Neuroprotective Effects of Hesperidin, a Plant Flavanone, on Rotenone-Induced Oxidative Stress and Apoptosis in a Cellular Model for Parkinson's Disease
Identifieur interne : 001089 ( Pmc/Curation ); précédent : 001088; suivant : 001090Neuroprotective Effects of Hesperidin, a Plant Flavanone, on Rotenone-Induced Oxidative Stress and Apoptosis in a Cellular Model for Parkinson's Disease
Auteurs : Kuppusamy Tamilselvam [Inde] ; Nady Braidy [Australie] ; Thamilarasan Manivasagam [Inde] ; Musthafa Mohamed Essa [Oman] ; Nagarajan Rajendra Prasad [Inde] ; Subburayan Karthikeyan [Inde] ; Arokyasamy Justin Thenmozhi [Inde] ; Subash Selvaraju [Oman] ; Gilles J. Guillemin [Australie]Source :
- Oxidative Medicine and Cellular Longevity [ 1942-0900 ] ; 2013.
Abstract
Rotenone a widely used pesticide that inhibits mitochondrial complex I has been used to investigate the pathobiology of PD both
Url:
DOI: 10.1155/2013/102741
PubMed: 24205431
PubMed Central: 3800605
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Neuroprotective Effects of Hesperidin, a Plant Flavanone, on Rotenone-Induced Oxidative Stress and Apoptosis in a Cellular Model for Parkinson's Disease</title><author><name sortKey="Tamilselvam, Kuppusamy" sort="Tamilselvam, Kuppusamy" uniqKey="Tamilselvam K" first="Kuppusamy" last="Tamilselvam">Kuppusamy Tamilselvam</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Braidy, Nady" sort="Braidy, Nady" uniqKey="Braidy N" first="Nady" last="Braidy">Nady Braidy</name><affiliation wicri:level="1"><nlm:aff id="I2">Centre for Healthy Brain Ageing, School of Psychiatry, Faculty of Medicine, University of New SouthWales, Sydney 2031, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>Centre for Healthy Brain Ageing, School of Psychiatry, Faculty of Medicine, University of New SouthWales, Sydney 2031</wicri:regionArea></affiliation></author><author><name sortKey="Manivasagam, Thamilarasan" sort="Manivasagam, Thamilarasan" uniqKey="Manivasagam T" first="Thamilarasan" last="Manivasagam">Thamilarasan Manivasagam</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Essa, Musthafa Mohamed" sort="Essa, Musthafa Mohamed" uniqKey="Essa M" first="Musthafa Mohamed" last="Essa">Musthafa Mohamed Essa</name><affiliation wicri:level="1"><nlm:aff id="I3">Department of Food Science and Nutrition, College of Agriculture and Marine Sciences, Sultan Qaboos University, P.O. Box 50, Muscat 123, Oman</nlm:aff><country xml:lang="fr">Oman</country><wicri:regionArea>Department of Food Science and Nutrition, College of Agriculture and Marine Sciences, Sultan Qaboos University, P.O. Box 50, Muscat 123</wicri:regionArea></affiliation></author><author><name sortKey="Prasad, Nagarajan Rajendra" sort="Prasad, Nagarajan Rajendra" uniqKey="Prasad N" first="Nagarajan Rajendra" last="Prasad">Nagarajan Rajendra Prasad</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Karthikeyan, Subburayan" sort="Karthikeyan, Subburayan" uniqKey="Karthikeyan S" first="Subburayan" last="Karthikeyan">Subburayan Karthikeyan</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Thenmozhi, Arokyasamy Justin" sort="Thenmozhi, Arokyasamy Justin" uniqKey="Thenmozhi A" first="Arokyasamy Justin" last="Thenmozhi">Arokyasamy Justin Thenmozhi</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Selvaraju, Subash" sort="Selvaraju, Subash" uniqKey="Selvaraju S" first="Subash" last="Selvaraju">Subash Selvaraju</name><affiliation wicri:level="1"><nlm:aff id="I3">Department of Food Science and Nutrition, College of Agriculture and Marine Sciences, Sultan Qaboos University, P.O. Box 50, Muscat 123, Oman</nlm:aff><country xml:lang="fr">Oman</country><wicri:regionArea>Department of Food Science and Nutrition, College of Agriculture and Marine Sciences, Sultan Qaboos University, P.O. Box 50, Muscat 123</wicri:regionArea></affiliation></author><author><name sortKey="Guillemin, Gilles J" sort="Guillemin, Gilles J" uniqKey="Guillemin G" first="Gilles J." last="Guillemin">Gilles J. Guillemin</name><affiliation wicri:level="1"><nlm:aff id="I4">Neuropharmacology Group, MND and Neurodegenerative Diseases Research Centre, Australian School of Advanced Medicine, Macquarie University, Balaclava Road, North Ryde, Sydney, NSW 2109, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>Neuropharmacology Group, MND and Neurodegenerative Diseases Research Centre, Australian School of Advanced Medicine, Macquarie University, Balaclava Road, North Ryde, Sydney, NSW 