Lycium barbarum polysaccharides protect human lens epithelial cells against oxidative stress-induced apoptosis and senescence
We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress-induced apoptosis and senescence in human lens epithelial cells. To study apoptosis, SRA01/04 cells, a human lens epithelial cell lines, were exposed to 200 µM hydrogen peroxide (H2O2) fo...
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| Published in | PloS one Vol. 9; no. 10; p. e110275 |
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| Main Authors | , , , , , , , |
| Format | Journal Article |
| Language | English |
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United States
Public Library of Science
15.10.2014
Public Library of Science (PLoS) |
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| Abstract | We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress-induced apoptosis and senescence in human lens epithelial cells.
To study apoptosis, SRA01/04 cells, a human lens epithelial cell lines, were exposed to 200 µM hydrogen peroxide (H2O2) for 24 h with or without pretreatment with LBPs. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis, intracellular reactive oxygen species (ROS), and the loss of mitochondria membrane potential (Δψm) were detected by flow cytometric analyses. Expression levels of Bcl-2 and Bax proteins were measured by western blot analysis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) were quantized using commercial enzymatic kits according to the manufacturer's instructions. To study senescence, SRA01/04 cells were pre-incubated with LBPs and all cells were then exposed to 100 µM H2O2 for 96 h. Cellular senescence was assessed by morphologic examination and senescence-associated β-galactosidase (SA-β-gal) staining.
LBPs significantly reduced H2O2-induced cell apoptosis, the generation of ROS, the loss of Δψm, and the levels of MDA. LBPs also inhibited H2O2-induced downregulated Bcl-2 and upregulated Bax proteins and increased the levels of SOD and GSH enzyme activity. Moreover, LBPs significantly attenuated H2O2-induced cellular senescence.
These findings suggested that LBPs protect human lens epithelial cells from H2O2-induced apoptosis by modulating the generation of ROS, loss of Δψm, Bcl-2 family, and antioxidant enzyme activity and attenuating cellular senescence. |
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| AbstractList | We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress-induced apoptosis and senescence in human lens epithelial cells. To study apoptosis, SRA01/04 cells, a human lens epithelial cell lines, were exposed to 200 [micro]M hydrogen peroxide (H.sub.2 O.sub.2) for 24 h with or without pretreatment with LBPs. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis, intracellular reactive oxygen species (ROS), and the loss of mitochondria membrane potential ([DELTA][psi]m) were detected by flow cytometric analyses. Expression levels of Bcl-2 and Bax proteins were measured by western blot analysis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) were quantized using commercial enzymatic kits according to the manufacturer's instructions. To study senescence, SRA01/04 cells were pre-incubated with LBPs and all cells were then exposed to 100 [micro]M H.sub.2 O.sub.2 for 96 h. Cellular senescence was assessed by morphologic examination and senescence-associated [beta]-galactosidase (SA-[beta]-gal) staining. LBPs significantly reduced H.sub.2 O.sub.2 -induced cell apoptosis, the generation of ROS, the loss of [DELTA][psi]m, and the levels of MDA. LBPs also inhibited H.sub.2 O.sub.2 -induced downregulated Bcl-2 and upregulated Bax proteins and increased the levels of SOD and GSH enzyme activity. Moreover, LBPs significantly attenuated H.sub.2 O.sub.2 -induced cellular senescence. These findings suggested that LBPs protect human lens epithelial cells from H.sub.2 O.sub.2 -induced apoptosis by modulating the generation of ROS, loss of [DELTA][psi]m, Bcl-2 family, and antioxidant enzyme activity and attenuating cellular senescence. We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress-induced apoptosis and senescence in human lens epithelial cells.To study apoptosis, SRA01/04 cells, a human lens epithelial cell lines, were exposed to 200 µM hydrogen peroxide (H2O2) for 24 h with or without pretreatment with LBPs. