins in β-cells and found that 13 of the 71 proteins regulated by AL-1 were closely associated with antioxidation.Functional investigation demonstrated that AL-1 exerted its protective effects on H2O2-induced cell death of β-cells by generating NADPH oxidase-dependent ROS to activate ERK1/2 and AKT1 signaling pathways.To the best of our knowledge, this is the first comprehensive proteomic analysis illustrating a novel molecular mechanism for the protective effects of antioxidants on β-cells from H2O2-induced cell death.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University, Guangzhou, China. tgryan@jnu.edu.cn

ABSTRACT
Oxidative stress is considered to be a major factor contributing to pathogenesis and progression of many diseases. A novel andrographolide-lipoic acid conjugate (AL-1) could protect pancreatic β-cells from reactive oxygen species (ROS)-induced oxidative injury. However, its protective mechanism is still unclear. In this work, we used proteomics to identify AL-1-regulated proteins in β-cells and found that 13 of the 71 proteins regulated by AL-1 were closely associated with antioxidation. These differential proteins were mainly involved in the ERK1/2 and AKT1 signaling pathways. Functional investigation demonstrated that AL-1 exerted its protective effects on H2O2-induced cell death of β-cells by generating NADPH oxidase-dependent ROS to activate ERK1/2 and AKT1 signaling pathways. As a consequence, the expressions of antioxidant proteins including Trx1, Prx1 and Prx5, and anti-apoptotic proteins including PDCD6IP, prohibitin, galectin-1 and HSP were upregulated. AL-1 probably worked as a "vaccinum" to activate the cellular antioxidant system by inducing the generation of low concentration ROS which then reciprocally protected β-cells from oxidative damage caused by high-level ROS from H2O2. To the best of our knowledge, this is the first comprehensive proteomic analysis illustrating a novel molecular mechanism for the protective effects of antioxidants on β-cells from H2O2-induced cell death.

Show MeSH

Related in: MedlinePlus

Image overview of 2-DE gels for the proteins extracted from RIN-mβ cells treated with and without AL-1 for 1 h.150 μg proteins from RIN-mβ cells treated with and without AL-1 were separated by 2-DE, and the gels were stained with sliver. Shown are the representative results from three independent experiments.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3672203&req=5

pone-0063656-g002: Image overview of 2-DE gels for the proteins extracted from RIN-mβ cells treated with and without AL-1 for 1 h.150 μg proteins from RIN-mβ cells treated with and without AL-1 were separated by 2-DE, and the gels were stained with sliver. Shown are the representative results from three independent experiments.

Mentions: Total proteins extracted from RIN-mβ cells treated with and without 0.1 μM AL-1 for 1 h were separated on 2-DE to compare the differential proteins regulated by AL-1 (Fig. 2). Altogether, Over 1000 protein spots were detected in each gel by using ImageMaster software. Protein spots altered greater than 1.5-fold in spot intensity and observed in three replicate gels from three independent experiments were scored and subjected to MS analysis. This allowed us to identify 71 proteins from 105 reproducible differential spots, including 52 increases and 19 decreases in AL-1 treatment gels (Table S1).


Protective effects of andrographolide analogue AL-1 on ROS-induced RIN-mβ cell death by inducing ROS generation.

Yan GR, Zhou HH, Wang Y, Zhong Y, Tan ZL, Wang Y, He QY - PLoS ONE (2013)

Image overview of 2-DE gels for the proteins extracted from RIN-mβ cells treated with and without AL-1 for 1 h.150 μg proteins from RIN-mβ cells treated with and without AL-1 were separated by 2-DE, and the gels were stained with sliver. Shown are the representative results from three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3672203&req=5

pone-0063656-g002: Image overview of 2-DE gels for the proteins extracted from RIN-mβ cells treated with and without AL-1 for 1 h.150 μg proteins from RIN-mβ cells treated with and without AL-1 were separated by 2-DE, and the gels were stained with sliver. Shown are the representative results from three independent experiments.
Mentions: Total proteins extracted from RIN-mβ cells treated with and without 0.1 μM AL-1 for 1 h were separated on 2-DE to compare the differential proteins regulated by AL-1 (Fig. 2). Altogether, Over 1000 protein spots were detected in each gel by using ImageMaster software. Protein spots altered greater than 1.5-fold in spot intensity and observed in three replicate gels from three independent experiments were scored and subjected to MS analysis. This allowed us to identify 71 proteins from 105 reproducible differential spots, including 52 increases and 19 decreases in AL-1 treatment gels (Table S1).

Bottom Line: In this work, we used proteomics to identify AL-1-regulated proteins in β-cells and found that 13 of the 71 proteins regulated by AL-1 were closely associated with antioxidation.Functional investigation demonstrated that AL-1 exerted its protective effects on H2O2-induced cell death of β-cells by generating NADPH oxidase-dependent ROS to activate ERK1/2 and AKT1 signaling pathways.To the best of our knowledge, this is the first comprehensive proteomic analysis illustrating a novel molecular mechanism for the protective effects of antioxidants on β-cells from H2O2-induced cell death.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University, Guangzhou, China. tgryan@jnu.edu.cn

ABSTRACT
Oxidative stress is considered to be a major factor contributing to pathogenesis and progression of many diseases. A novel andrographolide-lipoic acid conjugate (AL-1) could protect pancreatic β-cells from reactive oxygen species (ROS)-induced oxidative injury. However, its protective mechanism is still unclear. In this work, we used proteomics to identify AL-1-regulated proteins in β-cells and found that 13 of the 71 proteins regulated by AL-1 were closely associated with antioxidation. These differential proteins were mainly involved in the ERK1/2 and AKT1 signaling pathways. Functional investigation demonstrated that AL-1 exerted its protective effects on H2O2-induced cell death of β-cells by generating NADPH oxidase-dependent ROS to activate ERK1/2 and AKT1 signaling pathways. As a consequence, the expressions of antioxidant proteins including Trx1, Prx1 and Prx5, and anti-apoptotic proteins including PDCD6IP, prohibitin, galectin-1 and HSP were upregulated. AL-1 probably worked as a "vaccinum" to activate the cellular antioxidant system by inducing the generation of low concentration ROS which then reciprocally protected β-cells from oxidative damage caused by high-level ROS from H2O2. To the best of our knowledge, this is the first comprehensive proteomic analysis illustrating a novel molecular mechanism for the protective effects of antioxidants on β-cells from H2O2-induced cell death.

Show MeSH
Related in: MedlinePlus