Cisplatin regulates Sertoli cell expression of transferrin and interleukins.
It is well known that exposure to cis-diaminedichloroplatinum (CDDP) results in impairment of spermatogenesis; however, little is known about the signal mechanisms involved in CDDP regulation of Sertoli cell (SC) function. This study was designed to evaluate how CDDP regulates SC signal molecules and mechanisms. Purified rat SC was cultured under serum-free conditions and treated with CDDP (10 ng/ml) at various time points. Western blot analysis was used to determine the activation of extracellular signal-related kinases 1 and 2 (ERK1/2), p38 mitogen-activated protein kinase (MAPK), cJun-N-terminal kinase (JNK), cyclooxygenase (COX)-1 and COX-2, inducible and endothelial nitric oxide synthase (iNOS and eNOS). The levels of transferrin (TF) and prostaglandin (PG)E2, PGF2alpha, PGD2, carbaprostacyclin (cPGI2 analog) in culture medium were quantified by ELISA. Nitrite (NO2(-)) and nitrate (NO3(-)) in culture medium were also quantified by Griess assay. Interleukin (IL)-1beta and IL-6 mRNAs were measured by quantitative real-time PCR (QRT-PCR) analysis. CDDP activated the phosphorylation of ERK1/2 (p-ERK1/2) in the early phase (within 5 min) and the level of transferrin (TF) fell significantly. In addition, CDDP significantly increased the level of COX-2, PGs, and ILs in the late phase (within 24h). When ERK activity inhibitor (PD98059, 10 microM) or COX-2 activity inhibitor (NS-398, 10 microM) was used, CDDP reduction of TF and induction of PG and IL expression were prevented, suggesting that the detrimental effects on spermatogenesis through the impairment of SC induced by CDDP are mediated by the activation of ERK1/2 and COX-2 pathways in SC.
Yamaguchi
Kohei
K
Division of Urology, Department of Organs Therapeutics, Faculty of Medicine, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-Cho, Chuo-Ku, Kobe 650-0017, Japan.
Ishikawa
Tomomoto
T
Kondo
Yutaka
Y
Fujisawa
Masato
M
D000818:Animals; D002478:Cells, Cultured; D002945:Cisplatin / Q000494:pharmacology*; D051546:Cyclooxygenase 2 / Q000378:metabolism; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism*; D008297:Male; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D011453:Prostaglandins / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D014168:Transferrin / Q000378:metabolism*
D016861:Cyclooxygenase Inhibitors; D053583:Interleukin-1beta; D015850:Interleukin-6; D011453:Prostaglandins; D014168:Transferrin; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D051546:Cyclooxygenase 2; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D002945:Cisplatin