Protective effect of Hexane and Ethanol extract of piper longum L. on gentamicin-induced hair cell loss in neonatal cultures

Mukesh Kumar Yadav, June Choi, Jae-Jun Song

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Objectives. Gentamicin (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. Methods. The 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. Results. SPL 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. Conclusion. An 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.

Original languageEnglish
Pages (from-to)13-18
Number of pages6
JournalClinical and Experimental Otorhinolaryngology
Volume7
Issue number1
DOIs
Publication statusPublished - 2014 Feb 27

Fingerprint

Piper
Alopecia
Hexanes
Gentamicins
Ethanol
Cochlea
Apoptosis
Staining and Labeling
Stereocilia
Pacific Islands
Phalloidine
Spices
DNA Nucleotidylexotransferase
Aminoglycosides
Traditional Medicine
Inner Ear
Myosins
Transferases
Hair
Free Radicals

Keywords

  • Antioxidant
  • Apoptosis
  • Autotoxicity
  • Gentamicin
  • Inner hair cell
  • Outer hair cell
  • Piper longum

ASJC Scopus subject areas

  • Surgery
  • Otorhinolaryngology

Cite this

@article{b3a09d8b070545bc97b5994f4963534d,
title = "Protective effect of Hexane and Ethanol extract of piper longum L. on gentamicin-induced hair cell loss in neonatal cultures",
abstract = "Objectives. Gentamicin (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. Methods. The 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. Results. SPL 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. Conclusion. An 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.",
keywords = "Antioxidant, Apoptosis, Autotoxicity, Gentamicin, Inner hair cell, Outer hair cell, Piper longum",
author = "Yadav, {Mukesh Kumar} and June Choi and Jae-Jun Song",
year = "2014",
month = "2",
day = "27",
doi = "10.3342/ceo.2014.7.1.13",
language = "English",
volume = "7",
pages = "13--18",
journal = "Clinical and Experimental Otorhinolaryngology",
issn = "1976-8710",
publisher = "Korean Society of Otolaryngology",
number = "1",

}

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T1 - Protective effect of Hexane and Ethanol extract of piper longum L. on gentamicin-induced hair cell loss in neonatal cultures

AU - Yadav, Mukesh Kumar

AU - Choi, June

AU - Song, Jae-Jun

PY - 2014/2/27

Y1 - 2014/2/27

N2 - Objectives. Gentamicin (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. Methods. The 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. Results. SPL 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. Conclusion. An 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.

AB - Objectives. Gentamicin (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. Methods. The 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. Results. SPL 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. Conclusion. An 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.

KW - Antioxidant

KW - Apoptosis

KW - Autotoxicity

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KW - Inner hair cell

KW - Outer hair cell

KW - Piper longum

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DO - 10.3342/ceo.2014.7.1.13

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