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis, intracellular reactive oxygen species (ROS), and the loss of mitochondria membrane potential (Δψm) were detected by flow cytometric analyses. Expression levels of Bcl-2 and Bax proteins were measured by western blot analysis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) were quantized using commercial enzymatic kits according to the manufacturer's instructions. To study senescence, SRA01/04 cells were pre-incubated with LBPs and all cells were then exposed to 100 µM H2O2 for 96 h. Cellular senescence was assessed by morphologic examination and senescence-associated β-galactosidase (SA-β-gal) staining.LBPs significantly reduced H2O2-induced cell apoptosis, the generation of ROS, the loss of Δψm, and the levels of MDA. LBPs also inhibited H2O2-induced downregulated Bcl-2 and upregulated Bax proteins and increased the levels of SOD and GSH enzyme activity. Moreover, LBPs significantly attenuated H2O2-induced cellular senescence.These findings suggested that LBPs protect human lens epithelial cells from H2O2-induced apoptosis by modulating the generation of ROS, loss of Δψm, Bcl-2 family, and antioxidant enzyme activity and attenuating cellular senescence. Objectives We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress-induced apoptosis and senescence in human lens epithelial cells. Methods To study apoptosis, SRA01/04 cells, a human lens epithelial cell lines, were exposed to 200 [micro]M hydrogen peroxide (H.sub.2 O.sub.2) for 24 h with or without pretreatment with LBPs. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis, intracellular reactive oxygen species (ROS), and the loss of mitochondria membrane potential ([DELTA][psi]m) were detected by flow cytometric analyses. Expression levels of Bcl-2 and Bax proteins were measured by western blot analysis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) were quantized using commercial enzymatic kits according to the manufacturer's instructions. To study senescence, SRA01/04 cells were pre-incubated with LBPs and all cells were then exposed to 100 [micro]M H.sub.2 O.sub.2 for 96 h. Cellular senescence was assessed by morphologic examination and senescence-associated [beta]-galactosidase (SA-[beta]-gal) staining. Results LBPs significantly reduced H.sub.2 O.sub.2 -induced cell apoptosis, the generation of ROS, the loss of [DELTA][psi]m, and the levels of MDA. LBPs also inhibited H.sub.2 O.sub.2 -induced downregulated Bcl-2 and upregulated Bax proteins and increased the levels of SOD and GSH enzyme activity. Moreover, LBPs significantly attenuated H.sub.2 O.sub.2 -induced cellular senescence. Conclusions These findings suggested that LBPs protect human lens epithelial cells from H.sub.2 O.sub.2 -induced apoptosis by modulating the generation of ROS, loss of [DELTA][psi]m, Bcl-2 family, and antioxidant enzyme activity and attenuating cellular senescence. We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress-induced apoptosis and senescence in human lens epithelial cells. To study apoptosis, SRA01/04 cells, a human lens epithelial cell lines, were exposed to 200 µM hydrogen peroxide (H2O2) for 24 h with or without pretreatment with LBPs. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis, intracellular reactive oxygen species (ROS), and the loss of mitochondria membrane potential (Δψm) were detected by flow cytometric analyses. Expression levels of Bcl-2 and Bax proteins were measured by western blot analysis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) were quantized using commercial enzymatic kits according to the manufacturer's instructions. To study senescence, SRA01/04 cells were pre-incubated with LBPs and all cells were then exposed to 100 µM H2O2 for 96 h. Cellular senescence was assessed by morphologic examination and senescence-associated β-galactosidase (SA-β-gal) staining. LBPs significantly reduced H2O2-induced cell apoptosis, the generation of ROS, the loss of Δψm, and the levels of MDA. LBPs also inhibited H2O2-induced downregulated Bcl-2 and upregulated Bax proteins and increased the levels of SOD and GSH enzyme activity. Moreover, LBPs significantly attenuated H2O2-induced cellular senescence. These findings suggested that LBPs protect human lens epithelial cells from H2O2-induced apoptosis by modulating the generation of ROS, loss of Δψm, Bcl-2 family, and antioxidant enzyme activity and attenuating cellular senescence. Objectives We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress–induced apoptosis and senescence in human lens epithelial cells. Methods To study apoptosis, SRA01/04 cells, a human lens epithelial cell lines, were exposed to 200 µM hydrogen peroxide (H2O2) for 24 h with or without pretreatment with LBPs. