Adrenomedullin attenuates interleukin-1β-induced inflammation and apoptosis in rat Leydig cells via inhibition of NF-κB signaling pathway.
The aim of this paper is to investigate the protective effects of adrenomedullin (ADM) on interleukin-1β (IL-1β)-induced inflammation and apoptosis in rat Leydig cells and its underlying molecular mechanisms. Leydig cells were isolated from adult Sprague-Dawley rats. The cell culture was established by adding ADM 2h prior to 24h treatment with IL-1β-induced cytotoxicity. We detected cell viability and concentrations of testosterone, reactive oxygen species (ROS), malondialdehyde (MDA), and reduced glutathione (GSH). Gene expression levels were measured for inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2). Concentrations were detected for nitric oxide (NO) and prostaglandin E2 (PGE2). Apoptosis was assessed using terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Levels of gene expression and protein were detected for Bcl-2, Bax, caspase-3, and poly adenosine diphosphate-ribose polymerase (PARP). Protein levels were measured for nuclear factor kappa B (NF-κB) p65 and IκBα. ADM reduced IL-1β-induced cytotoxicity. ADM pretreatment significantly increased testosterone concentrations and decreased ROS, MDA, and GSH concentrations. ADM pretreatment inhibited IL-1β-induced inflammation in Leydig cells by decreasing the gene expression levels of iNOS and COX-2, as well as the concentrations of NO and PGE2. ADM pretreatment further decreased the number of TUNEL-positive stained Leydig cells, as confirmed by the increase in gene expression and protein levels of Bcl-2 and the decrease of Bax, caspase-3, and PARP levels. Moreover, ADM pretreatment inhibited NF-κB p65 phosphorylation and IκBα phosphorylation and degradation. ADM has potential anti-inflammatory and anti-apoptotic properties in IL-1β-induced rat Leydig cells, which might be related to NF-κB signaling pathway.
Department of Urology, Renmin Hospital of Wuhan University, No. 238 Liberation Road, Wuhan 430060, Hubei Province, China.
Wei
W
Hu
Department of Orthopedics, Renmin Hospital of Wuhan University, No. 238 Liberation Road, Wuhan 430060, Hubei Province, China. Electronic address: zhoupanghu@126.com.
Pang-hu
PH
Zhou
Department of Urology, Renmin Hospital of Wuhan University, No. 238 Liberation Road, Wuhan 430060, Hubei Province, China.
Ting
T
Rao
Department of Urology, Renmin Hospital of Wuhan University, No. 238 Liberation Road, Wuhan 430060, Hubei Province, China.
Xiao-bin
XB
Zhang
Department of Urology, Renmin Hospital of Wuhan University, No. 238 Liberation Road, Wuhan 430060, Hubei Province, China.
Wei
W
Wang
Department of Urology, Renmin Hospital of Wuhan University, No. 238 Liberation Road, Wuhan 430060, Hubei Province, China.
Li-jun
LJ
Zhang
D053607:Adrenomedullin / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D007249:Inflammation / Q000276:immunology / Q000517:prevention & control*; D053583:Interleukin-1beta / Q000276:immunology*; D007985:Leydig Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D016328:NF-kappa B / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects*; D013329:Structure-Activity Relationship
D053583:Interleukin-1beta; D016328:NF-kappa B; D053607:Adrenomedullin
Adrenomedullin; Apoptosis; Inflammation; Interleukin-1β; Leydig cell; NF-κB