Protective Effect of Hexane and Ethanol Extract of L. on Gentamicin-Induced Hair Cell Loss in Neonatal Cultures
ObjectivesGentamicin (GM) is a commonly used aminoglycoside antibiotic that generates free oxygen radicals within the inner ear, which can cause vestibulo-cochlear toxicity and permanent damage to the sensory hair cells and neurons. Piper longum L. (PL) is a well-known spice and traditional medicine...
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| Format: | Article |
| Language: | English |
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Korean Society of Otorhinolaryngology-Head and Neck Surgery
2014-03-01
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| Series: | Clinical and Experimental Otorhinolaryngology |
| Subjects: | |
| Online Access: | http://www.e-ceo.org/upload/pdf/ceo-7-13.pdf |
| _version_ | 1828205143442587648 |
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| author | Mukesh Kumar Yadav June Choi Jae-Jun Song |
| author_facet | Mukesh Kumar Yadav June Choi Jae-Jun Song |
| author_sort | Mukesh Kumar Yadav |
| collection | DOAJ |
| description | ObjectivesGentamicin (GM) is a commonly used aminoglycoside antibiotic that generates free oxygen radicals within the inner ear, which can cause vestibulo-cochlear toxicity and permanent damage to the sensory hair cells and neurons. Piper longum L. (PL) is a well-known spice and traditional medicine in Asia and Pacific islands, which has been reported to exhibit a wide spectrum of activity, including antioxidant activity. In this study, we evaluated the effect of hexane:ethanol (2:8) PL extract (subfraction of PL [SPL] extract) on GM-induced hair cell loss in basal, middle and apical regions in a neonatal cochlea cultures.MethodsThe protective effects of SPL extract were measured by phalloidin staining of cultures from postnatal day 2-3 mice with GM-induced hair cell loss. The anti-apoptosis activity of SPL extract was measured using double labeling by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and myosin-7a staining. The radical-scavenging activity of SPL extract was assessed using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay.ResultsSPL extract at a concentration of 1 µg/mL significantly inhibited GM-induced hair cell loss at basal and middle region of cochlea, while 5 µg/mL was effective against apical region hair cell loss. The protective effect of SPL extract was concentration dependent and hair cells retained their stereocilia in explants treated with SPL extract prior to treatment with 0.3 mM GM. SPL extract decreased GM-induced apoptosis of hair cells as assessed by TUNEL staining. The outer hair and inner hair counts were not decreased in SPL extract treated groups in compare to GM treated explants. Additionally, SPL extract showed concentration dependent radical scavenging activity in a DPPH assay.ConclusionAn anti-apoptosis effect and potent radical scavenger activity of SPL extract protects from GM-induced hair cell loss at basal, middle and apical regions in neonatal cochlea cultures. |
| first_indexed | 2024-04-12T12:34:53Z |
| format | Article |
| id | doaj.art-1211bda70b4745658d846adab4952786 |
| institution | Directory Open Access Journal |
| issn | 1976-8710 2005-0720 |
| language | English |
| last_indexed | 2024-04-12T12:34:53Z |
| publishDate | 2014-03-01 |
| publisher | Korean Society of Otorhinolaryngology-Head and Neck Surgery |
| record_format | Article |
| series | Clinical and Experimental Otorhinolaryngology |
| spelling | doaj.art-1211bda70b4745658d846adab49527862022-12-22T03:32:55ZengKorean Society of Otorhinolaryngology-Head and Neck SurgeryClinical and Experimental Otorhinolaryngology1976-87102005-07202014-03-0171131810.3342/ceo.2014.7.1.13286Protective Effect of Hexane and Ethanol Extract of L. on Gentamicin-Induced Hair Cell Loss in Neonatal CulturesMukesh Kumar Yadav0June Choi1Jae-Jun Song2Department of Otorhinolaryngology-Head and Neck Surgery, Dongguk University Ilsan Hospital, Goyang, Korea.Department of Otorhinolaryngology-Head and Neck Surgery, Korea University College of Medicine, Seoul, Korea.Department of Otorhinolaryngology-Head and Neck Surgery, Dongguk University Ilsan Hospital, Goyang, Korea.ObjectivesGentamicin (GM) is a commonly used aminoglycoside antibiotic that generates free oxygen radicals within the inner ear, which can cause vestibulo-cochlear toxicity and permanent damage to the sensory hair cells and neurons. Piper longum L. (PL) is a well-known spice and traditional medicine in Asia and Pacific islands, which has been reported to exhibit a wide spectrum of activity, including antioxidant activity. In this study, we evaluated the effect of hexane:ethanol (2:8) PL extract (subfraction of PL [SPL] extract) on GM-induced hair cell loss in basal, middle and apical regions in a neonatal cochlea cultures.MethodsThe protective effects of SPL extract were measured by phalloidin staining of cultures from postnatal day 2-3 mice with GM-induced hair cell loss. The anti-apoptosis activity of SPL extract was measured using double labeling by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and myosin-7a staining. The radical-scavenging activity of SPL extract was assessed using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay.ResultsSPL extract at a concentration of 1 µg/mL significantly inhibited GM-induced hair cell loss at basal and middle region of cochlea, while 5 µg/mL was effective against apical region hair cell loss. The protective effect of SPL extract was concentration dependent and hair cells retained their stereocilia in explants treated with SPL extract prior to treatment with 0.3 mM GM. SPL extract decreased GM-induced apoptosis of hair cells as assessed by TUNEL staining. The outer hair and inner hair counts were not decreased in SPL extract treated groups in compare to GM treated explants. Additionally, SPL extract showed concentration dependent radical scavenging activity in a DPPH assay.ConclusionAn anti-apoptosis effect and potent radical scavenger activity of SPL extract protects from GM-induced hair cell loss at basal, middle and apical regions in neonatal cochlea cultures.http://www.e-ceo.org/upload/pdf/ceo-7-13.pdfGentamicinAutotoxicityInner hair cellOuter hair cellPiper longumApoptosisAntioxidant |
| spellingShingle | Mukesh Kumar Yadav June Choi Jae-Jun Song Protective Effect of Hexane and Ethanol Extract of L. on Gentamicin-Induced Hair Cell Loss in Neonatal Cultures Clinical and Experimental Otorhinolaryngology Gentamicin Autotoxicity Inner hair cell Outer hair cell Piper longum Apoptosis Antioxidant |
| title | Protective Effect of Hexane and Ethanol Extract of L. on Gentamicin-Induced Hair Cell Loss in Neonatal Cultures |
| title_full | Protective Effect of Hexane and Ethanol Extract of L. on Gentamicin-Induced Hair Cell Loss in Neonatal Cultures |
| title_fullStr | Protective Effect of Hexane and Ethanol Extract of L. on Gentamicin-Induced Hair Cell Loss in Neonatal Cultures |
| title_full_unstemmed | Protective Effect of Hexane and Ethanol Extract of L. on Gentamicin-Induced Hair Cell Loss in Neonatal Cultures |
| title_short | Protective Effect of Hexane and Ethanol Extract of L. on Gentamicin-Induced Hair Cell Loss in Neonatal Cultures |
| title_sort | protective effect of hexane and ethanol extract of l on gentamicin induced hair cell loss in neonatal cultures |
| topic | Gentamicin Autotoxicity Inner hair cell Outer hair cell Piper longum Apoptosis Antioxidant |
| url | http://www.e-ceo.org/upload/pdf/ceo-7-13.pdf |
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