2109</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">24205431</idno><idno type="pmc">3800605</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3800605</idno><idno type="RBID">PMC:3800605</idno><idno type="doi">10.1155/2013/102741</idno><date when="2013">2013</date><idno type="wicri:Area/Pmc/Corpus">001090</idno><idno type="wicri:Area/Pmc/Curation">001089</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Neuroprotective Effects of Hesperidin, a Plant Flavanone, on Rotenone-Induced Oxidative Stress and Apoptosis in a Cellular Model for Parkinson's Disease</title><author><name sortKey="Tamilselvam, Kuppusamy" sort="Tamilselvam, Kuppusamy" uniqKey="Tamilselvam K" first="Kuppusamy" last="Tamilselvam">Kuppusamy Tamilselvam</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Braidy, Nady" sort="Braidy, Nady" uniqKey="Braidy N" first="Nady" last="Braidy">Nady Braidy</name><affiliation wicri:level="1"><nlm:aff id="I2">Centre for Healthy Brain Ageing, School of Psychiatry, Faculty of Medicine, University of New SouthWales, Sydney 2031, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>Centre for Healthy Brain Ageing, School of Psychiatry, Faculty of Medicine, University of New SouthWales, Sydney 2031</wicri:regionArea></affiliation></author><author><name sortKey="Manivasagam, Thamilarasan" sort="Manivasagam, Thamilarasan" uniqKey="Manivasagam T" first="Thamilarasan" last="Manivasagam">Thamilarasan Manivasagam</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Essa, Musthafa Mohamed" sort="Essa, Musthafa Mohamed" uniqKey="Essa M" first="Musthafa Mohamed" last="Essa">Musthafa Mohamed Essa</name><affiliation wicri:level="1"><nlm:aff id="I3">Department of Food Science and Nutrition, College of Agriculture and Marine Sciences, Sultan Qaboos University, P.O. Box 50, Muscat 123, Oman</nlm:aff><country xml:lang="fr">Oman</country><wicri:regionArea>Department of Food Science and Nutrition, College of Agriculture and Marine Sciences, Sultan Qaboos University, P.O. Box 50, Muscat 123</wicri:regionArea></affiliation></author><author><name sortKey="Prasad, Nagarajan Rajendra" sort="Prasad, Nagarajan Rajendra" uniqKey="Prasad N" first="Nagarajan Rajendra" last="Prasad">Nagarajan Rajendra Prasad</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Karthikeyan, Subburayan" sort="Karthikeyan, Subburayan" uniqKey="Karthikeyan S" first="Subburayan" last="Karthikeyan">Subburayan Karthikeyan</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Thenmozhi, Arokyasamy Justin" sort="Thenmozhi, Arokyasamy Justin" uniqKey="Thenmozhi A" first="Arokyasamy Justin" last="Thenmozhi">Arokyasamy Justin Thenmozhi</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002</wicri:regionArea></affiliation></author><author><name sortKey="Selvaraju, Subash" sort="Selvaraju, Subash" uniqKey="Selvaraju S" first="Subash" last="Selvaraju">Subash Selvaraju</name><affiliation wicri:level="1"><nlm:aff id="I3">Department of Food Science and Nutrition, College of Agriculture and Marine Sciences, Sultan Qaboos University, P.O. Box 50, Muscat 123, Oman</nlm:aff><country xml:lang="fr">Oman</country><wicri:regionArea>Department of Food Science and Nutrition, College of Agriculture and Marine Sciences, Sultan Qaboos University, P.O. Box 50, Muscat 123</wicri:regionArea></affiliation></author><author><name sortKey="Guillemin, Gilles J" sort="Guillemin, Gilles J" uniqKey="Guillemin G" first="Gilles J." last="Guillemin">Gilles J. Guillemin</name><affiliation wicri:level="1"><nlm:aff id="I4">Neuropharmacology Group, MND and Neurodegenerative Diseases Research Centre, Australian School of Advanced Medicine, Macquarie University, Balaclava Road, North Ryde, Sydney, NSW 2109, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>Neuropharmacology Group, MND and Neurodegenerative Diseases Research Centre, Australian School of Advanced Medicine, Macquarie University, Balaclava Road, North Ryde, Sydney, NSW 2109</wicri:regionArea></affiliation></author></analytic><series><title level="j">Oxidative Medicine and Cellular Longevity</title><idno type="ISSN">1942-0900</idno><idno type="eISSN">1942-0994</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Rotenone a widely used pesticide that inhibits mitochondrial complex I has been used to investigate the pathobiology of PD both <italic>in vitro</italic> and <italic>in vivo</italic>. Studies have shown that the neurotoxicity of rotenone may be related to its ability to generate reactive oxygen species (ROS), leading to neuronal