Suppression of tumor necrosis factor alpha-induced matrix metalloproteinase 9 production in human salivary gland acinar cells by cepharanthine occurs via down-regulation of nuclear factor kappaB: a possible therapeutic agent for preventing the destruction of the acinar structure in the salivary glands of Sjögren's syndrome patients.
OBJECTIVE
Our previous results suggested that suppression of tumor necrosis factor alpha (TNFalpha)-induced matrix metalloproteinase 9 (MMP-9) could prevent the destruction of acinar tissue in the salivary glands of patients with Sjögren's syndrome (SS). The present study was undertaken to investigate the effect of cepharanthine on the suppression of TNFalpha-induced MMP-9 production in NS-SV-AC, an SV40-immortalized normal human acinar cell clone.


METHODS
After pretreatment with or without cepharanthine, NS-SV-AC cells were treated with TNFalpha alone or with a combination of TNFalpha and cepharanthine. The expression of MMP-9 was then examined at the protein and messenger RNA levels. In addition, the effect of cepharanthine on the morphogenetic behavior of NS-SV-AC cells cultured on type IV collagen-coated dishes in the presence of TNFalpha was examined.


RESULTS
Although TNFalpha induced the production of MMP-9 in NS-SV-AC cells, this production was greatly suppressed when cells were pretreated with cepharanthine, followed by treatment with both TNFalpha and cepharanthine. In addition, cepharanthine suppressed the TNFalpha-stimulated NF-kappaB activity by partly preventing the degradation of IkappaBalpha protein in NS-SV-AC cells. When NS-SV-AC cells were seeded on type IV collagen-coated dishes in the presence of both TNFalpha and plasmin, type IV collagen interaction with the cells was lost and the cells entered apoptosis. However, pretreatment with cepharanthine restored the aberrant in vitro morphogenesis of the NS-SV-AC cells.


CONCLUSIONS
These results may indicate a molecular mechanism by which cepharanthine is able to protect against the destruction of the acinar structure in salivary glands from patients with SS.
Department of Oral and Maxillofacial Surgery 2, Tokushima University School of Dentistry, 3 Kuramoto-cho, Tokushima 770-8504, Japan. azumasa@dent.tokushima-u.ac.jp
Masayuki
M
Azuma
Keiko
K
Aota
Tetsuya
T
Tamatani
Katsumi
K
Motegi
Tsuyoshi
T
Yamashita
Yuki
Y
Ashida
Yoshio
Y
Hayashi
Mitsunobu
M
Sato
D000470:Alkaloids / Q000494:pharmacology*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D000906:Antibodies / Q000494:pharmacology; D044182:Benzylisoquinolines; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D024043:Collagen Type II / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D020672:I-kappa B Proteins; D020780:Matrix Metalloproteinase 9 / Q000235:genetics* / Q000378:metabolism; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000187:drug effects; D012333:RNA, Messenger / Q000032:analysis; D012469:Salivary Glands / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism*; D052117:Benzodioxoles
D000470:Alkaloids; D000894:Anti-Inflammatory Agents, Non-Steroidal; D000906:Antibodies; D044182:Benzylisoquinolines; D024043:Collagen Type II; D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D020780:Matrix Metalloproteinase 9; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; C000606204:NFKBIA protein, human; C006947:cepharanthine; D052117:Benzodioxoles