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Immunohistochemical Detection of Apoptotic Proteins, p53/Bax and JNK/FasL Cascade, in the Lung of Rats Exposed to Cigarette Smoke

  • Organ Toxicity and Mechanisms
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Abstract

Lung disease is the leading and second-leading cause of death in women and men in Taiwan, respectively. Epidemiological studies conducted in Taiwan have shown that cigarette smoking is the principal risk factor of lung disease, but little is known about the association between apoptosis and cigarette smoke (CS)-induced lung pathogenesis. We designed an animal exposure system to study signal proteins involved in the process of apoptosis induced by smoking in rat terminal bronchiole. Rats were exposed to CS in doses of 5, 10, and 15 cigarettes, respectively, and the exposure lasted for 30 min, twice a day, 6 days a week for 1 month. Following which the rats were sacrificed and the lung tissues were analyzed by histopathological methods. The terminal bronchioles revealed mild to severe inflammation according to the doses of CS and marked lipid peroxidation, lymphocyte infiltration, congestion, and epithelial emphysema of alveolar spaces were also noted. Using an in situ cell death detection kit (TA300), the association of CS with apoptosis was determined in a concentration-dependent manner. Immunohistochemical evaluation showed that CS treatment produced an increase in the cellular levels of Bax, t-Bid, cleaved caspase-3, phospho-p53, phospho-JNK, and FasL but a decline in Bcl-2 and Mcl-1 (p<0.001 for all) in rat terminal bronchioles. The results provided evidences suggesting that exposure to CS not only induced apoptosis, but also involved p53/Bax and JNK/FasL cascade pathway.

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Abbreviations

CS:

Cigarette smoke

Bax:

Bcl-2 associated X protein

t-Bid:

Truncated Bid

Bcl-2:

B cell lymphoma-2

MAPK:

Mitogen-activated protein kinases family

JNK:

c-Jun N-terminal kinase

Fas-L:

Fas ligand

DAB:

Diaminobenzidine

iNOS:

Inducible NO synthase

NOx:

Nitrogen oxides

HPF:

High-power-field

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Acknowledgment

This work was supported by a grant from the National Science Council (NSC 94-2320-B040-039), Taiwan.

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Correspondence to Chau-Jong Wang.

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Wu, CH., Lin, HH., Yan, FP. et al. Immunohistochemical Detection of Apoptotic Proteins, p53/Bax and JNK/FasL Cascade, in the Lung of Rats Exposed to Cigarette Smoke. Arch Toxicol 80, 328–336 (2006). https://doi.org/10.1007/s00204-005-0050-4

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  • DOI: https://doi.org/10.1007/s00204-005-0050-4

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