apoptosis. The current study was carried out to investigate the neuroprotective effects of hesperidin, a citrus fruit flavanol, against rotenone-induced apoptosis in human neuroblastoma SK-N-SH cells. We assessed cell death, mitochondrial membrane potential, ROS generation, ATP levels, thiobarbituric acid reactive substances, reduced glutathione (GSH) levels, and the activity of catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx) using well established assays. Apoptosis was determined in normal, rotenone, and hesperidin treated cells, by measuring the protein expression of cytochrome c (cyt c), caspases 3 and 9, Bax, and Bcl-2 using the standard western blotting technique. The apoptosis in rotenone-induced SK-N-SH cells was accompanied by the loss of mitochondrial membrane potential, increased ROS generation, the depletion of GSH, enhanced activities of enzymatic antioxidants, upregulation of Bax, cyt c, and caspases 3 and 9, and downregulation of Bcl-2, which were attenuated in the presence of hesperidin. Our data suggests that hesperidin exerts its neuroprotective effect against rotenone due to its antioxidant, maintenance of mitochondrial function, and antiapoptotic properties in a neuroblastoma cell line.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Dauer, W" uniqKey="Dauer W">W Dauer</name></author><author><name sortKey="Przedborski, S" uniqKey="Przedborski S">S Przedborski</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Anantharam, V" uniqKey="Anantharam V">V Anantharam</name></author><author><name sortKey="Lehrmann, E" uniqKey="Lehrmann E">E Lehrmann</name></author><author><name sortKey="Kanthasamy, A" uniqKey="Kanthasamy A">A Kanthasamy</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jenner, P" uniqKey="Jenner P">P Jenner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Przedborski, S" uniqKey="Przedborski S">S 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Oxid Med Cell Longev</journal-id><journal-id journal-id-type="iso-abbrev">Oxid Med Cell Longev</journal-id><journal-id journal-id-type="publisher-id">OXIMED</journal-id><journal-title-group><journal-title>Oxidative Medicine and Cellular Longevity</journal-title></journal-title-group><issn pub-type="ppub">1942-0900</issn><issn pub-type="epub">1942-0994</issn><publisher><publisher-name>Hindawi Publishing Corporation</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">24205431</article-id><article-id pub-id-type="pmc">3800605</article-id><article-id pub-id-type="doi">10.1155/2013/102741</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title>Neuroprotective Effects of Hesperidin, a Plant Flavanone, on Rotenone-Induced Oxidative Stress and Apoptosis in a Cellular Model for Parkinson's Disease</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Tamilselvam</surname><given-names>Kuppusamy</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">http://orcid.org/0000-0002-0497-5572</contrib-id><name><surname>Braidy</surname><given-names>Nady</given-names></name><xref ref-type="aff" rid="I2"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Manivasagam</surname><given-names>Thamilarasan</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Essa</surname><given-names>Musthafa Mohamed</given-names></name><xref ref-type="aff" rid="I3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Prasad</surname><given-names>Nagarajan Rajendra</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Karthikeyan</surname><given-names>Subburayan</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Thenmozhi</surname><given-names>Arokyasamy Justin</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Selvaraju</surname><given-names>Subash</given-names></name><xref ref-type="aff" rid="I3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Guillemin</surname><given-names>Gilles J.