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis, intracellular reactive oxygen species (ROS), and the loss of mitochondria membrane potential (Δψm) were detected by flow cytometric analyses. Expression levels of Bcl-2 and Bax proteins were measured by western blot analysis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) were quantized using commercial enzymatic kits according to the manufacturer's instructions. To study senescence, SRA01/04 cells were pre-incubated with LBPs and all cells were then exposed to 100 µM H2O2 for 96 h. Cellular senescence was assessed by morphologic examination and senescence-associated β-galactosidase (SA-β-gal) staining. Results LBPs significantly reduced H2O2-induced cell apoptosis, the generation of ROS, the loss of Δψm, and the levels of MDA. LBPs also inhibited H2O2-induced downregulated Bcl-2 and upregulated Bax proteins and increased the levels of SOD and GSH enzyme activity. Moreover, LBPs significantly attenuated H2O2-induced cellular senescence. Conclusions These findings suggested that LBPs protect human lens epithelial cells from H2O2-induced apoptosis by modulating the generation of ROS, loss of Δψm, Bcl-2 family, and antioxidant enzyme activity and attenuating cellular senescence. Objectives We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress–induced apoptosis and senescence in human lens epithelial cells. Methods To study apoptosis, SRA01/04 cells, a human lens epithelial cell lines, were exposed to 200 µM hydrogen peroxide (H 2 O 2 ) for 24 h with or without pretreatment with LBPs. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis, intracellular reactive oxygen species (ROS), and the loss of mitochondria membrane potential (Δψm) were detected by flow cytometric analyses. Expression levels of Bcl-2 and Bax proteins were measured by western blot analysis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) were quantized using commercial enzymatic kits according to the manufacturer's instructions. To study senescence, SRA01/04 cells were pre-incubated with LBPs and all cells were then exposed to 100 µM H 2 O 2 for 96 h. Cellular senescence was assessed by morphologic examination and senescence-associated β-galactosidase (SA-β-gal) staining. Results LBPs significantly reduced H 2 O 2 -induced cell apoptosis, the generation of ROS, the loss of Δψm, and the levels of MDA. LBPs also inhibited H 2 O 2 -induced downregulated Bcl-2 and upregulated Bax proteins and increased the levels of SOD and GSH enzyme activity. Moreover, LBPs significantly attenuated H 2 O 2 -induced cellular senescence. Conclusions These findings suggested that LBPs protect human lens epithelial cells from H 2 O 2 -induced apoptosis by modulating the generation of ROS, loss of Δψm, Bcl-2 family, and antioxidant enzyme activity and attenuating cellular senescence. |
| Audience | Academic |
| Author | Wang, Guifang Guo, Xiaoling Wen, Yuechun Hou, Guanghui Liu, Lian Qi, Bing Zhong, Jingxiang Ji, Qingshan |
| AuthorAffiliation | 3 Department of Cell Biology, the Cell Biology Institute of Jinan University, Guangzhou, Guangdong, China 1 Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China University of Pecs Medical School, Hungary 2 Department of Ophthalmology, Affiliated Anhui Provincial Hospital of Anhui Medical University, Hefei, Anhui, China 4 Department of Ophthalmology, the Third Affiliated Hospital of Jinan University, Zhuhai, Guangdong, China |
| AuthorAffiliation_xml | – name: 1 Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China – name: 3 Department of Cell Biology, the Cell Biology Institute of Jinan University, Guangzhou, Guangdong, China – name: University of Pecs Medical School, Hungary – name: 2 Department of Ophthalmology, Affiliated Anhui Provincial Hospital of Anhui Medical University, Hefei, Anhui, China – name: 4 Department of Ophthalmology, the Third Affiliated Hospital of Jinan University, Zhuhai, Guangdong, China |
