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Down-regulation of iNOS and TNF-α expression by kaempferol via NF-κB inactivation in aged rat gingival tissues

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Abstract

The primary objective of this study was to evaluate the ability and mechanism of action of kaempferol, which is contained in extracts from Nelumbo nucifera, a well-known Oriental herb used in traditional medicine, with regard to the inhibition of iNOS and TNF-α expression in aged rat gingival tissues. We conducted an investigation into the age-related effects of kaempferol on reactive oxygen species (ROS) and GSH oxidative status in samples of aged gingival tissues. Western blotting was conducted in order to determine the expression of iNOS, TNF-α, p38 MAPK, NIK/IKK, p65 and IκBα in the sample tissues. Electrophoretic mobility shift assays (EMSA) were conducted in an effort to characterize the binding activities of NF-κB transcription factors in the aged rat gingival nuclear extracts. Our results indicate that kaempferol reduced ROS levels and augmented GSH levels in a dose-dependent manner in the aged gingival tissues. Kaempferol was shown to effect a significant reduction in iNOS and TNF-α protein levels, as compared to control gingival tissue samples. The results of Western blot analysis revealed that kaempferol treatment effected the reduction of iNOS and TNF-α expression, decreased nuclear p65 and increased cytosolic p65, down-regulation of Erk, p38, JNK and NIK/IKK expression. The EMSA results also indicated that kaempferol, when administered to the rat tissues, attenuated the NF-κB nuclear binding activity. Kaempferol may inhibit ROS generation via the inhibition of iNOS and TNF-α expression in aged gingival tissues, via the modulation of the NF-κB and mitogen-activated protein kinase (MAPK) pathways.

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Abbreviations

DCF:

2,7-dichlorofluorescein

DTNB:

5,5-dithiobis-2-nitrobenzoic acid

EMSA:

Electrophoretic mobility shift assays

H2DCFDA:

2,7-dichlorofluorescein diacetate

MAPK:

mitogen-activated protein kinase

ROS:

reactive oxygen species

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Acknowledgements

We are thankful to the Aging Tissue Bank (R21-2000-000-00021-0) funded by Korea Science and Engineering Foundation for supplying aged tissue.

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Authors and Affiliations

  1. Department of Oral Pathology, School of Dentistry, Pusan National University, Busan, 602-739, Korea

    Hyung Keun Kim & Hae Ryoun Park

  2. Department of Pediatric Dentistry, School of Dentistry, Pusan National University, Busan, 602-739, Korea

    Tae Sung Chung

  3. Department of Oral Microbiology, School of Dentistry and Research Institute for Oral Biotechnology, Pusan National University, Ami-dong, Seo-gu, Busan, 602-739, Korea

    Jin Chung

  4. Department of Pharmacy, College of Pharmacy, Pusan National University, Busan, 602-739, Korea

    Jun Sik Lee & Hae Young Chung

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  1. Hyung Keun Kim
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  2. Hae Ryoun Park
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  3. Jun Sik Lee
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  4. Tae Sung Chung
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  5. Hae Young Chung
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Correspondence to Jin Chung.

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Kim, H.K., Park, H.R., Lee, J.S. et al. Down-regulation of iNOS and TNF-α expression by kaempferol via NF-κB inactivation in aged rat gingival tissues. Biogerontology 8, 399–408 (2007). https://doi.org/10.1007/s10522-007-9083-9

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  • DOI: https://doi.org/10.1007/s10522-007-9083-9

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