</given-names></name><xref ref-type="aff" rid="I4"><sup>4</sup></xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="I1"><sup>1</sup>Department of Biochemistry and Biotechnology, Faculty of Science, College Rd, Annamalai Nagar, Chidambaram, Tamil Nadu 608002, India</aff><aff id="I2"><sup>2</sup>Centre for Healthy Brain Ageing, School of Psychiatry, Faculty of Medicine, University of New SouthWales, Sydney 2031, Australia</aff><aff id="I3"><sup>3</sup>Department of Food Science and Nutrition, College of Agriculture and Marine Sciences, Sultan Qaboos University, P.O. Box 50, Muscat 123, Oman</aff><aff id="I4"><sup>4</sup>Neuropharmacology Group, MND and Neurodegenerative Diseases Research Centre, Australian School of Advanced Medicine, Macquarie University, Balaclava Road, North Ryde, Sydney, NSW 2109, Australia</aff><author-notes><corresp id="cor1">*Gilles J. Guillemin: <email>gilles.guillemin@mq.edu.au</email></corresp><fn fn-type="other"><p>Academic Editor: Tullia Maraldi</p></fn></author-notes><pub-date pub-type="ppub"><year>2013</year></pub-date><pub-date pub-type="epub"><day>24</day><month>9</month><year>2013</year></pub-date><volume>2013</volume><elocation-id>102741</elocation-id><history><date date-type="received"><day>8</day><month>7</month><year>2013</year></date><date date-type="accepted"><day>15</day><month>8</month><year>2013</year></date></history><permissions><copyright-statement>Copyright © 2013 Kuppusamy Tamilselvam et al.</copyright-statement><copyright-year>2013</copyright-year><license license-type="open-access"><license-p>This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><abstract><p>Rotenone a widely used pesticide that inhibits mitochondrial complex I has been used to investigate the pathobiology of PD both <italic>in vitro</italic> and <italic>in vivo</italic>. Studies have shown that the neurotoxicity of rotenone may be related to its ability to generate reactive oxygen species (ROS), leading to neuronal apoptosis. The current study was carried out to investigate the neuroprotective effects of hesperidin, a citrus fruit flavanol, against rotenone-induced apoptosis in human neuroblastoma SK-N-SH cells. We assessed cell death, mitochondrial membrane potential, ROS generation, ATP levels, thiobarbituric acid reactive substances, reduced glutathione (GSH) levels, and the activity of catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx) using well established assays. Apoptosis was determined in normal, rotenone, and hesperidin treated cells, by measuring the protein expression of cytochrome c (cyt c), caspases 3 and 9, Bax, and Bcl-2 using the standard western blotting technique. The apoptosis in rotenone-induced SK-N-SH cells was accompanied by the loss of mitochondrial membrane potential, increased ROS generation, the depletion of GSH, enhanced activities of enzymatic antioxidants, upregulation of Bax, cyt c, and caspases 3 and 9, and downregulation of Bcl-2, which were attenuated in the presence of hesperidin. Our data suggests that hesperidin exerts its neuroprotective effect against rotenone due to its antioxidant, maintenance of mitochondrial function, and antiapoptotic properties in a neuroblastoma cell line.</p></abstract></article-meta></front><floats-group><fig id="fig1" orientation="portrait" position="float"><label>Figure 1</label><caption><p>Effect of hesperidin on rotenone-induced reduction in cell proliferation in SK-N-SH neuroblastoma cells. (a) The dose-dependent effect of rotenone (0.5, 5, 50, 100, and 200 nM) on cell proliferation after 24 h. Values are presented as mean ± SD of four experiments in each group. 50% inhibition concentration value (IC<sub>50</sub>) was found to be 100 nM. ∗ indicates significance compared to nontreated cells. (b) The dose-dependent effect of hesperidin (2.5, 5, 10, 20, and 40 <italic>μ</italic>g) alone and against rotenone-induced changes on cell proliferation. Values are presented as mean ± SD of four experiments in each group. Treatment with hesperidin alone (blue column) (2.5, 5, 10, 20, and 40 <italic>μ</italic>g) did not affect cell proliferation. Hesperidin (2.5, 5, 10, and 20 <italic>μ</italic>g) pretreatment dose dependently enhanced cell proliferation against rotenone toxicity.</p></caption><graphic xlink:href="OXIMED2013-102741.001"></graphic></fig><fig id="fig2" orientation="portrait" position="float"><label>Figure 2</label><caption><p>Effect of hesperidin (20 <italic>μ</italic>g) on rotenone (100 nM)-induced oxidative and antioxidative indices. Rotenone treatment significantly increased and decreased the levels of TBARS and GSH, respectively, as compared to control cells, while hesperidin pretreatment significantly decreased and enhanced the levels of TBARS and GSH as compared to rotenone alone treated cells (Figures <xref ref-type="fig" rid="fig2">2(a)</xref> and <xref ref-type="fig" rid="fig2">2(b)</xref>). Values are presented as mean ± SD of four experiments in each group. *<italic>P</italic> < 0.05 compared to control, and <sup>#</sup><italic>P</italic> < 0.05 compared to rotenone group (DMRT). Rotenone treatment enhanced the activities of SOD, CAT, and GPx as compared to untreated cells, while hesperidin pretreatment significantly downregulated the activities of enzymatic antioxidants as compared to rotenone alone treated cells ((c), (d), and (e)). Values are given as mean ± SD of four experiments in each group. <sup>1</sup>Enzyme concentration required for 50% inhibition of nitroblue tetrazolium reduction in 1 min. <sup>2</sup>Micromoles of hydrogen peroxide consumed per minute. <sup>3</sup>Micrograms of glutathione consumed per minute.