| Author_xml | – sequence: 1 givenname: Bing surname: Qi fullname: Qi, Bing organization: Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China – sequence: 2 givenname: Qingshan surname: Ji fullname: Ji, Qingshan organization: Department of Ophthalmology, Affiliated Anhui Provincial Hospital of Anhui Medical University, Hefei, Anhui, China – sequence: 3 givenname: Yuechun surname: Wen fullname: Wen, Yuechun organization: Department of Ophthalmology, Affiliated Anhui Provincial Hospital of Anhui Medical University, Hefei, Anhui, China – sequence: 4 givenname: Lian surname: Liu fullname: Liu, Lian organization: Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China – sequence: 5 givenname: Xiaoling surname: Guo fullname: Guo, Xiaoling organization: Department of Cell Biology, the Cell Biology Institute of Jinan University, Guangzhou, Guangdong, China – sequence: 6 givenname: Guanghui surname: Hou fullname: Hou, Guanghui organization: Department of Ophthalmology, the Third Affiliated Hospital of Jinan University, Zhuhai, Guangdong, China – sequence: 7 givenname: Guifang surname: Wang fullname: Wang, Guifang organization: Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China – sequence: 8 givenname: Jingxiang surname: Zhong fullname: Zhong, Jingxiang organization: Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/25333784$$D View this record in MEDLINE/PubMed |
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| Notes | Conceived and designed the experiments: QJ JZ. Performed the experiments: BQ QJ LL. Analyzed the data: XG GH. Contributed reagents/materials/analysis tools: BQ QJ GW. Wrote the paper: BQ QJ YW. Competing Interests: The authors have declared that no competing interests exist. |
| OpenAccessLink | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4198253/ |
| PMID | 25333784 |
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| PQPubID | 1436336 |
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| PublicationCentury | 2000 |
| PublicationDate | 2014-10-15 |
| PublicationDateYYYYMMDD | 2014-10-15 |
| PublicationDate_xml | – month: 10 year: 2014 text: 2014-10-15 day: 15 |
| PublicationDecade | 2010 |
| PublicationPlace | United States |
| PublicationPlace_xml | – name: United States – name: San Francisco – name: San Francisco, USA |
| PublicationTitle | PloS one |
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| PublicationYear | 2014 |
| Publisher | Public Library of Science Public Library of Science (PLoS) |
| Publisher_xml | – name: Public Library of Science – name: Public Library of Science (PLoS) |
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| Snippet | We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress-induced apoptosis and senescence in human lens... Objectives We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress-induced apoptosis and senescence in... Objectives We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress–induced apoptosis and senescence in... Objectives We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress–induced apoptosis and senescence in... |
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| SubjectTerms | Aging Analysis Antioxidants Antioxidants (Nutrients) Apoptosis Apoptosis - drug effects Bax protein Bcl-2 protein bcl-2-Associated X Protein - metabolism Biology and Life Sciences Cataracts Cell Line, Transformed Cell lines Cellular Senescence - drug effects Cholecystokinin Diabetes Drugs, Chinese Herbal - pharmacology Enzymatic activity Enzyme activity Enzymes Epithelial cells Epithelial Cells - drug effects Epithelial Cells - metabolism Flow cytometry Galactosidase Glutathione Humans Hydrogen Hydrogen peroxide Hydrogen Peroxide - pharmacology Ischemia Lens, Crystalline - cytology Lenses Lycium - chemistry Lycium barbarum Malondialdehyde Medicine and Health Sciences Membrane potential Membrane Potential, Mitochondrial - drug effects Mitochondria Mitochondrial DNA Oxidative stress Oxidative Stress - drug effects Oxygen Polysaccharides Pretreatment Protective Agents - pharmacology Proteins Proto-Oncogene Proteins c-bcl-2 - metabolism Reactive oxygen species Reactive Oxygen Species - metabolism Rodents Saccharides Senescence Signal Transduction - drug effects Superoxide dismutase Superoxides β-Galactosidase |
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| Title | Lycium barbarum polysaccharides protect human lens epithelial cells against oxidative stress-induced apoptosis and senescence |
| URI | https://www.ncbi.nlm.nih.gov/pubmed/25333784 https://www.proquest.com/docview/1612297916 https://pubmed.ncbi.nlm.nih.gov/PMC4198253 https://doaj.org/article/76e2d79cbc334c50be67f937b8606aa5 http://dx.doi.org/10.1371/journal.pone.0110275 |
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