</p></caption><graphic xlink:href="OXIMED2013-102741.002"></graphic></fig><fig id="fig3" orientation="portrait" position="float"><label>Figure 3</label><caption><p>Effect of hesperidin on rotenone-induced ROS generation in SK-N-SH cells. (a) Microscopic images showing the preventive effect of hesperidin against rotenone-induced ROS generation by DCFDA staining. (b) Rotenone (100 nM) treatment significantly increased the levels of ROS as compared to control cells, while hesperidin (20 <italic>μ</italic>g) pretreatment significantly decreased the levels of ROS as compared to rotenone alone treated cells. Values are given as mean ± S.D. of four experiments in each group. *<italic>P</italic> < 0.05 compared to control, and <sup>#</sup><italic>P</italic> < 0.05 compared to rotenone group (DMRT).</p></caption><graphic xlink:href="OXIMED2013-102741.003"></graphic></fig><fig id="fig4" orientation="portrait" position="float"><label>Figure 4</label><caption><p>Measurement of intracellular ATP levels. Rotenone (100 nM) treatment significantly reduced the levels of ATP as compared to control cells, while hesperidin (20 <italic>μ</italic>g) pretreatment significantly enhanced the levels of ATP as compared to rotenone alone treated cells. Values are given as mean ± S.D. of four experiments in each group. *<italic>P</italic> < 0.05 compared to control, and <sup>#</sup><italic>P</italic> < 0.05 compared to rotenone group (DMRT).</p></caption><graphic xlink:href="OXIMED2013-102741.004"></graphic></fig><fig id="fig5" orientation="portrait" position="float"><label>Figure 5</label><caption><p>Alteration in mitochondrial membrane potential of control, hesperidin, and rotenone-treated SK-N-SH cells. Rotenone (100 nM) significantly decreased mitochondrial membrane potential, while hesperidin (20 <italic>μ</italic>g) pretreatment significantly increased MMP in rotenone-treated SK-N-SH cells ((a) and (b)). Values are given as mean ± S.D of four experiments in each group. *<italic>P</italic> < 0.05 compared to control, and <sup>#</sup><italic>P</italic> < 0.05 compared to rotenone groups (DMRT).</p></caption><graphic xlink:href="OXIMED2013-102741.005"></graphic></fig><fig id="fig6" orientation="portrait" position="float"><label>Figure 6</label><caption><p>Effect of hesperidin on rotenone-induced Bax and Bcl-2 expressions in SK-N-SH cells. Rotenone (100 nM) significantly enhanced the expression of Bax and diminished the expression of Bcl-2, while hesperidin (20 <italic>μ</italic>g) pretreatment significantly diminished the expression of Bax and elevated the expression of Bcl-2 in rotenone-treated SK-N-SH cells ((a) and (b)). Western blot data are quantified by using <italic>β</italic>-actin as an internal control, and the values are expressed as arbitrary units and given as mean ± SD of four experiments in each group. *<italic>P</italic> < 0.05 compared to control, and <sup>#</sup><italic>P</italic> < 0.05 compared to rotenone alone treated group.</p></caption><graphic xlink:href="OXIMED2013-102741.006"></graphic></fig><fig id="fig7" orientation="portrait" position="float"><label>Figure 7</label><caption><p>Effect of hesperidin on rotenone-induced cyt c and caspases 3 and 9 expressions in SK-N-SH cells. Rotenone (100 nM) significantly enhanced the expressions of cyt c, caspases 3 and 9, while hesperidin (20 <italic>μ</italic>g) pretreatment significantly diminished the expressions of cyt c, caspases 3 and 9 in rotenone-treated SK-N-SH cells ((a), (b), and (c)). Western blot data are quantified by using <italic>β</italic>-actin as an internal control, and the values are expressed as arbitrary units and given as mean ± SD of four experiments in each group. *<italic>P</italic> < 0.05 compared to control, <sup>#</sup><italic>P</italic> < 0.05 compared to rotenone groups.</p></caption><graphic xlink:href="OXIMED2013-102741.007"></graphic></fig></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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